PONE-D-24-37080Respiratory pathology in the mdx/utrn-/- mice: a murine model for Duchenne Muscular Dystrophy (DMD)PLOS ONE

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Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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5. Review Comments to the Author

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Reviewer #1: In their work, M. Hernandez-Rodriguez et al. performed a systematic analysis of the respiratory abilities of mdx/utrn-/- mice, an animal model closely recapitulating the pathology of human patients suffering from Duchenne Muscular Dystrophy (DMD). Respiratory muscle weakness, ultimately leading to respiratory failure, is a serious and life-threatening aspect of this fatal X-linked neuromuscular disorder. Herein, the authors first evaluated life span and development of the pathology (weigth, kyphosis, grip strength), followed by measurements of respiratory function under normoxia (baseline) and after respiratory challenge with hypoxia and hypercapnia. Notably, respiratory muscle function and ventilation was significantly reduced in already in 6-weeks-old mdx/utrn-/- mice and was even more pronounced at end-stage. Finally, the work is complemented by ex vivo evaluation of the functional capacity of the diaphragm and histological analyses (H&E, Pico-Sirius red, immunostaining for dystrophin) of the major respiratory muscle, the diaphragm, as well as of tongue and tibialis anterior, and an evaluation of the neuromuscular junction. The experimental work is well-described and profoundly performed, the manuscript is well formulated. At some points, the authors should provide a more information on how and/or why they performed a certain experiment. Accordingly, there are several minor items that should be addressed:

1. Page 5, line 108: please define the exact age of mice at “end-stage” in the text.

2. Page 6, line 124: please provide a short explanation on why you choose these three muscles.

3. Page 6, lines 130-135: the numbering of Figures 6 and 7 is not correct; the Pico-Sirius red staining is discussed first (Figure 7 according to the figure legends), then the dystrophin immunostaining (Figure 6). Please re-arrange the figures according to the main text.

4. Page 6, lines 136-142: are the neuromuscular junctions in mdx/utrn-/- tongue and tibialis anterior fragmented as well? Why did the author just investigate the diaphragm here?

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Reviewer #1: No

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Respiratory pathology in the mdx/utrn-/- mouse: a murine model for Duchenne Muscular Dystrophy (DMD)

PONE-D-24-37080R1

Dear Dr. ElMallah,

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,

Gerhard Wiche, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments:

In addressing point 4 of reviewer 1 the authors may want to change the chapter title to "Analysis of diaphragmatic neuromuscular junctions" and streamline the text by combining sentence 1 and 2 (lines141-143).

Reviewers' comments: