two cervical spinal cord injury (SCI) models: C2 hemisection (C2HS) and C3 hemicontusion (C3HC)
Perhaps this comparison has already been done, but did you consider also including a control group of mice that received a superficial surgery (skin and muscles retracted) but no cervical spinal cord injury? I'd be curious whether/how much the localized tissue inflammation had downstream effects on the cervial spinal cord.
Finally, MEA recordings revealed functional responses to histamine and hIL-31
I'm not sure if these data are available, but is it possible to compare the increase in mean firing rate with HA or IL-31 treatment in iPruriceptors vs primary human pruriceptors?
Notably, the choice of protocol (1-step vs. 2-step) had a greater impact on differentiation outcomes than the identity of the overexpressed TF as shown by the principal component analysis
Wow, very interesting. Why do you think this is true?
HRH1 receptor
H4 too, right?
Clever approach! I wonder though if target-protein co-localization on the membrane is a good model for target interactions with free proteins?
Lovely structure figures!
could directly compete with the MRGPRX2 agonist 48/80 in its ability to activate the G protein Gi, an established signaling output of MRGPRX2
I wonder if this might be a better success metric than binding, if the goal is to predict nanobodies that could be developed into therapeutics?
This receptor was chosen because its shallow binding pocket suggests a more forgiving thermodynamic landscape relative to other GPCRs
MrgprX2 is highly promiscuous (binds a huge list of basic secretagogues). Do you think this makes it easier to predict a functional nanobody? Maybe this is exactly what you're getting at here
TRPA1 is a cold sensor in mouse, rat and other mammals (Kwan et al., 2006; Story et al., 2003; Zhang et al., 2022), and a heat sensor in amphibians and reptiles
Interesting! Are "cold" and "hot" considered different stimuli?
I don't understand what you mean by "found in the AD map"?
Hi! I'm not sure what this means. Could you give a one-sentence explainer of "systems biology standards" for the non-systems biologists?
A prevalent error in the numerous predictions we examined involved the occurrence of brief sequences (up to 5 frames) characterized by false-negatives or-false positives.
Did the prevalence of these errors increase with particular behaviors?
Frames depicting paw contact before (top panel) and during flinching, overlaid with a brightness analysis (threshold > 150) (bottom panel)
Great figures! I wonder if it would be possible to include a short video?
Nc – indicates Negative contro
What treatment did the negative control rats receive? Many symptoms of AD, including itch, can be largely resolved just by applying moisture to the skin. It might be informative to run a vehicle-only control, to separate the effect of the compounds from the effect of the oil carrier.
Figure 1
I see 5 treatments in the legend but only 4 bars. The negative control appears to be missing? Or did those mice not scratch at all?
Allergen-treated animals displayed elevation in non-evoked pain as measured by the Mouse Grimace Scale (MGS
Why do you think you saw an increase in grimacing in the control group throughout the time course?
a significant reduction in the lysosomal mass as evidenced by lower lysosomal mass index: −1.823 [−2.617 to −1.028] p=0.0053, but no differences in lysosomal acidification represented as lysosomal acidification index
Did you expect to see a change in the acidification in control vs cytokine-treated?
resulted in limiting inflammation
How would degranulation limit inflammation? The MCs are releasing pro-inflammatory cytokines, as you pointed out.
Additionally, mast cells (MCs) were increased in the lungs of macaques with PTB
Consider including a quantification of the increase reported by Esaulova et al? It's always difficult to evaluate the importance of a finding like this without some appreciation of the magnitude of the change.
It is unclear whether this is a systematic, replicable difference denoting a further increase in maturity at D70, or whether it simply reflects the variability expected from small molecule iSN differentiations
Is there another experiment you think could answer this question?
are sufficient to suppress itch
Another reader left a similar comment above, but I wonder if you've considered trying other pruritogens besides chloroquine? Chloroquine is often used to model non-histaminergic itch, but (of course) histaminergic itch is also important, and accounts for a large proportion of itch signaling.
changes in the jejunum
Wow, is this reaction typical for mechanical stress at the level of the skin?
Figure 2.
Hi there! Looks like the figure images/links are broken
Did you consider using structure as a search parameter, in addition to sequence?
Thus, we infer that the decades’ prior use of translationally non-relevant, high-doses of morphine in rodents was likely selected and optimized to modify reflexive detection metrics that are not reflective of the human pain experience and do not assess the effect of morphine on pain affective-evaluative dimensions
Interesting and important inference! Have you considered using your setup to do dose response tests for morphine?
Our resulting models outperform their purely supervised counterparts across a range of metrics and datasets, providing more reliable predictions for downstream analyses
Fantastic! I'd love to see some specific comparisons
Interestingly, the most common syllables used by the workers were freezing
Your analysis of the behavioral syllables is fascinating! In regard to the freezing, have you considered whether the mole rats were responding to specific stimuli in the room? Was an experimenter present during filming?
olive oil (vehicle)
Did you notice any inflammatory or adjuvant effect from the olive oil?
