584 Matching Annotations
  1. Last 7 days
    1. Faria, N. R., Mellan, T. A., Whittaker, C., Claro, I. M., Candido, D. da S., Mishra, S., Crispim, M. A. E., Sales, F. C. S., Hawryluk, I., McCrone, J. T., Hulswit, R. J. G., Franco, L. A. M., Ramundo, M. S., Jesus, J. G. de, Andrade, P. S., Coletti, T. M., Ferreira, G. M., Silva, C. A. M., Manuli, E. R., … Sabino, E. C. (2021). Genomics and epidemiology of the P.1 SARS-CoV-2 lineage in Manaus, Brazil. Science. https://doi.org/10.1126/science.abh2644

  2. May 2021
  3. Apr 2021
    1. Dr Ellie Murray on Twitter: “There are 3 types of disaster responses: •panicking or freezing; •taking action; and •ignoring the disaster. That last one is the most common response to sudden disasters, like when, for example, a ferry sinks. I didn’t expect it would also be most common in a pandemic.” / Twitter. (n.d.). Retrieved April 25, 2021, from https://twitter.com/EpiEllie/status/1384223819670245378

    1. Mehdi Hasan. (2021, April 12). ‘Given you acknowledged...in March 2020 that Asian countries were masking up at the time, saying we shouldn’t mask up as well was a mistake, wasn’t it... At the time, not just in hindsight?’ My question to Dr Fauci. Listen to his very passionate response: Https://t.co/BAf4qp0m6G [Tweet]. @mehdirhasan. https://twitter.com/mehdirhasan/status/1381405233360814085

    1. Indian entrepreneurs found ways to set up their own modern factories to rival British products.
      • Some Indian businesspeople responded to the industrialization of the British with their own industrialization to outcompete them
    2. Hindu-Muslim unity,
      • Both Hindus and Muslims despised the British enough to work together
    3. he Rebellion of 1857—-was the “greased cartridge” controversy.
      • In 1857, a rumor was spread that the British were using cow and pig fat in the cartitriges Indian soldiers had to bite off which caused a rebellion of soliers who believed this meant the British were trying to convert them to Christianity.
      • This event demonstrates that there was massive distrust in the British as well
      • People would not start a huge rebellion based on a small rumor if they were not already angry with the status quo and were waiting for the last straw.
    4. This was a new colonial order, but it was not stable.

      Along with peasants, other sectors of the Indian population were not happy with British rule:

      • Indian people who once had much power and property
      • Indian business people who lost a lot of power in the newly British economy
    5. the uprisings were local in scale and vision.
      • The many revolts that made up the 1857 Rebellion were usually for specific villages or small areas for the inhabitants there
    6. the )»- portant role of the lower classes.
      • Peasants made a big part of the 1857 Rebellion because of their frustrations in not only the cultural rule of the British but the taxation rule and the loans they had to take out to pay taxes
    7. e petnte a government of his ow modeling it on the British administration.
      • The peasant Devi Singh made his own gov based on the British way of governing with a peasant army that went after the moneyloaners hated by peasants in debt
      • This may be evidence for the idea that the main sticking point for Indian peasants was the cultural and taxation policies of the British instead of the administrative part
    8. determined to destroy the religion
      • There was already massive distrust in the cultural/religious policies of the British before the cartridge controversy
      • It is notable that the main sticking point for the rebels was British religious enforcement, showing how displeased Indians were with British policies in the early 1800s to try to make Indians culturally British
  4. Mar 2021
    1. ReconfigBehSci. (2020, November 11). RT @EpiCOVIDCorps: The COVID Corps YouTube channel is live! Here’s who we are and what we’re about. New videos every Wednesday. Https://t.c… [Tweet]. @SciBeh. https://twitter.com/SciBeh/status/1326848746093752321

    1. reading process

      Hypothes.is and close reading are synonymous. But the idea of "response" as a concept is utterly changed by this social annotation software. Like using the video response below.

      https://youtu.be/VlGCxMDEknk

      Or this:

      Or this:

      https://soundcloud.com/hugo-kant/sets/the-point-of-no-return

    1. ReconfigBehSci. (2020, November 9). Great talk by Chiara Varazzani from the OECD on the two speed systems of policy and ‘normal’ research and the challenge that poses to pandemic response #scibeh2020 https://t.co/Gsr66BRGcJ [Tweet]. @SciBeh. https://twitter.com/SciBeh/status/1325725690935832576

