Please note that r=1 and it's not being included.

I see you're starting with the unit circle, but have failed to mention that s=r*theta where r=1.

https://i.imx.to/i/2026/11/09/78gx84.jpg

social identity theory

we care more about the group than policy

would've been horrible

this is so important to remember

this is kinda sad

even effects tasks

we like our group

we kinda need those less interested or educated so to speak

others effect us more than we know/ we tend to be very multifaceted

interesting way to view parties

even impacts where we live

that's a very large jump and increase

so true

we're group based creatures

shows us groups matter a lot to us as humans

https://i.imx.to/i/2026/11/09/78gx83.jpg

This is at odds with techno-utopian visions of urban ag like the "vertical farming" put forward by Despommier and others https://www.nytimes.com/2025/02/18/science/dickson-despommier-dead.html

Curious about this. Is the aim inclusivity/participation?

Those that have the time and means

This makes sense, but I don't think it excludes regional scale (like the Costa Rican GI maps) or transect maps (like we saw in Sharifi 2016, 9)

important to acknowledge the latent history of colonialism in the term "exploration"

Are they encouraging an equivalence of map and territory? Probably not...maybe more like using mapping as provocation to re-orient flows of power.

aspirational

https://i.imx.to/i/2026/11/09/78gx82.jpg

https://i.imx.to/i/2026/11/09/78gx81.jpg

https://i.imx.to/i/2026/11/09/78gx7z.jpg

The results reported in this paper may be biased by the reuse of the same control samples for comparing multiple conditions (e.g. RES and SUS vs. CON in Fig. 1e,f). This issue--identified by Karl Pearson in 1897!--is explained in a slightly different context in a recent preprint by Nicol et al.

Comparisons of differential expression between groups should be performed by using an appropriate statistical model with an interaction term, as I discussed here.

https://i.imx.to/i/2026/11/09/78gx7y.jpg

https://i.imx.to/i/2026/11/09/78gx7x.jpg

Socialism with Chinese characteristics is the official ideology of the Chinese Communist Party(CCP) conceived by Deng Xiaoping. It maintains the foundational Socialist and Marxist principles but combines it with Capitalistic characteristics like free markets. It simply means adapting socialist/Marxist principles, which were sufficient for their time, and molding them into an ideology that is perfect for present-day China while considering its history and economic infrastructure.

This is in realization that bringing about this change, particularly legislature-wise, will be tedious, perhaps more than expected. Hilariously Robinson uses the pulling down of pants metaphorically as a commitment to the struggle to come, which is like an episode of constipation.

Chapter 39's hostage of world leaders at the World Economic Forum who come out of the experience with an unchanged mind is evidence of this. It will take time for key stakeholders' minds to change.

Eventually, terrorism seems to be what changes minds for Robinson's utopia.

https://i.imx.to/i/2026/11/09/78gx7w.jpg

I thought f(x)=cf(a(x+b))+d would be in here.

https://i.imx.to/i/2026/11/09/78gx7v.jpg

https://i.imx.to/i/2026/11/09/78gx7u.jpg

https://ipfs.indy0.net/ipfs/bafybeiarpglu7do76oa4lxuckpyweww5rrjwlmlbtc5tzpvhlegvyt4yt4/p/index.html

galls law

If an experiment produces a clear result but a later replication fails to reproduce it, how should researchers decide whether the original finding was unreliable or whether the difference came from the conditions of the replication?

x

An empty column might mean that the research is missing, but couldn't it also mean our search strategy failed to find the relevant studies? I think we should be careful about claiming a research gap based only on the papers we selected.

I noticed this when searching for my assignment. The same keywords gave very different numbers of results in ACM Digital Library and Google Scholar. It made me realise that the number of hits depends on how the database searches and indexes papers, not necessarily how much relevant research exists.

A paper investigating whose opinions matter when it comes to policy change found that policy makers are better at responding to advantaged groups than to disadvantaged ones.

Perhaps this is why one of the characters expresses a disillusionment with the thought a few government legislators, prone to "corruption[and] stupidity," get to have such consequential influence over the economy.

The other emphasizes the rule of law and insists to "make it stick", that is, to make it work, rather than dismissing it, or abandoning it altogether.

In Stanley's novel, this disillusionment with the system may embellish anarchy but that wouldn't be sustainable, at least not as far as The Ministry's goals of climate reform are concerned. What instead needs to happen is for the legislators to be less corrupt, less stupid. Given that the law mirrors the character of the legislators, perhaps the resultant policies will mirror a decline in "corruption" and "stupidity" in government legislators, and consequentially, climate reform will be a salient agendum.

actor model

• N supabase
• Variación N según tareas completadas
• Check manipulación
• Criterios Netquest

What will be the easiest-to-remember "brand" of academic-ready toy Earth System models we couldn't have dreamed of a year or two ago? Just wow

https://www.reddit.com/r/typewriters/comments/1xrobt6/script_typefaces_what_ive_learnt_in_tracking_them/

Broad colloquial advice on finding script machines. Fairly facile and not very nuanced, but broadly rings as true.

Zhiming Kuang matrix of linear response functions of a deep convecting model at RCE equilibrium

Advanced Typing - Shortcuts (1943)

Advanced Typing: Shortcuts. 16 mm. Vol. MN-1512c. United States Navy Training Film, 1943. https://www.youtube.com/watch?v=JUJfCfqgsX0.

Correct typing posture: fingers curved<br /> arms sloping up<br /> light fast strokes<br /> steady rhythm

fast continuous motion of return lever<br /> using backing sheet (aka temping sheet ??)

Shortcuts:<br /> mise en place for office supplies (she doesn't use this phrasing though) - greater efficiency - cuts down on searching

tabulators can be helpful. There are two types:<br /> - automatic - handset

Use tabs for paragraphs, dating letters, columns of numbers, and letter closings.

To clear all tab stops, put carriage to left, hold tab clear key and move the carriage across. (Usually applies to Royal, Remington, Underwood, and Electromatic).

On LC Smith and Woodstock machines sometimes have a tab clear lever on the back.

Decimal tabulator keys help to align a variety of numbers around a decimal point. 09:43

Always have a few tabs set to prevent a flying carriage which can be hard on the machine.

When using carbon sheets which are slightly longer than the paper size, cut off a small triangle at the top left hand side. This makes it easier for one to separate the carbons from the copies by holding the top left with one hand and pulling the carbons out from the bottom of the stack.

To align multiple sheets of paper for carbon copies, use a folded sheet at the top to taco the pages into the machine. Remove the folded sheet once the carbon pack is rolled forward.

Paper bail rollers should be set to split the pages into thirds (for two rollers).

Remington noiseless machines have a pressure indicator on the front of the machine (usually above the keyboard) which can be used when using thick carbon packs that may cause the ribbon guide to stick or bind.

Only erase when the carriage is fully left or right to prevent eraser crumbs from falling into the machine. 15:26

Use a soft eraser on carbon copies. Use and insert slips of paper behind the carbons and allowing them to stick out the sides, erasing from back sheet to front so as not to allow the eraser to mark your carbon copies. For the front sheet, use a shield and ink eraser and erase with a horizontal motion. After erasing, easily pull out the inserted sheets.

