Effects of Valproic Acid Embryonic Exposure on Zebrafish: A Systematic Review and Meta-Analysis

Round 1

Reviewer 1 Report (Previous Reviewer 1)

Comments and Suggestions for Authors

Thank you to the authors for responding to all my comments. I accept the manuscript in its current form.

Reviewer 2 Report (Previous Reviewer 2)

Comments and Suggestions for Authors

No comments

Comments on the Quality of English Language

No comments

Reviewer 3 Report (Previous Reviewer 3)

Comments and Suggestions for Authors

The Authors considered my previous comments. Now the manuscript is suitable for publication. However, in my opinion the number of authors is too high.

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

 

The manuscript entitled “Effects of valproic acid embryonic exposure on zebrafish: a systematic review and meta-analysis” by Flores-Prieto et al. is a systematic review and meta-analysis summarizes the effects of embryonic exposure to valproic acid (VPA) in zebrafish over the last ten years. The authors conducted this review to elucidate the impact of VPA on development and molecular and epigenetic changes in zebrafish. As the authors emphasize, one of the main obstacles in comparing the results obtained under different protocols was methodological variability. Therefore, one of the key factors taken into account in the analysis were the duration of pharmacological exposure and the concentration of VPA used. The conducted meta-analysis provided evidence of the teratogenic effect of VPA in zebrafish. The most common side effects that occurred were: alterations in heart rhythm, neurological changes and epigenetic regulation. One of the main factors that should be given attention in the future is the standardization of methodologies that adapt and limit the adverse effects of VPA at an early stage of development.

On the whole, this review seems well conducted, methods are sound, data are convincing and the conclusions are in general supported by the data.

A few issues, which need to be addressed:

·         the abbreviation of GABA should be developed

·         the abbreviation of hpf should be developed

·         the abbreviation of  HDAC system should be developed

Author Response

Thank you for your valuable feedback. We have addressed the suggested points as follows:

• The abbreviation "GABA" has been developed as "gamma-aminobutyric acid."
• The abbreviation "hpf" has been developed as "hours post-fertilization."
• The abbreviation "HDAC system" has been developed as "histone deacetylase complex system."

We appreciate your attention to these details, which have helped us improve the clarity of the manuscript.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript explains the overall effects of valproic acid on the neurodevelopment in Zebrafish.However, manuscript suffers from lot of defects few of them are:

(A) Explanation of data: The authors have tried to explain effects on different stages of embryo development. However, in the whole manuscript, I dont see authors considering the stage specific effects of  VA and what are the factors that may or may not affect such stages. In the analysis, there is no mention of such factors and how they may affect such stage e.g. in blastula and gastrula there is cell number difference effect, gene expression etc.

(B) Poor discussion:In the discussion section of the manuscript, authors have discussed so many results together without linking one result with other and hence it is difficult to follow one discussion after another. 

Author Response

Thank you for your constructive feedback. We have made the following revisions based on your comments:

(A): We acknowledge the importance of considering stage-specific effects of valproic acid (VA) during embryo development. We have now expanded the analysis to discuss factors specific to the blastula and gastrula stages, such as cell number differences and gene expression. These additions clarify how VA might differentially impact each developmental stage.

(B): To improve the flow of the discussion section, we have restructured it to more clearly link each set of results, creating a logical progression from one point to the next. This revision enhances the readability and coherence of our findings.

We appreciate your detailed insights, which have greatly strengthened our manuscript.

Reviewer 3 Report

Comments and Suggestions for Authors

The topic of the work is interesting. Valproic acid is still widely used in pharmaceuticals. The manuscript refers to an important topic - the methodology of research on the teratogenicity of valproic acid on zebrafish model. The results obtained by Authors indicate the heterogeneity of the research protocols used.

The authors comprehensively described the methodology. They analyzed data from 39 articles from PubMed and Google Scholar. The results are presented in the form of 7 figures. Discussion and introduction properly written.

Objections:

-the quality of the figures (readability, resolution) should be improved

-number of authors - 11 - is far too many for the presented review paper (considering the methodology of this work). The editor should decide whether the number of authors is adequate, but I suggest that the authors should justify the participation of each Author.

Author Response

Thank you for your feedback. We have addressed your concerns as follows:

• Figure quality: The readability and resolution of the figures have been improved to ensure clarity and enhance the visual quality of the manuscript.

• Number of authors: We have reviewed the authorship and reduced the list by one. Additionally, we have enhanced the justification for each author’s participation, detailing their specific contributions to the work.

We appreciate your observations, which have helped us improve the overall quality and presentation of our manuscript.