PONE-D-20-16873

How scars shape the neural landscape: key molecular mediators of TGF-β1’s anti-neuritogenic effects

PLOS ONE

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Antal Nógrádi, M.D., Ph.D., D.Sc.

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The paper is technically complex and hence challenging to resent.

I found the message was lost in the details.

It would be very helpful for the reader to have a "big picture" diagram as Figure 1.

Also, the author states that the magnification was 10x or 20X. It is more likely that a 10X or 20X objective was used and that is in combination with 10X binoculars so the magnification would be 100X and 200X. Please check this.

Reviewer #2: This study aimed to investigate the effect of TGF-β1 on intracellular signals associated with neurite outgrowth in the neuronal hybridoma cell line ND7/23. ND7/23 cell differentiated by addition of nerve growth factor. Cells extend neurites and express specific markers of sensory neurons. The authors used different methods to evaluate the impact of TGF-β1 on neurite outgrowth. TGF-β1 treatment inhibited neurite extension in differentiated ND7/23 cells and increased the phosphorylation of the collapsin response mediator protein-2 (CRMP2). Furthermore, both glycogen synthase kinase (GSK)-3β and Smad3 modulate phosphorylation of CRMP2 after treatment with TGF-β1.

The study provides evidence that TGF-β1 mediated inhibition of neurite outgrowth in sensory neurons appears to be regulated through a highly-conserved signaling pathway.

There are some minor points to be corrected:

1) Figure 1: Scale bar is missing in Figure 1A-C. Please insert one. Magnification indicators are not informative for the reader, please insert scale bars throughout the MS.

2) The authors write in the results’ section (Impact of TGF-β 1 on neurite outgrowth) “these cells were cultured with 50ng/ml rNGF for 1 day, the proportion of cells in Stage 1 decreased (two-tailed paired t-test for 0.5 versus 50 ng/ml rNGF conditions: t3=4.33, p=0.023), while the proportion of Stage 3 cells nearly doubled (two-tailed paired t-test: t3=3.58, p=0.037) – see pictures in Fig. 2E”. Figure 2E does not represent this statement. Please clarify.

3) Please indicate the significant difference in Figure 2D

4) What drug/treatment can block the effect of TGF-β1 in order to promote neurite outgrowth? Please discuss in detail. with specific details on molecular mechanisms.

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Reviewer #1: No

Reviewer #2: No

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How scars shape the neural landscape: key molecular mediators of TGF-β1’s anti-neuritogenic effects

PONE-D-20-16873R1

Dear Dr. Huxlin,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Antal Nógrádi, M.D., Ph.D., D.Sc.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: