Reply to Kissane and Eggington
2021; American Physical Society; Volume: 321; Issue: 6 Linguagem: Inglês
10.1152/ajpcell.00393.2021
ISSN1522-1563
AutoresNadav Tropp, Jennifer E. Gilda, Shenhav Cohen,
Tópico(s)Cerebral Palsy and Movement Disorders
ResumoLetter to the EditorReply to Kissane and EggingtonNadav Tropp, Jennifer E. Gilda, and Shenhav CohenNadav TroppFaculty of Biology, Technion Institute of Technology, Haifa, Israel, Jennifer E. GildaFaculty of Biology, Technion Institute of Technology, Haifa, Israel, and Shenhav CohenFaculty of Biology, Technion Institute of Technology, Haifa, IsraelPublished Online:07 Dec 2021https://doi.org/10.1152/ajpcell.00393.2021MoreSectionsPDF (191 KB)Download PDFDownload PDFPlus ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInEmail to the editor: We are grateful for the opportunity to further emphasize the novelty of our recently published methodology for measurement of cell size (1), and appreciate the comments on our paper by Kissane and Egintonn (2).Although a broader overview of the field could have been useful (3–17), a thorough and robust comparison between all published methodologies is simply outside the scope of a methodology paper and should be a subject for a dedicated review. The use of confocal microscopy is not a requirement of the Imaris software, which can process images taken by nonconfocal microscopes. Nor is the Imaris software being commercial necessarily a disadvantage, as it is an established standard in many microscopy centers around the world, and is thus already familiar and readily available for researchers.In Gilda et al. (1), we do not question the statistical validity of random sampling of fibers. We analyze whole muscle sections because the software is capable of processing them quickly and easily, providing data on the entire fiber population. Analysis of the entire muscle section is also an advantage in cases where random sampling would prove deleterious, such as when total fiber count is paramount (e.g., to validate hyperplasia). Imaris can also easily be used to analyze certain areas of a muscle section to obtain information on structural heterogeneities. This paper's avoidance of random sampling should not be conflated with the process of manually removing offending areas of a cross section, which must be removed for technical reasons, such as smeared edges or torn fibers, whose presence is a practical reality in most specimens.The use of a reporter protein (e.g., GFP) (18–20) in muscle transfection holds an important advantage over immunoreactivity by incorporating the fiber's transfection status into the semiautomated pipeline. The process allows quick and easy identification of transfected cells and quantitation of transfection efficiency, without relying on any changes to fibers' surface marker expression or the need for immunostaining.Kissane and Egintonn state that skewness is not a new statistic in the field, but do not cite publications reporting skewness values for fiber size, and we are likewise unaware of skewness being reported thus far. It is understandably difficult to provide literature examples for something we introduce as a novel, and discovering and validating conditions that lead to a shift in skewness is outside the scope of the paper. We offer a hypothetical example for when this statistic may prove useful: a change in size for only a fraction of the fibers that may not be reflected by a change in the mean or median but is biologically relevant. In a field where mean, median, and standard deviation are generally reported to describe distributions of hundreds to thousands of data points, we offer skewness and other statistical tests as means to report shifts in fiber size values and distributions. Kissane and Egintonn fail to acknowledge the potential usefulness of the statistical tests and summary statistics we describe, such as skewness, which we believe offer a great improvement over the current reporting of fiber size values.DISCLOSURESNo conflicts of interest, financial or otherwise, are declared by the authors.AUTHOR CONTRIBUTIONSN.T., J.E.G., and S.C. drafted manuscript; edited and revised manuscript; approved final version of manuscript.REFERENCES1. Gilda JE, Ko J-H, Elfassy A-Y, Tropp N, Parnis A, Ayalon B, Jhe W, Cohen S. A semi-automated measurement of muscle fiber size using the Imaris software. Am J Physiol Cell Physiol 321: C615–C631, 2021. doi:10.1152/ajpcell.00206.2021. Link | ISI | Google Scholar2. Kissane R, Eggingt S. Do we need another semi-automated approach to measure muscle fiber cross-sectional area? Am J Physiol Cell Physiol, 2021. doi:10.1152/ajpcell.00352.2021.Link | Google Scholar3. Li Y, Yang Z, Wang Y, Cao X, Xu X. 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USP1 deubiquitinates Akt to inhibit PI3K-Akt-FoxO signaling in muscle during prolonged starvation. EMBO Rep 21: e48791, 2020. doi:10.15252/embr.201948791.Crossref | PubMed | ISI | Google ScholarAUTHOR NOTESCorrespondence: S. Cohen ([email protected]ac.il). Previous Back to Top FiguresReferencesRelatedInformationRelated articlesDo we need another semiautomated approach to measure muscle fiber cross-sectional area? 07 Dec 2021American Journal of Physiology-Cell Physiology More from this issue > Volume 321Issue 6December 2021Pages C1084-C1085 Crossmark Copyright & PermissionsCopyright © 2021 the American Physiological Society.https://doi.org/10.1152/ajpcell.00393.2021PubMed34874767History Received 1 November 2021 Accepted 1 November 2021 Published online 7 December 2021 Published in print 1 December 2021 Keywordsautomated analysiscell sizecross-sectional areaImaris softwaremuscle atrophyPDF download Metrics Downloaded 282 times
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