When typing a correction, tap the key lightly two or three times rather than hard once.

When in a rush and it's necessary to add a word (on double spacing), underline the last letter of the prior word and type a slash (/). Then move the typing line up and type the insertion above the prior line. This creates an "arrow" of sorts for the inserted word.

Details for inserting extra letters in misspelled words using half-spacing machines. (Underwoods and Electromatics don't have this function.)

Light pencil marks at the bottom of the sheet can help to indicate the coming bottom of the sheet.

Putting up the card holders (fingers) on Underwoods and Royals. They help to hold the card and improve print quality and reduce noise.

Card holders can cause markings on carbon packs if they're not lowered.

Trick for quickly writing postcards in succession: Disengage the ratchet using the platen spring release (or variable platen switch) Type the address on the front of the card. When done give the platen a quick practiced spin. The postcard with "jump" up and stop at the paper table and be in position for rolling in the opposite direction to write the message on the back of the card! When done a faster spin of the platen will shoot the card over the back of the typewriter where it can land in a box to collect all the postcards which were written in such a manner. <br /> timestamp 23:22

Time saving methods for addressing envelopes:

• Front seat principle. Insert the envelope in the usual way and type out the address. When done, turn the envelope down through the machine with the right hand. With the left hand, place the next envelope between the top of the first envelope and the front of the platen. Feed the first envelope back through the machine (in reverse) and the second will be rolled in to place for typing. Continue in this fashion until finished. All the finished envelopes will stack up in the back at the paper table.

• Chain feeding. The first envelope is inserted and rolled partway into the machine. A second envelope is inserted between the platen and the second envelope (behind the platen). Turn the first envelope to the writing line and type the address. Take out the first envelope and insert the next the same way as before.

• Uses paper bail. Do the first envelope in the usual way. Spin it out of the machine up and behind the paper bail into a box behind the typewriter.

For quickly doing labels or small cards: (27:29) Create a small zig-zag fold into a piece of paper to create a pocket slot which can be scotch taped on either side. This template paper can then be inserted so that the pocket is visible above the writing line, but the paper below it is still in the platen. The label or card can be placed into the pocket and the platen reversed to feed the label or card in backwards to the desired typing line. Using a v-groove or hole in the typing line can create a pencil line to serve as a guide for inserting many labels at the same place so that the typing lines up between labels.

Some offices had special platens for holding cards like this.

Pockets like this can also be used to hold the page to add additional lines at the bottom of pages. Deeper pockets may need to be used for doing this with carbon packs whose carbons are longer than the pages.

Alternately one can do something similar by creating a inverted u-shaped set of slits into an index card. to hold such labels.

When in the midst of a page and needing to do another piece urgently, roll back the letter until about 2 inches from the top, and then place in the new page and one between each of the carbons. Then roll forward to do the short message as necessary. Turn back to the insertion position to remove the copies and then continue with the first letter where you left off.

For drawing horizontal lines on typewriter paper, push the carriage to the extreme left and place the pencil or pen at the edge of the card guide and the scale. Then move the carriage to the right to effect the line. For vertical lines, put the carriage at the desired space and place the pencil at the card guide and scale and move the platen up/down as necessary.

ethos/citation

ethos/ citing

communities

Thesis stated in the end of the paragraph. Allows a momentum of build up for the reader, brings it all together

intercepts

ven centra los numeros de esta tabla horizontalmente

Aca deberia de decir es Lun, Mar, Mier

Aca deberia de decir es Lun, Mar, Mier

Aca hagamos que en los hr heads solo aparesca el nombre osea en ves de Vinicius Leite diga Vinicius

Ven en estas tablas de alarmas deberiamos de poner una columna al final del HR Head, pero recortando a solo el nombre en ves de Vinicius Leite poner solo Vinicius

Ojo esta tabla se debe de ordenar por los sin plan

A estas tablas que tienen saving deberian de tener una fila final de total

saca este signo de USD que ya se entiende de por si que son USD de todas las tablas

Ven deberiamos tener una columna antes que sea la columna con plan osea los 1.4M y al final una con el total de los 2.1M

Aca falta poner la columna de el total de savings que debe cada hr head del punto 2 como primer columna.

Y despues de esa columna deberia ir Atrasadas, Canceladas, Licencias, Sin HR, Sin Link.

❯ te falto centrar horizontalmente todas las columnas de todas las tablas excepto por la primer columna de todas las tablas

Ven en todas las tablas las columnas despues de la primer columna deberian de tener todas su contenido centrado horizontalmente.

Y en si todas las tablas deberian de tener sus filas centradas horizontalmente

Ven el orden de las alertas deberia ser primero las rojas y despues las amarillas

esta tabla solo mostremosla en el HTML no en el mensaje principal

ven estos numeros deberian de estar en millones ej: 0,70. Asi entra todo mucho mas

Esta tabla mejor armemosla con:

Filas: Contracting Country (columna J del file) Columnas: Cantidad de casos por día.

En el HTML si podriamos poner un apendice con el detalle de los casos en formato tabla

Esta tabla mejor armemosla con:

Filas: Contracting Country (columna J del file) Columnas: Cantidad de casos por día de notificación

En el HTML si podriamos poner un apendice con el detalle de los casos en formato tabla

Actua como un asesor senior de mckinsey company. Cada sección del html y del mensaje de slack deberia de tener un titulo activo de lo que se quiere transmitir

Aca falta poner una columna del saving que esta en riesgo columna W del file de transformación

Aca falta poner una columna del saving que esta en riesgo columna W del file de transformación

Aca falta poner una columna del saving que esta en riesgo columna W del file de transformación

saca esta columna ya se entiende con el titulo de la seccion

Todo lo que sean columnas que traingan fechas deberian de seguir el formato dd/mm/aa

hagamos que esta columna se vea asi dd/mm/aa

A estas tablas les falta la ultima fila de total

este % no dice nada

Aca deberia de decir de los 2.1M nos faltan 700k y sacar la bola de la derecha

Esta tabla deberia de tener solo la columna c) Savings w/o Plan del reporte de excel e ir validando como evoluciona dia a dia.

