Construction of a Zebrafish Model of Cardiac Hypertrophy Caused by ATIC Gene Deletion and Preliminary Exploration of Aerobic Exercise Improvement

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The study is interesting showing the hypertrophic cardiomyopathy symptoms by silencing atic-/-zebrafish heart and later effect of aerobic exercise was noticed on the cardiomyopathy symptoms. I have some concerns that need to be addressed:

1. Author should describe the rationale behind the selection of zebrafish (vertebrate) using as model organism. Explain the advantages and limitations of zebrafish on the other model system such as mammalian (mouse and rats). Also, describe how the study out outcome can be translated to humans.  
2. line 23, EMC receptor, write the full name of abbreviation while appearing first in the text.
3. line 38 delete the word “is converted”.
4. For demonstrations of the exercise experiment it would be better if author could provide the videos as supplementary files or at least author could make ray diagram and insert it with main text or as supplementary figure.
5. In section 2.4 what is HE staining, write the proper name H&E staining. Similarly in section 2.5 MASSON staining.
6. Section 2.7. initiates with capital alphabets.
7. What is 5.-month (line 142).
8. Is it 35- month (line 152)? Correct it.
9. Figure 1. initiates with capital alphabets (line 190).
10. Correct grammatical and typo error entire MS such as tic gene (line 190, 202, 203, 238, 239 etc.).
11. The histological representation in Figure 2 is not clear. Either enhance the resolution of the images to show the prominent histological changes or provide some zoomed section and place as supplementary file.
12. Follow the standard guidelines suggested by the journal for the references.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

 

1. Referencing numbering issue at line 52 onward (check references and numbering from ref 8 onwards).
2. Section heading: 2.7 Transcriptome.
3. Replace 5. -month → 5-month.
4.  Replace statistic → Statistic.
5. 6   Line 200: replace tic → atic.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

In the manuscript named “Construction of Zebra Model of Cardiac Hypertrophy Caused by ATIC Gene Deletion and Preliminary Exploration of Aerobic Exercise's Improvement”, authors had constructed CRISPR/Cas9 gene editing zebrafish, and they had performed cell straining, RNA-seq and qRT-PCR to investigate genetic regulation pathways in hypertrophic cardiomyopathy. Their findings would be helpful for human health in future, but the manuscript was poor for publishing at this present form.

(1) In introduction section, authors needed to elaborate on the relationship between HCM, ATIC, and exercise with detailed explanations.

(2) In Figure 7, the Apelin signaling pathway was significant in GSEA analysis, but it was not in figure 5D.

(3) Authors had used “Beijing Nuohe Zhiyuan Biotechnology Co., Ltd. Official website” line 145, but the Nuohe was translated as “Novogene”, please access the website to check it. In addition, the RNA-seq analysis was almost not described in method section. For example, how to map, how to calculate genes expressions, how identify DEGs, and how to perform GSEA analysis.

(4) The RNA-seq would be released on public database, not requested by readers.

(5) Authors had generated two lines (gene editing lines), but these sequences information were inconsistent with the sequencing electropherograms, see figure 1, and corrections by the authors were required.

(6) From the CRISPR/CAS9 lines, authors had displayed with no expressions in gene editing lines, see figure 1C, authors needed to confirm it, for example, authors could access these results from RNA-seq mapping results.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript is substantially modified based on the asked questions. I endorse the publication of the article. 

Author Response

Thank you for your review comments. Wishing you a smooth work and a happy day every day！

Reviewer 3 Report

Comments and Suggestions for Authors

Thanks for authors’ works, the problems were still present in revision. For example, “Nuohe Zhiyuan” was still present in manuscript, the bioinformatics was still too simple, why did they use “edgeR”, how did F-test perform? In addition, the data was submitted to NCBI, but accession number or review link was not present, and authors still declared “The raw data behind the conclusions of this study are available from the corresponding author upon reasonable request.”, please check them. Good luck.

Author Response

Thank you for your review comments. Wishing you a smooth work and a happy day! (Please see attachment)

Author Response File: Author Response.docx