Protein Arginine Methyltransferase 5 Is Necessary for Embryonic Development in Medaka Oryzias latipes
, Xueping Zhong 1 and Haobin Zhao 1,*
Round 1
Reviewer 1 Report
The manuscript entitled, “Prmt5 is necessary for embryonic development in medaka Oryzias latipes” deals about the role of prmt5 during embryogenesis via maintaining stemness and repressing apoptosis in medaka fish. Though the manuscript is well written and the experimental approach seems interesting, the lack of substantial evidence makes the manuscript weak. In addition, the following comments should be addressed by the authors.
1. In addition to quantification of the transcript levels of genes related to ES cells, PGCs, and apoptosis or cell death, is it essential to check mitochondrial function assay/ cytotoxic analysis: cell viability assay/ membrane permeability: apoptosis assay through flow cytometry/ brdU DNA proliferation test/ reactive oxygen species generation in vitro to confirm apoptosis.
2. It is also essential to analyze the protein level changes of a few related proteins to further add substantial value to the manuscript.
The authors need to address the above critical comments, if possible by adding experimental data. The manuscript certainly requires extensive revision failing which the manuscript is not suitable and hence, my decision is major revision.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The study was interesting and valuable. However, some sections should be revised.
The title shouldn’t contain abbreviations.
All abbreviations should be defined as first mentioned.
Please provide the Latin name of each fish species first mentioned.
Methods:
More information should be provided about the duration and time of the experiment and the total number of male and female fish used. Source of these fish (from where were they obtained?) Management and rearing of fish during the experiment. Dealing with eggs and embryos. Where were the fish reared, Ponds or aquaria? How were the eggs collected? The number of eggs spawned during the experiment. Age of maturity on which fish on each cycle were mated? How samples from eggs and embryos were collected.
Table 2: rewrite the p-value to a decimal number.
Results and discussion: the genes related to stemness and 253 apoptosis were down- or upregulated: please clarify when down and when up?
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
Review - fishes-2053815
The manuscript titled: „Prmt5 is necessary for embryonic development in medaka Oryzias latipes” is very interesting and valuable. I have suggested minor revisions based on my suggestions included below. In my opinion, this paper is acceptable for publication after revision.
Materials and methods section - Please provide:
1. The project identification code of the ethics committee
that has approved your research.
2. Information about fish spawners' length weight age, moreover the number of fish spawners and their origin. The number of fish from each generation.
3. procedure of microinjection with characteristics of applied equipment.
Line 181 – the name of the molecular marker and producer should be included
Line 199-201 – the information about number of mutant offspring should be included
Line 205 – the number of F1 embryos in brackets should be added
Figure 6 – Please indicate genes upregulated and downregulated using arrows
I was wondering if the ploidy level was studied? Please provide such information.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
No further comments. Thank you