Esto sacalo, pero pon estos circulitos en la tabla de arriba para marcar cuales estan mejor y peor

saca esto para que las columnas de la derecha tengan mas espacio

haiku gets it right

haiku gets this right - 9 Nov 2026, incognito mode

While the treaty was harsh, it could be modified in the future to account for any changes in Germany's economic, political, etc. outlook. It was therefore at the Wests discretion to modify the terms to allow for the best outcome. It was ultimately a lack of Western intervention and sustained European crisis which led to a break down of the peace treaties

US influence was dramatically weakened by the refusal to ratify the Versailles Treaty

Wilson sought to prevent a full discrimination of Germany through US influence in the League and Allied economic dependence on the US

French leaders conceded in the hope that postwar plebiscites would ensure French control of the Saarland and Rhineland (along with German failures of reparation payments)

Treaty of Versailles came out of the Allied determination to dramatically weaken Germany and to prevent her return to great power status

Wilson merely sought German alignment on the new order in order to welcome Germany back into the international order

Anglo-French leaders sought to exclude Germany from the postwar order to prevent a return of German influence in Europe

Powerful states which could contain Germany (ethnic coherence was ignored in favor of balance of power concerns)

France sought an intact Austria-Hungary and Greater Poland which could serve as a check to German power (balance of power concerns at the forefront)

France feared that Wilsonianism would actually leave a fractured Central Europe which Germany could easily take advantage of, which would leave France much less secure

The Allies would now use any tools to weaken German influence

German influence at its greatest territorial extent, could not be allowed to persist in any way, out of fear of its imperialistic ambitions (as shown in the Brest-Litovsk Treaty)

German Military Imperialism spurred the Allies to expand and toughen their war aims against the Central Powers

US toughness would undercut this strategy, a position which would only be realized too late amongst the German leaders

Baden sought to use the US to undercut the Anglo-French and thus secure more lenient terms

Germany saw the 14 Points as reasonable and could serve as an ideal position for negotiations (however, their hopes of a negotiations along the 14 points would be shattered in 1919)

The failure of decisive victory, and thus discrediting of the German military leaders, meant that the position of negotiated peace finally won out in Germany

Bolshevik cooperation was seen as critical to prevent large Allied demands postwar

Civilian leaders however, saw the victory in the East as an essential negotiating point which would ensure favorable peace terms (with the US joining the war, decisive victory was seen as impossible)

Military leaders saw the victory in the east as the chance to finally secure their decisive victory and achieve maximum gain

Germanies victory in the East dramatically escalated, German territorial ambitions

Wilson understood the need to compromise and allow for a convergence of Allied war aims which could secure a definite (though not permanent) peace.

Fourteen Points clashed with the War Aims of the British and French and led to considerable conflict and debate between the Three Great Powers after WW1

Germany could retain its influence as long as it submitted to US ideals (and US dominance) in the postwar era

US would advance war aims that, outside traditional European power politics, could be delineated along principled lines

US would operate as an independent power, not subject to European power wrangling

Wilson saw the spread of US values as key to creating a new world order which would prevent costly wars and protect US interests

US threatened by European war, would need to take a more active role in the world in order to secure US interests and prevent war

Wilson saw a renewed Monroe Doctrine which would strengthen the US block which could rival the European blocks

Wilson wanted to see the Allies gain ascendency to create a new world order, which the former Central Powers could be included into (should they meet the necessary conditions)

1918 saw a toughening stance as both sides, increasingly desperate to achieve victory, went all out in their determination for victory and thus saw a subsequent toughening of their war aims

Military victory led to increased war aims

While negotiated peace was possible neither side wanted to strengthen its enemies and therefore leave itself weakened in return

Negotiated peace became more appealing as leaders struggled to achieve a decisive victory

Annexation was rejected as policy as self-determination and popular rule overcame foreign rule of self determining polities (this however did not apply to colonial subjects)

France rejected conquest but still wished for guarantees of security (land claims, reparations, demilitarization, etc?)

War aims now began to be influenced by ideological concerns as public opinion held greater sway on the articulation of the war aims

Traditional power politics at work prior to 1917

Wilsonianism would become greatly influential as the Allies became more dependent on US support

British policy of nationalist promises would turn on French and British Imperial Ambitions in the Postwar era

Imperial Conflict and Rivalry between Britain and France would have consequences in the post-war future

British war aims were largely colonially motivated (as well as maintaining its honor and moral position through the restoration of Belgium)

Britain saw German Central European control and that only be restricting German influence to Europe would prevent it from becoming a true superpower

Britain was committed to the war in 1916 and sought to ensure the completion of its war aims through the support of the United States (though this would leave the United States in a powerful position vis a vis Britain)

French and Britain would work together to manage raw material imports in order to restrict German influence

French customs union would serve to retain French western continental dominance

France saw Germany's lands as necessary to ensure French economic equality to Germany and Britain

Strategic security: military and political advantage over Germany through buffer zones, strategic alliances, and the substantial weakening of the German polity Economic security: economic advantage over Germany through the seizure of wealth lands (to make up for French losses), reparations, and the weakening of the German economy

French obsession with national security especially due to the German threat

France wanted a clean slate to deal with the German Problem as it pleased (after the disaster of 1870), in order to ensure French national security and territorial ambitions

Britain sought to limit German influence through the seizing of German colonial possessions and by limiting its military force

French would need guarantees, most notably of land, to ensure their security in any peace deal

Creation of a greater France at the expense of Germany

War was fought not merely to defend French territory (or restore French territorial integrity) but also to destroy the unity of the German Reich (e.g. land seizures)

Allies were far more determined to curb the influence of Germany than simply defend their own influence

While the military leaders were influential, it wasn't until the smashing victories in the East that their vision of maximum expansionism was legitimated

Military leaders pushed to use all options at their disposal to win a victory in the West (and secure maximum gain) while the diplomats pushed for reason

With separate peace out of the question, Germany decided to expand its aims to protect its domestic legitimacy

Separate peace failed as the Allies sought to end the threat of German militarism permanently

Land claims would therefore need to be kept reasonable in order to entice an Allied power to sue for a separate peace

Mitteleeuropa would depend on victory over the Allies

Customs Union was seen as a way of preserving German control without resorting to massive annexations which would only further antagonize her enemies in a time where total victory had slipped away from her

Expansion largely escaped the controls of the diplomats as the generals took control

Ambitions were both restricted and expanded by the war

Germany was motivated primarily by continental Imperial Aims according to Fischer as defined by the September programme

A supprimer

A supprimer

Sauter une ligne pour bien distinguer les auteurs des victimes

A supprimer

à supprimer

à supprimer

A enlever.

de mes honoraires

et selon l'affaire avec un honoraire de résultat.

Pour poser vos questions sur le suivi d'un dossier, me transmettre les documents nécessaires. Le contact par mail est à privilégier en dehors des situations d'urgence.

enlève ce qu'il y a dans les parenthèses. Pour une urgence, appelez-moi directement.

Prenez un rendez-vous via le lien ci-dessous, en personne ou par téléphone. Je ne manquerai pas de valider la date choisie en fonction de mes disponibilités ou de vous en proposer une nouvelle au plus tôt.

A enlever

J'enlèverai cette partie là. Juste le serment, j'aime bien

Je me pose toujours la question de cette photo avec le halo lumineux autour... C'est bof. Parce que sur mon parcours la photo fait bien. Elle est avec son cadre et derrière c'est en crème. Soit on renonce à cette photo (est ce qu'une autre photo ferait mieux) soit on mets autre chose ou rien.

A enlever

Sometimes they are a c name

Do we need this here it just takes space on the screen and it's like an actual thing to read if we're gonna add like sections will one we should note them down so the YouTube we can do like the screens or the chapters or whatever but I don't know if it should be on the screen that long or it should be there at all

We should talk about and say authentication not it

I don't think people aren't gonna understand what it means on its own name so we might say authenticates it's emails coming from the same domain I don't know more clear obviously

Whenever this like part is finished we need to say like and let me explain to you why very simply or whatever simplified version whatever

And it reports nothing back to you letting you know that somebody is pretending to use to be you by using your domain or subdomains

They will call it a day and he will forget about it forever I don't know how to say that without sounding too sarcastic but it's true

Demar has been around for over a decade or whatever it is.