    1. ReconfigBehSci on Twitter: ‘Session 1: “Open Science and Crisis Knowledge Management now underway with Chiara Varazzani from the OECD” How can we adapt tools, policies, and strategies for open science to provide what is needed for policy response to COVID-19? #scibeh2020’ / Twitter. (n.d.). Retrieved 5 March 2021, from https://twitter.com/SciBeh/status/1325720293965443072

  5. Feb 2021
    1. Andrew💙Croxford. (2020, December 3). NEW THREAD: possible development of anti-Syncytin responses after immunization with the SARS-CoV-2 spike protein-coding mRNA vaccines, based on a ‘homologous’ region shared between these proteins. [Tweet]. @andrew_croxford. https://twitter.com/andrew_croxford/status/1334593606196187136

  6. Jan 2021
    1. The relationship between cumulative CO2 emissions and temperature change is known as the “Transient Climate Response to Cumulative Emissions” (TCRE) and is a robust predictor of CO2-induced warming across a wide range of emissions levels and pathways.

      Ist für mich ein weiterer Topic bei der Darstellung der globalen Erwärmung.\(Insert LaTeX\)

  7. Dec 2020
    1. Corker, K. S., Arnal, J., Bonfiglio, D. B. V., Curran, P. G., Chartier, C. R., Chopik, W. J., Guadagno, R., Kimbrough, A., Schmidt, K., & Wiggins, B. J. (2020). Many Labs 5: Registered Replication of Albarracín et al. (2008), Experiment 7. PsyArXiv. https://doi.org/10.31234/osf.io/qzspr

  8. Nov 2020
    1. Author Response

      Summary:

      This work is of interest because it increases our understanding of the molecular mechanisms that distinguish subtypes of VIP interneurons in the cerebral cortex and because of the multiple ways in which the authors address the role of Prox1 in regulating synaptic function in these cells.

      The authors would like to thank the reviewers for their constructive comments. In response, we would like to clarify a number of issues, as well as outline how we plan to resolve major concerns.

      Reviewer #1:

      Stachiak and colleagues examine the physiological effects of removing the homeobox TF Prox1 from two subtypes of VIP neurons, defined on the basis of their bipolar vs. multipolar morphology.

      The results will be of interest to those in the field, since it is known from prior work that VIP interneurons are not a uniform class and that Prox1 is important for their development.

      The authors first show that selective removal of a conditional Prox1 allele using a VIP cre driver line results in a change in paired pulse ratio of presumptive excitatory synaptic responses in multipolar but not bipolar VIP interneurons. The authors then use RNA-seq to identify differentially expressed genes that might contribute and highlight a roughly two-fold reduction in the expression of a transcript encoding a trans-synaptic protein Elfn1 known to contribute to reduced glutamate release in Sst+ interneurons. They then test the potential contribution of Elfn1 to the phenotype by examining whether loss of one allele of Elfn1 globally alters facilitation. They find that facilitation is reduced both by this genetic manipulation and by a pharmacological blockade of presynaptic mGluRs known to interact with Elfn1.

      Although the results are interesting, and the authors have worked hard to make their case, the results are not definitive for several reasons:

      1) The global reduction of Elfn1 may act cell autonomously, or may have other actions in other cell types. The pharmacological manipulation is less subject to this interpretation, but these results are not as convincing as they could be because the multipolar Prox1 KO cells (Fig. 3 J) still show substantial facilitation comparable, for example to the multipolar control cells in the Elfn1 Het experiment (controls in Fig. 3E). This raises a concern about control for multiple comparisons. Instead of comparing the 6 conditions in Fig 3 with individual t-tests, it may be more appropriate to use ANOVA with posthoc tests controlled for multiple comparisons.

      The reviewer’s concerns regarding non-cell-autonomous actions of global Elfn1 KO are well founded. Significant phenotypic alterations have previously been reported, both in the physiology of SST neurons as well in the animals’ behavior (Stachniak, Sylwestrak, Scheiffele, Hall, & Ghosh, 2019; Tomioka et al., 2014). The homozygous Elfn1 KO mouse displays a hyperactive phenotype and epileptic activity after 3 months of age, suggesting generalcortical activity differences exist (Dolan & Mitchell, 2013; Tomioka et al., 2014). Nevertheless, we have not observed such changes in P17-21 Elfn1 heterozygous (Het) animals.

