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The authors have identified multiple types of "Janu" neurons with specific anatomical and neurochemical properties (e.g., GABA, AstA). These neurons all control thirst seeking, and also other unique (but related) behaviors. The Janu-AstA neuron is argued to synapse with and signal to NPF neurons. Focusing in on the key data in support of these main claims, I am only mildly convinced. The strongest phenotypes they observe are always with the broad drivers (e.g. Durstig-Gal4, which has widespread expression through the brain), but once they home in on a split-Gal4 combination with very selective labeling, the phenotype effects of activation or suppression are very small (e.g. Figure 2J-K or the new optogenetic activation experiments in Figure 5E). For some experiments, they revert to using the broader driver, such as the GRASP with NPF neurons using R65D05-Gal4. Unfortunately, this Gal4 line is expressed in a mass of neurons in this region of the brain (Figure 2S1A), so it's not clear to me that a positive GRASP signal really reflects synaptic contact between Janu-AstA neurons (somewhere within the R65D05 pattern) and NPF neurons. As mentioned in the previous round of reviews, the baseline phenotypes of controls is variable (e.g. in Figure 3I and Figure 3S1A VGAT RNAi experiments, there are many fewer flies on H2O compared to those with Gad1 RNAi in Figure 3J). The authors somewhat dismissed this concern by arguing that this may reflect variable environmental conditions and water deprivation treatments of flies of experiments on different days (and they only compared assays run in parallel), but this remains a concern for me. The n of several assays seems rather low (6-7), so it's not clear if they did all experiments of one type on a single day. In any case, questions linger as to how robust some of their results are.
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As mentioned in the previous round of reviews, the baseline phenotypes of controls is variable (e.g. in Figure 3I and Figure 3S1A VGAT RNAi experiments, there are many fewer flies on H2O compared to those with Gad1 RNAi in Figure 3J). The authors somewhat dismissed this concern by arguing that this may reflect variable environmental conditions and water deprivation treatments of flies of experiments on different days (and they only compared assays run in parallel), but this remains a concern for me.
Zip code 66286 is located in Shawnee, Kansas.The 2023 sales tax rate in Shawnee is 9.6%, and consists of 6.5% Kansas state sales tax, 1.48% Johnson County sales tax and 1.63% Shawnee city tax.
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