It's not your fault you weren't aware or you were only recently aware and if your new business you might've heard the word and still not sure what to do with it

eLife Assessment

This is a useful study that seeks to elucidate the molecular mechanisms underlying spinal circuit assembly. The authors demonstrate that Onecut transcription factors in spinal motor neurons control the development of pre-motor interneurons in a non-cell autonomous manner. However, certain conclusions are incompletely supported by the data at hand. The work will be of broad interest to cell and developmental biologists.

Reviewer #1 (Public review):

Summary:

This is a useful study by Angla et al that investigates the basis of observations made from previous studies where loss of Onecut (OC) transcription factors lead to changes in spinal interneuron populations that do not themselves normally express OC. The authors hypothesize that OC expression in spinal motor neurons has non-cell-autonomous effects on pre-motor interneuron (V1, V2a/b/c) population size and distribution. By knocking out OC in the motor neuron lineage (i.e., downstream of Olig2, a motor neuron progenitor marker gene), they indeed show that motor neuron-specific loss of OC expression decreases V2c interneuron number and alters the spatial distribution of V1, V2a, V2b, V2c populations. Using bulk RNA-sequencing of WT and OC conditional knockout (cKO) motor neurons, the authors identify that neurotrophic factor Ntf3 as another factor that is responsible for the non-cell-autonomous effects on the interneuron population; upon knock out of Ntf3 downstream of Olig2 they show that this leads to increased interneuron numbers and alters their spatial distribution, ultimately leading to dysregulation of spinal motor circuits and motor activity.

Strengths:

The authors use sophisticated genetic tools to precisely remove OC and Ntf3 expression in a lineage-specific manner and comprehensively assess the downstream effects across brachial, thoracic, lumbar levels of the spinal cord, as well as at two developmental timepoints, E12.5 and E14.5.

Weaknesses:

The evidence supporting their claims is incomplete. In figure 2 and 4, the control distributions (despite reporting the same populations in the same regions) look different, suggesting large sample-to-sample variance in distribution. The authors explain that this is due to several technical limitations in their study.

Furthermore, the way the Ntf3 results are framed can be misleading; it reads as though one of the main claims of this study is that OC acts through Ntf3.

Reviewer #2 (Public review):

The study by Angla et al. proposes a model in which NT-3 produced by motor neurons regulates interneuron numbers and distribution in a non-cell autonomous manner. The authors demonstrate that ablation of motor neurons (MNs) and global and conditional deletion of OC transcription factors lead to changes in interneuron distribution. They identify that NT3 is upregulated after MN-specific OC deletion in RNA-seq experiments and show that olig2-cre mediated NT3 deletion leads to increased ventral interneuron numbers, altered distribution and defects in locomotor behavior. The authors conclude that MN-derived NT3, under OC control, regulates interneuron development. While this is an intriguing hypothesis, the study does not consolidate the results from the different experimental models in a cohesive model to provide any mechanistic insights into how NT3 may accomplish this function.

(1) The authors utilize different experimental models (OC transcription factor global and conditional knockouts, DTA ablation and NT3 deletion) to show that MNs may influence interneuron numbers and positioning, but there doesn't seem to be a direct link into how each manipulation relates to the others in an informative way. For example, the relationship between OC and NT3 deletion data is not entirely clear, even though NT3 is downstream of OC transcription factors. Both deletions lead to changes in interneuron distribution but there doesn't seem to be any correlation between the two that relates to relative changes in NT3 levels. Although it is understandable that transcription factor manipulations may have additional effects, it is hard to derive any mechanistic insights into how exactly MNs influence interneuron differentiation or migration from the data presented here. Similarly, while the rationale for following up on NT3 seems to be the chick electroporation experiments, only a very minor decrease in Chx10 interneurons is shown in those experiments. Shouldn't NT3 overexpression lead to substantial decreases in IN numbers according to the authors' model?

(2) The study primarily quantifies interneuron numbers and distribution at different levels of the spinal cord and under different genetic manipulations. In different figures, the values and distributions shown for controls vary quite a lot. For example, in figure 2B vs figure 4B, the number of FoxP2+ V1 neurons at brachial levels is ~350 vs 125. Similarly, the control distributions in 2I and 4I are quite different. While this can be partially explained by technical issues, it remains challenging to determine whether the conclusions regarding the impact of each genetic manipulation on interneuron numbers and distribution are valid.

(3) It is not clear that the behavioral phenotypes seen in the olig2-cre mediated deletion of NT3 can be attributed to specific changes in interneuron development, connectivity or function. While the behavioral phenotype is in theory consistent with defects in motor circuits, there is a big gap between the embryonic changes shown here and behavior, with no in between circuit level changes in locomotor circuits shown. Thus, the behavioral experiments are not very informative as to how exactly components of motor circuits might be altered with the NT3 deletion.

Reviewer #3 (Public review):

This manuscript aims to investigate cell extrinsic mechanisms that regulate differentiation and distribution of interneuron types in the spinal cord. The authors demonstrate that the loss of motor neurons leads to changes in number and distribution of different interneuron types, specifically V0v, V1, and V2b (but not V2a). The authors then hypothesize that this phenotype may be controlled by the action of Onecut (OC) transcription factors in motor neurons. Conditional knockout of OC1 + OC2 in motor neurons using Olig2-Cre, however, does not lead to significant changes in numbers of V1, V2a, V2b interneurons, although there is a change in their spatial distribution. While the authors do not check V0v neurons in OC mutants, they do check V2c, which show a reduction in number and change in distribution. Why the same neurons are not checked across experiments is unclear. The authors then analyze existing RNA-seq data to identify factors that could be mediating effects of the OC factors in motor neurons. They identify Ntf3 as a candidate and confirm that it is upregulated in OC mutants. Conditional loss of function of Ntf3 (Olig2-Cre) leads to increases in V1, V2a, and V2b (but not V2c) interneurons and changes in distribution of all four interneuron types. However, the Ntf3 phenotypes do not necessarily suggest that it works in the same pathway as OC to control interneuron development. Finally, the authors demonstrate that these Ntf3 conditional mutants have major defects in motor function.

The main conclusions of the manuscript are that i) motor neurons non-cell autonomously control interneuron development, ii) that OC activity in Olig2 lineage cells non-cell autonomously controls aspects of interneuron development, and iii) that Ntf3 in motor neurons non-cell autonomously controls aspects of interneuron development. The mechanistic link between OC and Ntf3 activity is not clear. And the interneuron phenotype seen when motor neurons are depleted it not phenocopied by either loss of OC or Ntf3 from motor neurons. While the authors addressed some of my initial comments, the impact of this work on the field remains difficult to assess for the following reasons:

(1) The manuscript relies heavily on quantifying numbers and spatial distribution of interneuron populations. However, these do not seem to be consistent in control animals across experiments, making it difficult to interpret any changes observed in genetic manipulations. Specifically, in Fig. 2 and 4, the same markers are being used to quantify V1, V2a, V2b, and V2c interneurons in controls vs. OC (Fig. 2) or Ntf3 (Fig. 4) conditional knockouts, but the numbers of neurons and their distribution in control animals are variable between these two figures. For example, there seem to be a mean of >300 V1 neurons in E12.5 brachial sections of Fig. 2 controls, but this number is <150 in Fig. 4 controls. The cell distribution scoring is similarly variable between these controls without any explanation. The same is true for E14.5 controls used in Fig. S2 vs. Fig. S6.<br /> The authors explain in response to the 1st review that technical variability including different antibodies, different researchers performing experiments, and different mouse backgrounds may contribute to this variation, but do not include this explanation clearly in the manuscript where it would be useful for readers. It would have also helped, for example, if authors had labeled datapoints on bar plots based on the animal that they came from to understand inter-animal variability within experiments. Overall, the biological significance of the results remain difficult to interpret due to high technical variability.

