Role of DNase Activity in Human Sperm DNA Fragmentation

Round 1

Reviewer 1 Report

The paper titled "Role of DNase Activity in Human Sperm DNA Fragmentation" explores the significance of DNase activity in sperm DNA fragmentation (SDF) and its implications for male fertility. This work provides a comprehensive overview of the factors contributing to SDF, emphasizing the multifactorial nature of the pathology and the potential role of enzymatic activity, particularly DNase, in DNA degradation. The conclusion underscores the importance of considering enzymatic factors in understanding SDF and its impact on reproductive outcomes. Overall, the paper offers valuable insights into the mechanisms underlying male infertility and highlights the need for further research into the role of DNase activity in sperm DNA integrity.

While the paper provides a thorough discussion of the role of DNase activity in sperm DNA fragmentation, further clarification on sequence-dependent (e.g. DNA Crookedness ) features would broaden the interest of the findings by linking it with other gene expression endeavours.

Overall, this work is a valuable contribution to the readership of biomolecules, and thus I recommend publication.

Author Response

**Dear Reviewer 1, Thank you for comments and suggestions. Please find our responses to your comments below.

The paper titled "Role of DNase Activity in Human Sperm DNA Fragmentation" explores the significance of DNase activity in sperm DNA fragmentation (SDF) and its implications for male fertility. This work provides a comprehensive overview of the factors contributing to SDF, emphasizing the multifactorial nature of the pathology and the potential role of enzymatic activity, particularly DNase, in DNA degradation. The conclusion underscores the importance of considering enzymatic factors in understanding SDF and its impact on reproductive outcomes. Overall, the paper offers valuable insights into the mechanisms underlying male infertility and highlights the need for further research into the role of DNase activity in sperm DNA integrity.

**Thank you for kind appraisal.

While the paper provides a thorough discussion of the role of DNase activity in sperm DNA fragmentation, further clarification on sequence-dependent (e.g. DNA Crookedness ) features would broaden the interest of the findings by linking it with other gene expression endeavours.

**A new paragraph has been added in which reference is made to the effect of seminal plasma DNases on DNA compacted in chromatin in comparison with the effect of treatment with recombinant DNase I.

Overall, this work is a valuable contribution to the readership of biomolecules, and thus I recommend publication.

** Thank you

Reviewer 2 Report

The manuscript is quite well written, and the approach and the results are given understandably and transparently.

There are some small issues:

Line 178, Why did you use "Reference 78" rather of writing the reference in manuscript format? If you are concerned about self-citation, it may have been clarified in the text.
You should write it in the text and utilize reference bibliography order for this citation.


The Abstract and Introduction place emphasis on "iatrogenic damage" to sperm. When reading, it appears that the study will discuss, test, or correlate with DNAse activity, but it never returns to the rest of the paper. I feel this is an essential consideration, but it is not relevant to this manuscript.

Figure 1 Legend, Line 199, PS should be SP (seminal plasma).

The manuscript examines the effect of DNAse on genomic DNA. To validate these data, a DNAse enzyme activity assay experiment should be performed separately.

Author Response

**Dear Reviewer 2, Thank you for comments and suggestions. Please find our responses to your comments below.

The manuscript is quite well written, and the approach and the results are given understandably and transparently.

**Thank you for kind appraisal.

There are some small issues:

Line 178, Why did you use "Reference 78" rather of writing the reference in manuscript format? If you are concerned about self-citation, it may have been clarified in the text.
You should write it in the text and utilize reference bibliography order for this citation.

**Thank you for bringing this to our attention. We have made the correction in manuscript and spotted another reference in the same figure legend that also need converting to the correct referencing style.

The Abstract and Introduction place emphasis on "iatrogenic damage" to sperm. When reading, it appears that the study will discuss, test, or correlate with DNAse activity, but it never returns to the rest of the paper. I feel this is an essential consideration, but it is not relevant to this manuscript.

**We have removed reference to iatrogenic damage and replaced it with the more general concept of post-ejaculation sperm manipulation. As the reviewer indicates the topic while important is not relevant to the current review.

Figure 1 Legend, Line 199, PS should be SP (seminal plasma).

**Corrected.

The manuscript examines the effect of DNAse on genomic DNA. To validate these data, a DNAse enzyme activity assay experiment should be performed separately.

**While we are sympathetic to what the reviewer is suggesting here, what is being requesting is beyond the scope of a “review” and would represent a significant work in its own right.