      Comparing across different experimental animal lines, for example the multipolar Prox1 KO cells (Fig. 3 J) to the multipolar control cells in the Elfn1 Het experiment (controls in Fig. 3E), is in our view not advisable. There is a plethora of examples in the literature on the effect of mouse strain on even the most basic cellular functions and hence it is always expected that researchers use the correct control animals for their experiments, which in the best case scenario are littermate controls. For these reasons, we would argue that statistical comparisons across mouse lines is not ideal for our study. Elfn1 Het and MSOP data are presented side by side to illustrate that Elfn1 Hets (3C,E) phenocopy the effects of Prox1 deletion (3G,H,I,J). (See also point 3) MSOP effect sizes, however, do show significant differences by ANOVA with Bonferroni post-hoc (normalized change in EPSC amplitude; multipolar prox1 control: +12.1 ± 3.8%, KO: -8.4 ± 4.3%, bipolar prox1 control: -5.2 ± 4.3%, KO: -3.4 ± 4.7%, cell type x genotype interaction, p= 0.02, two way ANOVA).

      2) The isolation of glutamatergic currents is not described. Were GABA antagonists present to block GABAergic currents? Especially with the Cs-based internal solutions used, chloride reversal potentials can be somewhat depolarized relative to the -65 mV holding potential. If IPSCs were included it would complicate the analysis.

      No, in fact GABA antagonists were not present in these experiments. The holding voltage in our evoked synaptic experiments is -70 mV, which combined with low internal [Cl-] makes it highly unlikely that the excitatory synaptic responses we study are contaminated by GABA-mediated ones, even with a Cs MeSO4-based solution. Nevertheless, we have now performed additional experiments where glutamate receptor blockers were applied in bath and we observe a complete blockade of the synaptic events at -70mV proving that they are AMPA/NMDA receptor mediated. When holding the cell at 0mV with these blockers present, outward currents were clearly visible, suggesting intact GABA-mediated events.

      3) The assumption that protein levels of Elfn1 are reduced to half in the het is untested. Synaptic proteins can be controlled at the level of translation and trafficking and WT may not have twice the level of this protein.

      We thank reviewer for pointing this out. Our rationale for using the Elfn1 heterozygous animals is rather that transcript levels are reduced by half in heterozygous animals, to match the reduction we found in the mRNA levels of VIP Prox1 KO cells (Fig 2). The principle purpose of the Elfn1 KO experiment was to determine whether the change in Elfn1 transcript levels could be sufficient to explain the synaptic deficit observed in VIP Prox1 KO cells. As the reviewer notes, translational regulation and protein trafficking could ultimately result in even larger changes than 0.5x protein levels at the synapse. This may ultimately explain the observed multipolar/bipolar disparity, which cannot be explained by transcriptional regulation alone (Fig 4).

      4) The authors are to be commended for checking whether Elfn1 is regulated by Prox1 only in the multipolar neurons, but unfortunately it is not. The authors speculate that the selective effects reflect a selective distribution of MgluR7, but without additional evidence it is hard to know how likely this explanation is.

      Additional experiments are underway to better understand this mechanism.

      Reviewer #2:

      Stachniak et al., provide an interesting manuscript on the postnatal role of the critical transcription factor, Prox1, which has been shown to be important for many developmental aspects of CGE-derived interneurons. Using a combination of genetic mouse lines, electrophysiology, FACS + RNAseq and molecular imaging, the authors provide evidence that Prox1 is genetically upstream of Elfn1. Moreover, they go on to show that loss of Prox1 in VIP+ cells preferentially impacts those that are multipolar but not the bipolar subgroup characterized by the expression of calretinin. This latter finding is very interesting, as the field is still uncovering how these distinct subgroups emerge but are at a loss of good molecular tools to fully uncover these questions. Overall, this is a great combination of data that uses several different approaches to come to the conclusions presented. I have suggestions that I think would strengthen the manuscript:

      1) Can the authors add a supplemental table showing the top 20-30 genes up and down regulated in their Prox1 KOS? This would make these, and additional, data more tenable to readers.

      We would be happy to provide supplementary tables with candidate genes at both P8 and P12.

