Erythrocyte Respiratory Activity of the Mitochondrial Complex of the Black Sea Thornback Ray (Raja clavata L.) under the Influence of Certain Activators and Inhibitors In Vitro

Round 1

Reviewer 1 Report (Previous Reviewer 3)

Authors corrected most of the details I asked in my first review. Now the figures are understandable. I have only a few minor corrections to suggest:

keywords: correct to - cartilaginous fishes, respiratory rate

line 75: replace "Salmon" by "rainbow trout"

line 128: scientific name must be in italics

line 133: delete "an"

line 218: the Figures are 2-5

All numbers must have dots and not commas for the decimals. For example: 11,2 must be 11.2

Author Response

Dear Reviewer,

Thank you very much for your comments and precise corrections.

We have addressed it in the manuscript and explained below in this document for each valuable question or remark you have asked for.

Please let me know if you need us to provide any additional information,

 

Many thanks,

Best regards,

 

Alla Silkina

[email protected]

Author Response File: Author Response.pdf

Reviewer 2 Report (New Reviewer)

The authors that examined the role of mitochondrial complex of nuclear erythrocytes

In cartilaginous fish thornback ray (Raja clavata L.) during encountered the climatic fluctuations on the energy basis of blood cells. They found that some activators such as glutamate, maleate and succinate to the erythrocyte suspension caused a 2 - 6-fold increase in the respiratory activity of thornback ray erythrocytes. In contrast, some inhibitors such as Protonophore – dinitrophenol, can caused inhibition of the activity of mitochondrial respiration. Final, they summary that the activation of "respiration" of erythrocytes in fish, which point out the potential capabilities of cold-blooded and warm-blooded vertebrates have a very similar approach via the inner membrane to support the energy potential of the mitochondria of the cell.

This topic is interesting and some comments as following before consider for publish.

Comments:

1.     Please provide a summary carton for outline the results and discovery.

 

2.     The title should be more precise and significant on erythrocyte function.

 

3.     What are activators and inhibitors should be defined in the Introduction section. And what does that means in in the respiratory activity of thornback ray erythrocytes in mitochondria.

 

4.     In Figure 1, figure legend should be enhanced and more explaining them.

 

5.     Please if it possible should be checked in the ROS generation for understanding the oxidative stress for the treatment.

 

6.     Please explained the adaptive strategy of the erythrocyte mitochondrial complex in the thornback ray (Raja clavata L.)?

Author Response

Dear Reviewer,

Thank you very much for your comments and precise corrections.

We have addressed it in the manuscript and explained below in this document for each valuable question or remark you have asked for.

Please let me know if you need us to provide any additional information,

 

Many thanks,

Best regards,

 

Alla Silkina

[email protected]

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report (New Reviewer)

It is a better that can consider for publish.

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

The information in this paper is a novel and very interesting contribution the physiology of Black Sea thornback ray. I only recommend including some details in the statistical methods and results which can improve the clarity of the information:

 

In the experimental design lines 100 to 110, there is no information about sample size and variation between individuals. How many fish were sampled for blood extraction? and how variable is the oxygen consumption between individuals, sexes, and age on the experiments? Authors should include this information.

Reviewer 2 Report

This manuscript was not clear to me. It appears that the different chemicals were added sequentially (according to the presented figures). From the figures it is (to me) very difficult to evaluate, what their effects on respiratory rate are. It seems that the number below the curve close to the arrow represent the value for the oxygen consumption for that chemical. However, one problem to me is that it seems that the figures and the measurements give only one set of erythrocytes. Indeed, the authors say that the figures give "typical" experiment. Yet, afterwards they say that the four experiments were compared using t-test. Where? Further, the rationale of the work is not clear to me. I also question, if N=4 would be adequate even if the mean values for the measurements were given somewhere.

Reviewer 3 Report

keywords: do not use words that are in the title

The introduction of this manuscript is clear, focused to the subject of study. line 72 actual name: Oncorhynchus mykiss

line 78: number of citation?

lines 86-90: reference for these statements?

line 102: "from" and not "form"

line 103: replace "nexty chapter" by "section 2.2"

line 124: rays were kept for how many days?

line 125: were the rays anesthetized?

Part of the material and methods was clear to me, but how the inhibitors were used was not clear, and results also were difficult to understand. I also looked to a published article of the authors, in which the figure captions are more complete, but still the way they place the data in the graphics is not clear to me.

lines 139-144: why were the inhibitors added sequentially to the same preparation? The concentrations are not in the figure captions, or at least not in a way that I could understand.

lines 160-163: where is this sixfold increase in fig. 2?

lines 165-166: what is "sea fox erythrocyte suspension"? All abbreviations in the figure must be explained in the caption. Decimals must be with dots and not commas. What does the numbers below the curve represent?

What was the concentration of the mixture of salts of glutamic and maleic acids (G + M)? 

line 250: neither reference number  9 nor the article of Vatin and Danilenko, 2020 contain this information. Is is reference number 12.

Discussion seems to be appropriate, but as I had a lot of difficulty to understand results, I could not evaluate properly.

Reviewer 4 Report

The main idea of the current study seems so good. The author had been made a great effort in this research paper. the manuscript could be accepted for publication after considered some minor corrections:

The English editing need to revised by native language speaker.

L 121 need to specify in mentioned the exact location (latitude degrees).

L 124 what about the anesthesia method before blood collection?

L 132 need reference for this procedure.

L 141 need reference for this procedure.

L 147 the company information should be addressed not website?

L 148 The collected data need to be examined for their homogeneity and normal disruption prior to data analysis.

Conclusion part should be shortened and concise, it is so long.