Comparative Transcriptome Analysis of the Skin and the Peritoneal Wall Layer of Triplophysa stenura Distributed in High Elevations

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

I believe that it is an interesting work and that it achieves comparative transcriptomics results in relation the differential expression in the analysis of the skin and the peritoneal wall layer of Triplophysa stenura distributed in high-elevation.

I consider that the authors have performed a good experimental design; however, the authors must to explain why to select to different skin samples in the experimental design? What is the underlying hypothesis?

The authors have recovered the transcriptomic profile of DEGs in a good Discussion about the importance of different set of genes that could be explaining the different environment adaptation, and that could be followed in further studies. I have only suggested small changes in particular in text edition.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

No comments

Author Response

Dear editor and reviewers,

We thank you all for your effort to improve the quality of our manuscript. Reviewers’ valuable comments and suggestions were welcome and helpful for us to revise our manuscript. Our detailed response was attached as a file.

Thank you!

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript addresses a complex topic, such as the expression of unigenes in different areas of the skin of Triplophysa stenura. I have been pleasantly surprised by the reading of the manuscript. However, I believe that there is an overuse of very short paragraphs, especially in the introduction and methods sections, making the reading somewhat incoherent. Here are the comments from my review:

55. The citation should be indicated at the end of the sentence.

73-75. This sentence is confusing and should be rephrased.

87. It would be advisable to clarify why only 3 fish were decided to be used; it seems like a small number.

90. What happened to the fish? While a dose of 200 mg/L of MS-222 is an anesthetic concentration, were the fish subsequently euthanized? Was the MS-222 buffered with bicarbonate, for example, to attenuate its acidic nature?

142. The left side is missing from the equal sign (=) in the equation.

163-168. There are several spelling errors with units in this paragraph. Check the bibliographic citation at the end of it.

193. Why this expression? "According to your message."

Figures 5 and 6 are difficult to read.

245-253. This paragraph is somewhat confusing. It should be rewritten.

255. The text you provided describes the results of a GO annotation analysis?

256. Double space.

The same issue occurs with figures 5 and 6.

317-319. Correctly cite the URLs.

I know it is not a mandatory requirement, but adding DOIs to the references greatly facilitates their retrieval.

Comments on the Quality of English Language

I have no comments.

Author Response

Dear editor and reviewers,

We thank you all for your effort to improve the quality of our manuscript. Reviewers’ valuable comments and suggestions were welcome and helpful for us to revise our manuscript. Our detailed response was attached as a file.

Thank you!

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The Decision on the manuscript with ID (biology-2737399-peer-review-v1) is “Major Revisions”. Whilst the study was so interesting; however, the manuscript needs several revisions prior considered for publication in Biology. The authors should prepare a point-by-point response to the comments and points raised by the anonymous reviewer.

Major comments: -

I have 5 main questions.

Q1. Line 87: Did the authors find that “3 fresh and healthy fish” was enough to confirm the validity and replication of your results? I see that these three fish were not enough at all for replication of the results. Besides, what is the number of samples taken (n= ??)? Would these samplings be enough for appropriate statistical analysis?

Q2. Line 97: As I said above, the number of samplings (skin and peritoneum) should be clearly described in both the Material and Method sections as well as in the results under the figures.

Q3. Line 101: You described that gel electrophoresis was carried out to confirm the quantity and quality of the total isolated RNA. Why did you not provide these results even in the supplementary files?

Q4.  Line 167: Add a suitable reference for the used methodology.

Q5. Line 168: (Ye et al., 2006) – This reference was not found in the reference section.

Q6. For all figures, the authors should add the number of samplings used to conduct the analysis.

Q7. Figures 16 and 17: You should define the different colors to refer to the dorsal skin or to the peritoneal wall layer.

Minor comments: -

Line 25: T. stenura

Line 90: Add company name, City, and Country. Add a suitable reference for the used anesthetic dose. Did you use buffered MS222 or use it as it is? What is the induction time obtained?

Line 93: Add the ethical approval code.

Line 470: T. stenura

Lines 344-345: Figure 16: the dorsal skin and the peritoneal wall layer.

Comments on the Quality of English Language

Moderate editing of English language required

Author Response

Dear editor and reviewers,

We thank you all for your effort to improve the quality of our manuscript. Reviewers’ valuable comments and suggestions were welcome and helpful for us to revise our manuscript. Our detailed response was attached as a file.

Thank you!

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

The authors properly revised the manuscript. However, the manuscript needs English Editing and proof reading.

Comments on the Quality of English Language

Extensive editing of English language required