Maternal Pregestational Diabetes Contributes to Neural Tube Defects in Mouse Fetuses Through H4K5ac-Mediated Regulation of Focal Adhesion Pathway

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript titled “Maternal Pregestational Diabetes Contributes to Neural Tube Defects through H4K5ac-mediated Regulation of the Focal Adhesion Pathway” presents a well-designed study. The authors investigated changes in protein and gene expression levels in E9.5 mouse brain tissue. The KEGG and GO analyses support the proposed hypothesis and provide valuable mechanistic insights into the relationship between maternal pregestational diabetes and neural tube defects.

Overall, the data appear reliable, and the figures and tables are of good quality. In addition, the references are current and appropriate. However, several issues should be addressed before the manuscript is considered for publication.

1. The experimental design could be improved. The authors pooled three samples and performed duplicate analyses instead of triplicates. Since MALDI-TOF MS is a highly sensitive technique for protein detection, it may have been possible to divide the 200 μg protein sample into three groups to perform triplicate analyses, which would strengthen the statistical reliability of the data.
2. Please specify how much total protein was used for the proteomic analysis.
3. Please indicate how much mRNA was used for cDNA synthesis.
4. Abbreviations should be defined at their first appearance in the manuscript. Examples include:
   • H4K5ac (line 15)
   • FVB (line 17)
   • TMT (line 20)
   • Gsk3b and Mapk9 (line 27)
   • ATRA and MALDI (lines 45 and 52)
5. Please check the formatting of Gsk3b in lines 32 and 36 to ensure consistency with journal style requirements.
6. In lines 111–112, the group names should be clearly specified to improve readability and help readers better understand the figures and results.
7. In the Figure 2 caption, please explain why certain proteins are highlighted with red boxes. Similar clarification should also be added to all other figures containing red boxes.
8. E9.5 should be embryonic  even though it is same meaning at gestation. 

Author Response

Thank you for the reviewer’s comments. Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Manuscript ID: genes-4323503

Type of manuscript: Article

Title: Maternal Pregestational Diabetes Contributes to Neural Tube Defects through H4K5ac-mediated Regulation of Focal Adhesion Pathway.

 

This study aimed to investigate the potential mechanisms of maternal pregestational diabetes induced neural tube defects (NTDs) by integrating proteomic data and H4K5ac ChIP-seq data from the mouse model. 

 

Comments and Suggestions for Authors:

 

The manuscript is an interesting article but requires some consideration.

 

Throughout the manuscript, acronyms are used incorrectly (TMT, RT-PCR, FVB, NTDs, etc), sometimes before they have been previously described in the first time that they have been used or re-describing them after having done so previously. This should be corrected.

Including a section of Abbreviations used in the manuscript would be very convenient.

 

Title.

The title should include that an animal experimentation model is being studied.

 

Abstract.

It should be included that the diabetic mouse model was established in 6 female FVB mice.

 

1. Introduction.

The Introduction section should end with the objective of the study.

The conclusions reached in this section (lines 77-81) should be reserved for a Conclusions section.

 

2. Materials and Methods.

The dates on which the study was conducted and the research center where the study was conducted should be included.

Authorization, including the approved protocol, from the Animal Experimentation Ethics Committee should be included.

An assessment of the sample size calculation should be provided and discussed a priori to demonstrate the stated objectives.

Describe the efforts made to address any sources of bias in the study.

 

3. Results.

Lines 250-256. This paragraph would be better included in the Discussion section.

 

4. Discussion

The study's limitations should be discussed more extensively, such as the consideration of other factors that were not considered in the study.

 

5. Conclusions

A Conclusions section is missing. It should be included as is the norm for the Genes journal.

 

References:

References must be corrected according to the norms of the Genes journal.

 

Author Response

Thank you for the reviewer’s comments. Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

Round 2.

In the new V2 manuscript, the authors have made changes based on the recommendation of referee that improve its presentation. The reviewer thanks the authors for their effort in facilitating the assessment of the changes made.

Accept after minor revision (text editing).