PGENETICS-D-25-01073

Octopamine signaling from clock neurons plays dual roles in Drosophila long-term memory

PLOS Genetics

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Giovanni Bosco, Ph.D.

Section Editor

PLOS Genetics

Giovanni Bosco

Section Editor

PLOS Genetics

Aimée Dudley

Editor-in-Chief

PLOS Genetics

Anne Goriely

Editor-in-Chief

PLOS Genetics

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Authors:

Please note that one review is uploaded as an attachment.

Reviewer #1: The authors described the role of octopamine from two distinct populations of clock neurons on long-term memory (LTM), which is a non-clock function in Drosphila melanogaster. They found PDF-neurons, which includes large and small ventral lateral neurons (sLNvs and lLNvs) functions in the consolidation of LTM, while dorsal lateral neurons (LNd) functions in the maintenance of LTM. Although these neurons functions in circadian rhythm regulation and octopamine in sleep regulations, alteration of octopamine signals from these neurons had no effects of circadian rhythm and sleep. The experiments were well designed and have been performed under contemporary standards, and the results well supported their conclusion. This study described non-clock function of clock neurons, which is novel and brought significant insights into this field with its clarification of the molecular mechanism.

I have a couple of minor comments as follows, before it can be accepted;

1. It is preferable to show the RNAi specificity. The authors used only 1 RNAi line in PDF neuron experiments, which can be improved if they used additional lines, like LNd. In addition, rescue experiments will also strengthen the conclusion.

2. As for R78G02 experiments, the authors guessed the involvement of non-clock neurons. This conclusion solely depends on the results of R54D11 and LNd-split GAL4 experiments. However, the efficiencies of RNAi by different GAL4 are different, even they are expressed in the same cells. Therefore, the results can be interpreted in another way. It is also preferable to use more specific GAL4 drivers, which show positive effects in memory functions.

3. The authors described that the baseline of the courtship index (naïve CI) were different between LNd-split-GAL4/UAS-Tbh RNAi “1 and Pdf-GAL4/UAS-Tbh RNAi #1. Since memory index of LTM is affected by the native CI, they should mention how this difference may affect the results in the discussion.

4. The authors concluded Tbh knockdown did not affect the circadian rhythm and sleep. But in the supplemental figures (S2, S6), there are minor changes in a couple of parameters. Thus, they should rephrase, maybe “There were minor effects which are not likely to affect LTM.”

Reviewer #2: uploaded

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Large-scale datasets should be made available via a public repository as described in the PLOS Genetics data availability policy , and numerical data that underlies graphs or summary statistics should be provided in spreadsheet form as supporting information.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Sakei review.docx

Dear Dr Sakai,

We are pleased to inform you that your manuscript entitled "Octopamine signaling from clock neurons plays dual roles in Drosophila long-term memory" has been editorially accepted for publication in PLOS Genetics. Congratulations!

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Yours sincerely,

Giovanni Bosco, Ph.D.

Section Editor

PLOS Genetics

Giovanni Bosco

Section Editor

PLOS Genetics

Aimée Dudley

Editor-in-Chief

PLOS Genetics

Anne Goriely

Editor-in-Chief

PLOS Genetics

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Comments from the reviewers (if applicable):

Reviewer's Responses to Questions

Comments to the Authors:

Please note here if the review is uploaded as an attachment.

Reviewer #1: The authors responded to all my concerns satisfactory, and I have no more comments.

Reviewer #2: I am satisfied with the additions and changes that have been made.

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Have all data underlying the figures and results presented in the manuscript been provided?

Large-scale datasets should be made available via a public repository as described in the PLOS Genetics data availability policy , and numerical data that underlies graphs or summary statistics should be provided in spreadsheet form as supporting information.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes: Kazuhiko Kume

Reviewer #2: No

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