      2) It is interesting that loss of Prox1 or Elfn1 leads to phenotypes in multipolar but are not present or mild in bipolar VIP+ cells. The authors test different hypotheses, which they are able to refute and discuss some ideas for how multipolar cells may be more affected by loss of Elfn1, even when the transcript is lost in both multipolar and bipolar after Prox1 deletion. If there is any way to expand upon these ideas experimentally, I believe it would greatly strengthen the manuscript. I understand there is no perfect experiment due to a lack of tools and reagents but if there is a way to develop one of the following ideas or something similar, it would be beneficial:

      We thank the reviewer for the note.

      a) Would it be possible to co-fill VIPCre labeled cells with biocytin and a retroviral tracer? Then, after the retroviral tracer had time to label a presynaptic cell, assess whether these were preferentially different between bipolar and multipolar cell types, the latter morphology determined by the biocytin fill? This would test whether each VIP+ subtype is differentially targeted.

      Although this is a very elegant experiment and we would be excited to do it, we do feel that single-cell rabies virus tracing is technically very challenging and will take many months to troubleshoot before being able to acquire good data. Hence, we think it is beyond the scope of this study.

      b) Another biocytin possibility would be to trace filled VIP+ cells and assess whether the dendrites of multipolar and bipolar cells differentially targeted distinct cortical lamina and whether these lamina, in the same section or parallel, were enriched for mGluR7+ afferents.

      We thank the reviewer for their suggestion and we are planning on doing these kinds of experiments.

      Reviewer #3:

      In this work Stachiak and colleagues investigate the role of Prox1 on the development of VIP cells. Prox1 is expressed by the majority of GABAergic derived from the caudal ganglionic eminence (CGE), and as mentioned by the authors, Prox1 has been shown to be necessary for the differentiation, circuit integration, and maintenance of CGE-derived GABAergic cells. Here, Stachiak and colleagues show that removal of Prox1 in VIP cells leads to suppression of synaptic release probability onto cortical multipolar VIP cells in a mechanism dependent on Elfn1. This work is of interest for the field because it increases our understanding of differential synaptic maturation of VIP cells. The results are noteworthy, however the relevance of this manuscript would potentially be increased by addressing the following suggestions:

      1) Include histology to show when exactly Prox1 is removed from multipolar and bipolar VIP-expressing cells by using the VIP-Cre mouse driver.

      We can address this by performing an in-situ hybridization against Prox1 from P3 onwards (when Cre becomes active).

      2) Clarify if the statistical analysis is done using n (number of cells) or N (number of animals). The analysis between control and mutants (both Prox1 and Elfn1) need to be done across animals and not cells.

      Statistics for physiology were done across n (number of cells) while statistics for ISH are done across number of slices. We will clarify this point in the text and update the methods.

      Regarding the statistics for the ISH, these have been done across n (number of slices) for control versus KO tissue (N = 3 and N = 2 animals, respectively). We will add more animals to this analysis to compare by animal instead, although we do not expect any change in the results.

      Regarding the physiology, we would provide a two-pronged answer. We first of all feel that averaging synaptic responses for each animal would hide a good deal of the biological variability in PPR present in different cells (response Fig 1), the characterization of which is integral to the central findings of the paper. Secondly, to perform such analysis asked by the reviewer one would need to obtain recordings from ~10 animals or so per condition for each condition, which, to our knowledge, is something that is not standard when utilizing in vitro electrophysiological recordings from single cells. For example, in these very recent studies that have performed in vitro electrophysiological recordings all the statistics are performed using “n” number of cells and not the average of all the cells recorded per animal collapsed into a single data point. (Udakis, Pedrosa, Chamberlain, Clopath, & Mellor, 2020) https://www.nature.com/articles/s41467-020-18074-8

      (Horvath, Piazza, Monteggia, & Kavalali, 2020) https://elifesciences.org/articles/52852

      (Haas et al., 2018) https://elifesciences.org/articles/31755

      Nevertheless, we have now re-run the analysis grouping the cells and averaging the values we get per animal, since we have obtained our data from many animals. The results are more or less indistinguishable from the ones presented in the original submission, except for on p value that rose to 0.07 from 0.03 due to the lack of the required number of animals. We hope that the new plots and statistics presented herein address the concern put forward by the reviewer.

      *Response Fig 1: A comparison of cell wise versus animal-wise analysis of synaptic physiology. Some cell to cell variability is hidden, and the reduction in numbers impacts the P values.*

      (A) PPR of multipolar Prox1 Control for 14 cells from 9 animals (n/N=14/9) under baseline conditions and with MSOP, cell-wise comparison p = 0.02 , t = 2.74 and (B) animal-wise comparisons (p = 0.04, t stat = 2.45). Statistics: paired t-test.