(2) In the initial review, it was noted that there is no data to support the claim that OC1/2 mutants have altered interneuron development because of higher Ntf3 expression. In response to this, the authors note that the manuscript does not claim that OC exerts its non-cell autonomous effects through Ntf3 and add a new paragraph in the discussion which now addresses the fact that it is difficult to compare the phenotypes observed the different genetic manipulations presented in the work. While this paragraph addresses the incomplete mechanistic link between OC and Ntf3, it also decreases the mechanistic impact of the work.

(3) In Figure 3E-3G, the authors show that some V1, V2a, V2b interneurons express the Ntf3 receptor TrkC. It is not stated (apologies if I missed it) what age this expression is, and from the images, it looks like in addition to medially located newborn neurons being negative, laterally located Gata3 interneurons are also broadly negative for TrkC, and many Foxd3 and Chx10 interneurons are also negative. Separate images of each channel and/or scoring would help clarify this. If many relevant interneurons are indeed TrkC negative, how this affects the conclusions should be addressed in the manuscript.

(4) Behavioral phenotypes are not necessarily caused by Ntf3's role in premotor interneuron development.

Author response:

The following is the authors’ response to the original reviews.

Public Reviews:

Reviewer #1 (Public review):

Summary:

In this study, Angla et al investigate the basis of observations made from previous studies where loss of Onecut (OC) transcription factors leads to changes in spinal interneuron populations that do not themselves normally express OC. The authors hypothesize that OC expression in spinal motor neurons has non-cell-autonomous effects on pre-motor interneuron (V1, V2a/b/c) population size and distribution. By knocking out OC in the motor neuron lineage (i.e., downstream of Olig2, a motor neuron progenitor marker gene), they indeed show that motor neuron-specific loss of OC expression decreases V2c interneuron number and alters the spatial distribution of V1, V2a, V2b, and V2c populations. Using bulk RNA-sequencing of WT and OC conditional knockout (cKO) motor neurons, the authors identify that the neurotrophic factor Ntf3 is downregulated by OC expression. They subsequently hypothesize that the non-cell-autonomous effects observed by loss of OC expression in motor neurons can be explained by de-repression of Ntf3. To test this, the authors conditionally knock out Ntf3 downstream of Olig2 and show that this leads to increased interneuron numbers and alters their spatial distribution, ultimately leading to dysregulation of spinal motor circuits and motor activity.

Strengths:

The authors use sophisticated genetic tools to precisely remove OC and Ntf3 expression in a lineage-specific manner and comprehensively assess the downstream effects across brachial, thoracic, lumbar levels of the spinal cord, as well as at two developmental timepoints, E12.5 and E14.5.

Weaknesses:

There are two main concerns that are not fully addressed:

(1) Based on the effects observed with OC vs. Ntf3 cKO, it is unclear whether OC is indeed exerting its non-cell-autonomous effects via Ntf3. Knocking out both Ntf3 and OC and comparing the effects to those seen with just OC cKO alone could provide more insight on this point.

In this study, we did not intend to demonstrate that Onecut transcription factors exert their non-cell autonomous action on spinal interneuron development by regulating Ntf3 expression, and we do not state in the manuscript that this is the case. We only show that Onecut factors and Ntf3, the expression of which they regulate, contribute to the non-cell autonomous regulation of spinal interneuron development by the motor neurons. We are convinced that Onecut factors could regulate multiple independent factors and pathways involved in extrinsic regulation of interneuron development. This possibly also includes, as demonstrated in cell culture for multiple homeoproteins including human Onecut factors, the intercellular transfer of the Onecut homeoproteins during spinal cord development, a process that we are currently investigating. Knocking out both OC and Ntf3 in the motor neurons, beyond being technically extremely challenging (1/64 probability to obtain double-mutant embryos), would not enable to address this question, as it will simply results in the addition of two different defects.

Also, a quantitative summary of the effects of Ntf3 overexpression in motor neurons in the chick is lacking.

A quantitative summary of the effects of Ntf3 overexpression in the chicken embryonic spinal cord is provided in Figure S2.

(2) How the authors assess changes in the spatial distribution of interneurons is unclear. In Figures 2 and 4, the control distributions (despite reporting the same populations in the same regions) look different, suggesting large sample-to-sample variance in distribution. Although the authors report that several sections in each level were taken from at least three animals for each condition, it's unclear how variance within WT or cKO sections was accounted for in the final statistical evaluation. It seems at a glance that a comparison between control samples in Figure 2 and Figure 4 could report statistically significant differences, which would be problematic. A more rigorous report of sample-to-sample variance and a more in-depth explanation of the statistical methods are needed.

The experimental procedure to analyze the spatial distribution of spinal interneurons at different stages of development is described in details in the “Statistical analyses” paragraph of the Materials and Methods section of the manuscript, and has been repeatedly used by ourselves [1,2] and by others (see for example [3-5]) to conduct similar analyses.

We also noticed that the distribution of the different analyzed interneuron populations in the control embryos showed some differences between the cOc1Oc2^-/- and in the cNtf3^-/- lines. Several parameters can account for this observation. First, this study has been conducted over a period of 15 years, different investigators each contributing to different steps of the analysis. Second, the genetic background of these two lines is not identical, impacting both the duration of the gestation (hence, the embryonic stage of the performed analyses, even if the embryos were collected on the same gestation day) and possibly the distribution of some interneuron populations. Third, because of evolutions in the availability of the primary antibodies used to label the interneuron populations of interest, the same antibodies were not used throughout the study, as stated for in the Materials and Methods section for Chx10, Gata3 and Sox1, although the same antibody was used by the same investigator to label the same interneuron population in each mouse line at each developmental stage.

A detailed description of the number of sections and embryos included in each analysis as well as the whole statistical workflow that was used for the distribution analyses, which takes into account variance within control or mutant samples, is now provided in the revised version of the manuscript.

Reviewer #2 (Public review):

The study by Angla et al. proposes a model in which NT-3 produced by motor neurons regulates interneuron numbers and distribution in a non-cell autonomous manner. The authors demonstrate that ablation of motor neurons (MNs) and global and conditional deletion of OC transcription factors lead to changes in interneuron distribution. They identify that NT3 is upregulated after MN-specific OC deletion in RNA-seq experiments and show that olig2-cre mediated NT3 deletion leads to increased ventral interneuron numbers, altered distribution, and defects in locomotor behavior. The authors conclude that MN-derived NT3, under OC control, regulates interneuron development. While this is an intriguing hypothesis, additional experiments are needed to support it and strengthen the link between the different experiments described here.

