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Molecules
Volume 28
Issue 10
10.3390/molecules28104092
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Article
Peer-Review Record

Biocytin-Labeling in Whole-Cell Recording: Electrophysiological and Morphological Properties of Pyramidal Neurons in CYLD-Deficient Mice

Molecules 2023, 28(10), 4092; https://doi.org/10.3390/molecules28104092
by Shuyi Tan 1,2,†, Xiuping Mo 1,†, Huihui Qin 1,†, Binbin Dong 2, Jiankui Zhou 1, Cheng Long 2 and Li Yang 1,*
Reviewer 1:
Chandan Sona
Reviewer 2: Anonymous
Molecules 2023, 28(10), 4092; https://doi.org/10.3390/molecules28104092
Submission received: 30 March 2023 / Revised: 27 April 2023 / Accepted: 9 May 2023 / Published: 15 May 2023
(This article belongs to the Special Issue Functional Proteomics in Cell Biology and Beyond)

Round 1

Reviewer 1 Report

Shu-yi Tan et.al paper entitled " Biocytin-labeling in whole-cell recording: electrophysiological and morphological properties of pyramidal neurons in CYLD-deficient mice” is a methodological paper. I have few concerns regarding the paper.

Can the authors show that there were no morphological changes in neurons before and after completing the whole process of recording and labelling? I see that AAV virus expresses EGFP and those figures colocalizing with Biocytin is missing. I will like the authors to include that panel also.

Minor concerns:

1.     Please change on of the images as Fig1E and Fig 4D is same. Include other pictures.

2.     Screenshots of ImageJ (fig 5A-C) panel could be moved into the supplement and start fig5 with D. The detail of method is already there is the methods section no need to provide screenshot of the software.

3.     Fig 6F, WT mice images has less dendritic arborization than the KO mice. Can the authors change the image and select the suitable one for representation based on their analysis.

The quality of English can be improved and is average.

Author Response

Please see the attachment. Thank you.

Author Response File: Author Response.docx

Reviewer 2 Report

The first part, comprising of AAV-mediated GFP tagging, whole-cell patch, biocytin-streptavidin histology, and morphometric analyses, is fairly standard and practiced well by the authors. While this is OK, it is rather confusing to put MSN data since the second part of the manuscript is about PNs in CTKD-KO. The authors might consider putting the first part as a separate protocol manuscript. The second part of the paper reports that M1 L5 PNs in CYLD-KO is more excitable while the spine density is lower. Overall, this might mean that the input resistance of CYLD-KO PNs is higher than wild type. It is interesting that this phenotype is not observed in the BLA in a prior study (referenced as [33]). No mechanism is discussed.

Author Response

Please see the attachment. Thank you.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

All my comments were addressed. I have no further concerns.

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