Mitochondrial Metabolism Characterization of Four Different Fish Cell Lines
Round 1
Reviewer 1 Report
Keywords: please avoid the use of words already reported in the Title. Try to substitute them with other related ones, to give more soundness to your manuscript.
Introduction: the role of mitochondrial metabolism in fishes was almost missed in the present form. Please, add some information about this topic, to better introduce and justify the use of these four cell lines of fishes and the related species, to enhance the relapses of your manuscript.
Materials and methods:
line 85, please specify when you used the Bonferroni test or the Game Howell test, on the basis of which detailed parameters, to give clear information to the readers.
Result and discussion (please address the name of the section)
line 98, please avoid the use of ...
lines 94-105: this period could be used to fix my previous comment about the Introduction section. Indeed, this is not a discussion but just an introduction of what I mean above, the role of the chosen cell lines in fish studies, also related to the species. I suggest you move this period to the Introduction section and leave a remainder sentence in the introductory part of the Discussion.
References: please double-check the reference's style and italicize the scientific names.
Best regards
Author Response
Keywords: please avoid the use of words already reported in the Title. Try to substitute them with other related ones, to give more soundness to your manuscript.
-Done.
Introduction: the role of mitochondrial metabolism in fishes was almost missed in the present form. Please, add some information about this topic, to better introduce and justify the use of these four cell lines of fishes and the related species, to enhance the relapses of your manuscript.
-Added.
Materials and methods:
line 85, please specify when you used the Bonferroni test or the Game Howell test, on the basis of which detailed parameters, to give clear information to the readers.
-Done.
Result and discussion (please address the name of the section)
- Both sections are together which is an option according to the author's guidelines.
line 98, please avoid the use of ...
-Done.
lines 94-105: this period could be used to fix my previous comment about the Introduction section. Indeed, this is not a discussion but just an introduction of what I mean above, the role of the chosen cell lines in fish studies, also related to the species. I suggest you move this period to the Introduction section and leave a remainder sentence in the introductory part of the Discussion.
- Done.
References: Please double-check the reference's style and italicize the scientific names.
-Done.
Reviewer 2 Report
General Comments
The purpose of this communication is to assess the mitochondrial status and plasticity of four fish cell lines, but this communication may not provide a good opportunity to enlighten other researchers. Frankly speaking, I am relatively neutral or even negative about the quality of this manuscript. In this study, mitochondrial metabolism was investigated in four cell lines of different species and different tissue origins（2 brains, 1 hepatocellular carcinoma, 1 fibroblast）. Without involving quantitative analysis, the reader should imagine that the mitochondrial metabolic patterns and levels of these cell lines of different tissue origins should be different.
Details:
1. Line 24, “REDOX” may be changed to lower case.
2. “Materials and Methods”. These cell lines required different culture temperatures and systems. Could the experimental design in this study distinguish whether the differences in mitochondrial metabolism were caused by the culture system or cell line differences?
3. “Results & discussion”. The purpose of comparing inherently different cell lines is not clearly communicated to the reader. It is not enlightening to speak of difference as difference. In addition, the use of four cell lines to test the differences in mitochondrial metabolism to guide peer researchers to select suitable cell lines for different experimental purposes (there are about more than 900 fish cell lines) seems to assume that most peers do not know the different mitochondrial metabolic characteristics of different types of cell lines, which may be inconsistent with the reality.
Author Response
- Line 24, “REDOX” may be changed to lowercase.
-Done.
- “Materials and Methods”. These cell lines required different culture temperatures and systems. Could the experimental design in this study distinguish whether the differences in mitochondrial metabolism were caused by the culture system or cell line differences?
- This is a very interesting comment and perhaps, it is a limitation of the present study, although it is easy to occur when comparing different cell lines. We wanted to use the optimal culture conditions established for each of the cell lines assuming that all of them will be in ideal conditions. We did not want to standardize the culture parameters because the effects caused by cell activity could be inaccurate and, above all, unrealistic.
There are already previous studies that evaluate metabolic parameters comparing different cell lines and they did the same as we did in this work, i.e., cultured each cell line under its best conditions of temperature, % CO2, and supplements (Lynch et al, 1987; De Angelis et al., 1999).
In summary, although it is true that there may be a limitation in the comparison, we believe that the results obtained are valid and lay the groundwork for other types of studies that do not necessarily have to be performed using at the same time the four cell lines studied in the present work. This fact has been added to the manuscript.
-Lynch, R.M.; Balaban, R.S. Energy Metabolism of Renal Cell Lines, A6 and MDCK: Regulation by Na-K-ATPase. Am. J. Physiol. 1987, 252, doi:10.1152/AJPCELL.1987.252.2.C225.
-De Angelis, I.; Rossi, L.; Pedersen, J.Z.; Vignoli, A.L.; Vincentini, O.; Hoogenboom, L.A.P.; Polman, T.H.G.; Stammati, A.; Zucco, F. Metabolism of Furazolidone: Alternative Pathways and Modes of Toxicity in Different Cell Lines. Xenobiotica. 1999, 29, 1157–1169, doi:10.1080/004982599238029.
- “Results & discussion”. The purpose of comparing inherently different cell lines is not clearly communicated to the reader. It is not enlightening to speak of difference as difference.
Improved as suggested
In addition, the use of four cell lines to test the differences in mitochondrial metabolism to guide peer researchers to select suitable cell lines for different experimental purposes (there are more than 900 fish cell lines) seems to assume that most peers do not know the different mitochondrial metabolic characteristics of different types of cell lines, which may be inconsistent with the reality.
The fact that a researcher assumes a fact does not mean that it cannot be measured, corroborated, and contrasted with precision by means of a type of analysis, as has been done in this case by performing the stress myth test. This work provides quantitative data using a relatively new technique rarely used in aquatic animals.
Round 2
Reviewer 2 Report
I understood the author's main ideas and realized the value of this research. However, personally, I still think this study is too general and may not give readers much direct reference value. At the same time, the experimental procedures and data statistics involved in this study are valuable to the peers and have their value as communication. I therefore accept the publication of this article in its present form.