(1) The study primarily quantifies interneuron numbers and distribution at different levels of the spinal cord and under different genetic manipulations. Experimental details are lacking, defining how many sections were analyzed (several are noted in the methods) and how the rostrocaudal levels of the spinal cord were precisely aligned.

A detailed description of the number of sections and embryos included in each analysis as well as the whole statistical workflow that was used for the distribution analyses is now provided in the revised version of the manuscript. The rostrocaudal levels of the spinal cord were precisely aligned using the distribution of Foxp1 in the Lateral Motor Columns (LMCs) at brachial or lumbar levels of the spinal cord, which is also indicated in the revised version.

In different figures, the values and distributions shown for controls vary quite a lot. For example, in Figure 2B vs Figure 4B, the number of FoxP2+ V1 neurons at brachial levels is ~350 vs 125. Similarly, the control distributions in 2I and 4I are quite different. This makes it challenging to determine whether the conclusions regarding the impact of each genetic manipulation on interneuron numbers and distribution are valid.

Several parameters can account for these observations. First, this study has been conducted over a period of 15 years, different investigators each contributing to different steps of the analysis. Second, the genetic background of these two lines is not identical, impacting both the duration of the gestation (hence, the embryonic stage of the performed analyses, even if the embryos were collected on the same gestation day) and possibly the distribution of some interneuron populations. Third, because of evolutions in the availability of the primary antibodies used to label the interneuron populations of interest, the same antibodies were not used throughout the study, as stated for in the Materials and Methods section for Chx10, Gata3 and Sox1, although the same antibody was used by the same investigator to label the same interneuron population in each mouse line at each developmental stage.

(2) The relationship between OC and NT3 deletion data is not entirely clear. Both deletions presumably lead to changes in interneuron distribution, but is there any reverse relationship between the two that relates to relative changes in NT3 levels? The authors do not directly compare NT3 and OC KO IN distributions. Similarly, one might expect a decrease in interneuron numbers in OC mutants, which is only reported for V2c neurons. However, the image presented in Figure 2G shows an equal number of V2c INs in control and mutant.

In this study, we did not intend to demonstrate that Onecut transcription factors exert their non-cell autonomous action on spinal interneuron development by regulating Ntf3 expression, and we do not state in the manuscript that this is the case. We only show that Onecut factors and Ntf3, the expression of which they regulate, contribute to the non-cell autonomous regulation of spinal interneuron development by the motor neurons. We are convinced that Onecut factors could regulate multiple independent factors and pathways involved in extrinsic regulation of interneuron development. For example, as mentioned above, we observed intercellular transfer of the Onecut homeoproteins during spinal cord development, suggesting different strategies for non-cell autonomous regulation of interneuron development. The two mouse lines studied here consist on the one side in a combination of OC inactivation and Ntf3 increased expression, and on the other side in Ntf3 inactivation. Therefore, a reverse relationship between the changes in interneuron distribution is not expected. Furthermore, gain-of-function and loss-of-function experiments in mouse models frequently generate phenotypes that are not inverse to each other.

(3) It is not clear that the behavioral phenotypes seen in the olig2-cre mediated deletion of NT3 can be attributed to changes in interneuron development. How about a role of NT3 in oligodendrocytes? There is a big gap between the embryonic changes shown here and behavior, with no in-between circuit-level changes in locomotor circuits shown.

We agree, the motor behavior changes that we recorded in Ntf3 conditional mutant mice are, as stated, “consistent with the hypothesis that Ntf3 produced by MNs is required to generate locomotor circuits with properly coordinated activity” but do not demonstrate a direct causal relationship. However, investigating the intrinsic activity of the spinal locomotor circuits, independently from, for example, oligodendrocyte contribution may prove to be extremely challenging and was beyond the scope of this study. In addition, to our best knowledge, Ntf3 has not been shown to be expressed in healthy oligodendrocytes in vivo, and TrkC has not been reported to be displayed by these cells in the same conditions.

A more restricted manipulation would be deleting TrkC from specific interneuron populations. Related to this, although TrkC is shown to be broadly expressed in ventral interneurons, it is not shown specifically to colocalize with any of the interneuron markers. The authors should validate that the receptor is expressed in the subsets that they are investigating.

We agree, investigating the consequences of inactivating the TrkC receptor in specific interneuron populations would be extremely informative. However, this experiment is also very challenging to perform, as most of the driver lines available to target spinal interneuron populations additionally target multiple neuronal populations outside of the spinal cord that are also involved in the control of movements and could therefore induce confounding effects on motor behavior analyses.

We thank the reviewer for suggesting to investigate in more details the interneuron populations that display TrkC receptors, which is now included in the revised version of the manuscript (Figure 3E-G).

(4) The rationale for following up on NT3 seems to be the chick electroporation experiments; however, no changes in distribution are shown in those experiments, and only a very minor decrease in Chx10 interneurons. Shouldn't NT3 overexpression lead to substantial decreases in IN numbers according to the authors' model? The "data not shown", which presumably refers to distribution, would be important to show here, to further support this rationale.

Chicken spinal cord electroporation only enables to study spinal cord development for a few days, given the high mortality rate observed after longer incubation. At the stage we collected the electroporated embryos for analyses, interneuron migration has barely been initiated, and distribution cannot be studied yet. Consistently, we are not aware of any report of interneuron distribution analysis in electroporated chicken embryonic spinal cord, as compared to mouse embryos [1-5].

(5) The idea that NT3 downregulation causes an increase in IN numbers is not intuitive. Also, considering the DTA experiments in Figure 1, showing that MN ablation leads to a decrease in several IN subtypes and no changes in V2a neurons. It would be helpful for the reader if the authors could synthesize their results in the discussion and reconcile their experimental findings.

We agree, this has now been included in the Discussion section of the revised manuscript.

Reviewer #3 (Public review):

This manuscript aims to investigate cell extrinsic mechanisms that regulate the differentiation and distribution of interneuron types in the spinal cord. The authors demonstrate that the loss of motor neurons leads to changes in the number and distribution of different interneuron types, specifically V0v, V1, and V2b (but not V2a). The authors then hypothesize that this phenotype may be controlled by the action of Onecut (OC) transcription factors in motor neurons. Conditional knockout of OC1 + OC2 in motor neurons using Olig2-Cre, however, does not lead to significant changes in the numbers of V1, V2a, and V2b interneurons, although there is a change in their spatial distribution. While the authors do not check V0v neurons in OC mutants, they do check V2c, which show a reduction in number and change in distribution. Why the same neurons are not checked across experiments is unclear. The authors then analyze existing RNA-seq data to identify factors that could be mediating the effects of the OC factors in motor neurons. They identify Ntf3 as a candidate and confirm that it is upregulated in OC mutants. Conditional loss of function of Ntf3 (Olig2-Cre) leads to increases in V1, V2a, and V2b (but not V2c) interneurons and changes in the distribution of all four interneuron types. Finally, the authors demonstrate that these Ntf3 conditional mutants have major defects in motor function.