5.7% in individuals with LD vs. 5.0% in individuals without LD
5.7% vs. 5% does not seem like strong evidence of an association... is this a significant difference?
hair loss, skin flaking, hyperpigmentation, hemorrhaging, and ulceration
Are these symptoms associated with AD in Lyme disease patients?
tends to branch out of senescence
Interesting paper so far! I'm not sure what this phrase means, though... Do you mean AD in older patients is more severe?
Collectively, these findings provided evidence that tick EVs functionally regulate DETCs in the murine skin
Beautifully written paper!
Regarding the EVs' regulation of DETCs: how long does the effect last? If you allowed the ticks to feed for 3 days, then removed them and waited another 48 hours before taking the biopsies... do you expect the DETCs would still show impaired rounding?
SC as a competitive inhibitor of TRPA1 and TRPV1
The docking models are interesting, but do you have any empirical data that indicate SC binds TRPA1/TRPV1?
DEM group, CPZ group, and all SC
Very interesting work! Wondering if you could explain the rationale for including DEM and CPZ groups in this experiment. I assume these are typical treatments for ACD?
n other words, there appears to be a subset of GRPR neurons that are activated by multiple itch-inducing compounds, which we refer to here as GRPR itch neurons
Have you considered testing an algogen alongside pruritogens to confirm that these neurons are specific to itch?
This test was conducted in the morning
Hi there! Just wondering why you specifically conducted this test in the morning? Have you noticed changes at other times of day?
maximum length of 10s
This sounds like an incredible setup! Is it possible to collect longer videos?
mice developed significant spontaneous scratching that increased over time
Wow, the results look profound. Just curious about the onset of the scratching behavior. I'm looking at the time course data in Sup Fig 1b... why did you start measuring scratching at 7 weeks? Is that when you started to notice scratching?
facial and nape paw swipes
Wow, what an intense response! I'm a little confused about the pain response data, though. Swipes are often considered an indication of pain, so I would interpret the behavioral data as evidence of increased pain in the Ddx3x(BRN3A) mice. But if I'm understanding correctly, the same mice showed a reduction in pain responses according to your hot plate and capsaicin injection experiments. Did I get that right? What's your thinking on this?
reduced DDX3X levels in cultured mouse DRG neurons
Looks like less of an effect (or maybe a slower effect?) in DRGs... wondering what you make of that?
(Figure 1A)
I'm confused about the labels in the figure. What does "IgE+Ag" refer to?
we expected that BMMCs are as sensitive as natural MCs in the body to various stimulation and kaempferol
I would be curious to hear more about your reasoning, here. Why do you think BMMCs are more sensitive to stimulation than MC cell lines?
The two camera feeds
Maybe I missed it elsewhere in the text, but could you provide detail about the cameras and capture settings?
Various behaviors were summarized in a behavioral profile
Did you validate these behavioral data with manual analysis of the videos?
(1) nose, (2) right ear, (3) left ear, (4) head, (5) neck, (6) center, (7) hip, (8) tail base, (9) right front paw, (10) left front paw, (11) right rear paw, (12) left rear paw
Wow, great! Did DLC reliably call all of these positions in most frames?
A more large-scale analysis must be performed to determine optimal spacer design criteria, and to test how different factors (melting temperature, GC-content, target site availability) influence KD efficiency
Agree! Seems like these are critical steps for establishing Csm-mediated RNA KD as robust and specific technique. Do you have plans for further characterization?
We varied the GFP-targeting spacer length from 24-48 nt in increments of 4 and assayed GFP KD
You measured GFP fluorescence, but did you also measure RNA levels directly? I'm curious how KD efficiency of targets like MALAT1, NEAT1 or TARDBP would change with spacer length.
The structure alignment shows that three agonists bind to different regions of the orthosteric pocket in three receptors, indicating distinct recognition mechanisms
Interesting! Is it possible to say anything about binding affinity for these ligands, or how they compare in activity assays?
The structure alignment shows that three agonists bind to different regions of the orthosteric pocket in three receptors, indicating distinct recognition mechanisms
Interesting! Is it possible to say anything about binding affinity for these ligands, or how they compare in activity assays?
We varied the GFP-targeting spacer length from 24-48 nt in increments of 4 and assayed GFP KD
You measured GFP fluorescence, but did you also measure RNA levels directly? I'm curious how KD efficiency of targets like MALAT1, NEAT1 or TARDBP would change with spacer length.
A more large-scale analysis must be performed to determine optimal spacer design criteria, and to test how different factors (melting temperature, GC-content, target site availability) influence KD efficiency
Agree! Seems like these are critical steps for establishing Csm-mediated RNA KD as robust and specific technique. Do you have plans for further characterization?