      (C) PPR of multipolar Prox1 KO cells (n/N=9/8) under baseline conditions and with MSOP, cell-wise comparison p = 0.2, t = 1.33 and (D) animal-wise comparisons (p = 0.2, t stat = 1.56). Statistics: paired t-test. Comparisons for PPR of bipolar Prox1 Control (n/N=8/8) and KO cells (n/N=9/9) did not change.

      (E) PPR for Prox1 control (n/N=18/11) and KO (n/N=13/11) bipolar VIP cells, cell-wise comparison p = 0.3, t = 1.1 and (F) animal-wise comparisons (p = 0.4, t stat = 0.93). Statistics: t-test.

      (G) PPR of Elfn1 Control (n/N=12/4) and Het (n/N=12/4) bipolar VIP cells, cell-wise comparison p = 0.3, t = 1.06 and (H) animal-wise comparisons (p = 0.4, t stat = 0.93)

      (I) PPR of Prox1 control (n/N=33/18) and KO (n/N=19/14) multipolar VIP cells, cell-wise comparison p = 0.03, t = 2.17. and (J) animal-wise comparisons (p = 0.07, t stat = 1.99).

      (K) PPR of Elfn1 Control (n/N=14/6) and Het (n/N=20/8) multipolar VIP cells, cell-wise comparison p = 0.008, t = 2.84 and (L) animal-wise comparisons (p = 0.007, t stat = 3.23).

      3) Clarify what are the parameters used to identify bipolar vs multipolar VIP cells. VIP cells comprise a wide variety of transcriptomic subtypes, and in the absence of using specific genetic markers for the different VIP subtypes, the authors should either include the reconstructions of all recorded cells or clarify if other methods were used.

      We thank the reviewer for this comment. The cell parameter criteria will be amended in the methods: “Cell type was classified as bipolar vs. multipolar based on cell body morphology (ovoid vs. round) and number and orientation of dendritic processes emanating from it (2 or 3 dendrites perpendicular to pia (for bipolar) vs. 3 or more processes in diverse orientations (for multipolar). In addition, the laminar localization of the two populations differs, with multipolar cells found primarily in the upper layer 2, while bipolar cells are found throughout layers 2 and 3. Initial determination of cell classification was made prior to patching fluorescent-labelled cells, but whenever possible this initial assessment was confirmed with post-hoc verification of biocytin filled cells.”

      Reference:

      Dolan, J., & Mitchell, K. J. (2013). Mutation of Elfn1 in Mice Causes Seizures and Hyperactivity. PLOS ONE, 8(11), e80491. Retrieved from https://doi.org/10.1371/journal.pone.0080491

      Haas, K. T., Compans, B., Letellier, M., Bartol, T. M., Grillo-Bosch, D., Sejnowski, T. J., … Hosy, E. (2018). Pre-post synaptic alignment through neuroligin-1 tunes synaptic transmission efficiency. ELife, 7, e31755. https://doi.org/10.7554/eLife.31755

      Horvath, P. M., Piazza, M. K., Monteggia, L. M., & Kavalali, E. T. (2020). Spontaneous and evoked neurotransmission are partially segregated at inhibitory synapses. ELife, 9, e52852. https://doi.org/10.7554/eLife.52852

      Stachniak, T. J., Sylwestrak, E. L., Scheiffele, P., Hall, B. J., & Ghosh, A. (2019). Elfn1-Induced Constitutive Activation of mGluR7 Determines Frequency-Dependent Recruitment of Somatostatin Interneurons. The Journal of Neuroscience, 39(23), 4461 LP – 4474. https://doi.org/10.1523/JNEUROSCI.2276-18.2019

      Tomioka, N. H., Yasuda, H., Miyamoto, H., Hatayama, M., Morimura, N., Matsumoto, Y., … Aruga, J. (2014). Elfn1 recruits presynaptic mGluR7 in trans and its loss results in seizures. Nature Communications. https://doi.org/10.1038/ncomms5501

      Udakis, M., Pedrosa, V., Chamberlain, S. E. L., Clopath, C., & Mellor, J. R. (2020). Interneuron-specific plasticity at parvalbumin and somatostatin inhibitory synapses onto CA1 pyramidal neurons shapes hippocampal output. Nature Communications, 11(1), 4395. https://doi.org/10.1038/s41467-020-18074-8

  9. Oct 2020