The conclusions of this manuscript are not well supported by the data for the reasons listed below, making it difficult to assess the impact of this work on the field.

(1) The manuscript relies heavily on quantifying numbers and the spatial distribution of interneuron populations. However, these do not seem to be consistent in control animals across experiments, making it difficult to interpret any changes observed in genetic manipulations. Specifically, in Figures 2 and 4, the same markers are being used to quantify V1, V2a, V2b, and V2c interneurons in controls vs. OC (Figure 2) or Ntf3 (Figure 4) conditional knockouts, but the numbers of neurons and their distribution in control animals are variable between these two figures. For example, there seems to be a mean of >300 V1 neurons in E12.5 brachial sections of Fig. 2 controls, but this number is <150 in Fig. 4 controls. The cell distribution scoring is similarly variable between these controls without any explanation. The same is true for E14.5 controls used in Figure S1 vs. Figure S3.

We also noticed that quantifications and distributions of the different analyzed interneuron populations in the control embryos showed some differences between the cOc1Oc2^-/- and in the cNtf3^-/- lines. Several parameters can account for this observation. First, this study has been conducted over a period of 15 years, different investigators each contributing to different steps of the analysis. Therefore, the interneuron distribution analyses in these two mouse lines were not conducted by the same researchers. Second, the genetic background of these two lines is not identical, impacting both the duration of the gestation (hence, the embryonic stage of the performed analyses, even if the embryos were collected on the same gestation day) and possibly the distribution of some interneuron populations. Third, because of evolutions in the availability of the primary antibodies used to label the interneuron populations of interest, the same antibodies were not used throughout the study, as stated in the Materials and Methods section for Chx10, Gata3 and Sox1, although the same antibody was used by the same investigator to label the same interneuron population in each mouse line at each developmental stage.

(2) Neurotrophic factors generally promote neuronal survival. However, in this study, the loss of Ntf3 leads to increased numbers of interneurons. This finding is in disagreement with previous observations in slice cultures of spinal cords, as stated in the discussion. This discrepancy makes it even more important that the cell counts reported in the figures discussed above are robust.

Considering that neurotrophic factors only support neuronal survival would strongly neglect their important function in neuronal differentiation, which has been broadly demonstrated. Severe immunotoxic ablation of motor neurons or anti-serum blockade of Ntf3 activity severely depleted inhibitory, but not excitatory, interneurons in a highly apoptotic-prone organotypic culture model of embryonic rat spinal cord slices, which was rescued by Ntf3 in the first model [6]. Opposite results were obtained in vivo by other researchers using mouse models lacking almost all MNs due to the elimination of skeletal muscles, where the number of spinal INs remained unaffected [7,8]. Combined to our results, these in vivo observations suggest that Ntf-3 is involved in interneuron differentiation rather in their survival. Consistently, Ntf3 has been shown to promote neuronal differentiation [9].

(3) The claim that phenotypes are non-cell autonomously driven by motor neurons is not well supported. In Olig2-Cre conditional knockouts of Onecut and Ntf3, there is no confirmation that the loss of these factors is specific to motor neurons. Therefore, it cannot be ruled out that other cell populations may be mediating the phenotypes.

Combined conditional inactivation of Oc1 and Oc2 has been reported in [10]. Conditional inactivation of Ntf3 only impacts motor neurons as it is the only cell population in the ventral spinal cord wherein this factor is produced (this study and [11-13]). Furthermore, Olig2-Cre has been shown to be active in motor neurons and in V3 interneurons (see for example [14], which, for this reason, have not been studied in the frame of this project as stated in the manuscript.

(4) The claim that interneuron development is regulated by OC control of Ntf3 expression in motor neurons is not well supported. The authors show that loss of OC1/2 leads to an increase in Ntf3 expression in motor neurons. If this pathway were controlling interneurons, loss of OC function and overexpression of Ntf3 would have the same phenotype, which is not the case. Additionally, it would also be expected that loss of OC function and loss of Ntf3 function would have inverse phenotypes, which is also not the case. The phenotypes from OC loss of function and Ntf3 loss of function seem distinct from one another. The authors state that too little and too much Ntf3 are both bad for interneuron development, but there is no data to support their claim that OC1/2 mutants have altered interneuron development because of higher Ntf3 expression.

In this study, we did not intend to demonstrate that Onecut transcription factors exert their non-cell autonomous action on spinal interneuron development by regulating Ntf3 expression, and we do not state in the manuscript that this is the case. We only show that Onecut factors and Ntf3, the expression of which they regulate, contribute to the non-cell autonomous regulation of spinal interneuron development by the motor neurons. We are convinced that Onecut factors could regulate multiple independent factors and pathways involved in extrinsic regulation of interneuron development. This possibly also includes, as demonstrated in cell culture for multiple homeoproteins including human Onecut factors, the intercellular transfer of the Onecut homeoproteins during spinal cord development, a process that we are currently investigating.

(5) It is not clear that interneurons being studied express the Ntf3 receptor TrkC, which makes it difficult to assess whether changes in Ntf3 signaling are directly responsible for the phenotype.

We thank the reviewer for suggesting to investigate in more details the interneuron populations that display TrkC receptors, which is now included in the revised version of the manuscript (Figure 3E-G).

(6) While the behavioral phenotypes are consistent with Ntf3 playing a role in motor circuits, there is no evidence to suggest that Ntf3's influence on premotor interneurons being studied is driving or contributing to this phenotype, as discussed by the authors.

We agree, the motor behavior changes that we recorded in Ntf3 conditional mutant mice are, as stated, “consistent with the hypothesis that Ntf3 produced by MNs is required to generate locomotor circuits with properly coordinated activity” but do not demonstrate a direct causal relationship. However, investigating the intrinsic activity of the spinal locomotor circuits was beyond the scope of this study.

Recommendations for the authors:

Reviewer #1 (Recommendations for the authors):

(1) Figure 3: A more comprehensive overview of the RNA-seq results would be helpful to see how much gene expression is shifted as a result of OC loss (e.g., volcano plot, pathway enrichment analysis).

We agree, this has now been added in Figures 3B and S3 of the revised manuscript.

(2) Figure 2, 4: How does the spatial distribution of motor neurons change?

Describing the distribution of motor neurons is highly challenging given that the expression of markers that are collectively required to label all the motor neuron subset, including Foxp1, Hb9 and most significantly Isl1, is altered when Onecut expression is modified. Furthermore, changes in motor neuron distribution could constitute consequences of alterations in pre-motor interneuron numbers or distributions. However, as far as we can observe, motor neuron distribution was not obviously modified in the studied mutants.

(3) Figure S2: Why is Ntf3 expression so limited compared to Isl1-expressing motor neurons? An explanation would be helpful.

As stated in the Material and Methods section and in the legend of Figure S4 of the revised manuscript, expression after chicken embryonic spinal cord electroporation is transient and maximal reporter gene expression, including GFP, can usually only be visualized 24h after electroporation, but not at later stages. Forty-height hours after electroporation, only residual expression can still be detected in a few targeted cells.

