PONE-D-22-30720

Multivariate Analysis of Variegated Expression in Neurons: a strategy for unbiased localization of gene function to candidate brain regions in larval zebrafish

PLOS ONE

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Reviewers' comments:

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Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: This work describes the use of a protocol developed by the authors to identify a zebrafish brain region in which the CaSR gene acts to control response to an acoustic stimulus (reorientation versus escape). The technique, MAVEN, is an unbiased correlative approach that relies on variability of expression of Gal4 driver lines in zebrafish brains. In other words, each transgenic larva expresses the driver in a different set of brain regions. Sorting animals by behavioral phenotype and then correlating regions of brain expression is the basic idea, but a thorough set of statistical and computational analyses is employed to derive a correlation of expression to behavioral phenotype.

This work is very important because, especially in vertebrate behavior, connecting phenotype to brain region is, as the authors say, a bottleneck. This technique turns a disadvantage (variegated transgene expression) into an advantage to make these phenotype-expression region correlations that can relatively quickly identify the brain region.

The work identified the DCR6 (rhombomere 6) region of the brain as correlating with rescue. Known DCR6 Gal4 expression lines were then directly used to confirm the importance of CaSR expression in that region.

This work should definitely be published so that the greater community can begin to use it to determine its broad utility. The protocol has already been published on protocols.oi.

Comment:

The “gain-of-function” and “loss-of-function” approaches could be better explained when first mentioned. “Loss-of-function”, I assumed, means rescue of a mutant, but it was not stated exactly what “gain-of-function” means (e.g. overexpression?). There seems to be some intermingling of these concepts when the results are described.

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Reviewer #1: No

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Multivariate Analysis of Variegated Expression in Neurons: a strategy for unbiased localization of gene function to candidate brain regions in larval zebrafish

PONE-D-22-30720R1

Dear Dr. Granato,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Bruno Giros, Ph.D.

McGill University

Academic Editor

PLOS ONE

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