Reviewer #3 (Recommendations for the authors):

(1) Changes in interneuron numbers are attributed to defects in differentiation. However, the possibility that these could be caused by changes in cell death has not been ruled out.

Absence of increased cell death had been demonstrated previously for Onecut mutant embryos [15] and has been verified now in conditional Ntf3 mutant embryos (Figure S5). Both information have been included in the text.

(2) In Figures 2 and 4, it is unclear if the images of the spinal cords are from brachial, thoracic, or lumbar levels. As the three levels frequently show different phenotypes, it is difficult to match the immunostaining with the reported quantification.

All the pictures show thoracic regions of the spinal cord, as now stated in the figure legends of the revised version of the manuscript.

(3) Statistics are missing in the cell distribution plots in Figure S1.

This is not the case in our version of the pdf file.

(4) In experiments where motor neurons are eliminated with Olig2-Cre/DTA, the authors write that progenitors are not affected without showing data. These data should be included because i) Olig2 is expressed in progenitors, so it would be expected that motor neuron progenitors are lost or reduced in this mouse, and ii) showing no changes in progenitors is crucial to the claim that postmitotic motor neurons are affected interneuron populations.

We agree, these data are now included as Figure S1 of the revised version of the manuscript.

(1) Harris, A. et al. Onecut factors and Pou2f2 regulate the distribution of V2 interneurons in the mouse developing spinal cord. Front Cell Neurosci 13 (2019). https://doi.org/10.3389/fncel.2019.00184

(2) Kabayiza, K. U. et al. The Onecut Transcription Factors Regulate Differentiation and Distribution of Dorsal Interneurons during Spinal Cord Development. Front Mol Neurosci 10, 157 (2017). https://doi.org/10.3389/fnmol.2017.00157

(3) Deska-Gauthier, D. et al. Embryonic temporal-spatial delineation of excitatory spinal V3 interneuron diversity. Cell Rep 43, 113635 (2024). https://doi.org/10.1016/j.celrep.2023.113635

(4) Bikoff, J. B. et al. Spinal Inhibitory Interneuron Diversity Delineates Variant Motor Microcircuits. Cell 165, 207–219 (2016). https://doi.org/10.1016/j.cell.2016.01.027

(5) Hayashi, M. et al. Graded Arrays of Spinal and Supraspinal V2a Interneuron Subtypes Underlie Forelimb and Hindlimb Motor Control. Neuron 97, 869–884 e865 (2018). https://doi.org/10.1016/j.neuron.2018.01.023

(6) Bechade, C., Mallecourt, C., Sedel, F., Vyas, S. & Triller, A. in J Neurosci Vol. 22 8779–8784 (2002).

(7) Grieshammer, U., Lewandoski, M., Prevette, D., Oppenheim, R. W. & Martin, G. R. Muscle-specific cell ablation conditional upon Cre-mediated DNA recombination in transgenic mice leads to massive spinal and cranial motoneuron loss. Dev Biol 197, 234–247 (1998). https://doi.org/10.1006/dbio.1997.8859

(8) Kablar, B. & Rudnicki, M. A. Development in the absence of skeletal muscle results in the sequential ablation of motor neurons from the spinal cord to the brain. Dev Biol 208, 93–109 (1999). https://doi.org/10.1006/dbio.1998.9184

(9) Dutton, R., Yamada, T., Turnley, A., Bartlett, P. F. & Murphy, M. Regulation of spinal motoneuron differentiation by the combined action of Sonic hedgehog and neurotrophin 3. Clin Exp Pharmacol Physiol 26, 746–748 (1999). https://doi.org/10.1046/j.1440-1681.1999.03108.x

(10) Toch, M. et al. Onecut-dependent Nkx6.2 transcription factor expression is required for proper formation and activity of spinal locomotor circuits. Sci Rep 10, 996 (2020). https://doi.org/10.1038/s41598-020-57945-4

(11) Buck, C. R., Seburn, K. L. & Cope, T. C. Neurotrophin expression by spinal motoneurons in adult and developing rats. J Comp Neurol 416, 309–318 (2000). https://doi.org/10.1002/(sici)1096-9861(20000117)416:3<309::aid-cne3>3.0.co;2-u

(12) Henderson, C. E. et al. Neurotrophins promote motor neuron survival and are present in embryonic limb bud. Nature 363, 266–270 (1993). https://doi.org/10.1038/363266a0

(13) Usui, N. et al. Role of motoneuron-derived neurotrophin 3 in survival and axonal projection of sensory neurons during neural circuit formation. Development 139, 1125–1132 (2012). https://doi.org/10.1242/dev.069997

(14) Debrulle, S. et al. Vsx1 and Chx10 paralogs sequentially secure V2 interneuron identity during spinal cord development. Cell Mol Life Sci 77, 4117–4131 (2020). https://doi.org/10.1007/s00018-019-03408-7

(15) Roy, A. et al. Onecut transcription factors act upstream of Isl1 to regulate spinal motoneuron diversification. Development 139, 3109–3119 (2012). https://doi.org/10.1242/dev.078501

pesquisar este decreto

if i were to rewrite this paragraph i would have added some emotional descriptions, internal conflict and remove repetitive descriptions of Babamakuru's threats

i think this passage contains the main purpose or tension of the book

character struggle where Tambu struggles to her new enviroments

le lien entre apprentissage et rêve me semble particulièrement pertinent, mais dans le rêve l'investissement n'est pas financier. Médiatiser par le financier (l'argent) n'est ce pas un moyen particulièrement efficace du néo-libéralisme pour nous ligoter dans ses filets (TINA) c'est une des question que je me pose ...

Le concept de finalité ouverte me semble très intéressant en rupture lac un monde idéal transposition du retour à l'éden terrestre dont on arrive pas à se débarrasser.

Vítěz této kategorie

this is where Mumei actually gets noticed. It happens right after the map lesson, so I read it as Yonatani picking him because he’s the one who really cares about the outside world, not because he’s the strongest or smartest.

Lack of educators

Protests driving forces.

This article is published in Cellular and Molecular Life Sciences https://doi.org/10.1007/s00018-025-05730-9

Here is a template for python-lab.html, in this case minimizing \((x-2)^2 + (y-1)^2 + x y + x + y\) subject to the constraints

$$ x \leq 1, \quad y \leq 1\quad\text{and}\quad x + y \geq 1. $$

``` from scipy.optimize import minimize

def f(v): x, y = v r = (x - 2)2 + (y-1)2 + x*y +x + y return r # minimize # return -r # maximize

def g1(v): x, y = v return 1 - x # must be >= 0

def g2(v): x, y = v return 1 - y # must be >= 0

def g3(v): x, y = v return x + y - 1 # must be >= 0

cons = [ {"type": "ineq", "fun": g1}, {"type": "ineq", "fun": g2}, {"type": "ineq", "fun": g3}, ]

res = minimize(f, [1, 1], constraints=cons) print(res.x) ```

Here [1,1] is an initial point used as a guess.

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quitar guión

Wilsonian Moment and Russian Revolution combined to uplift the enemies of Empire

Western civilization and its superiority (a key part of the Empires hierarchical control) was completely destroyed after the WW1 Catastrophe