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Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse
Spermatogenesis generates heterologous cell populations which, if not distinguished clearly, often hinder mechanistic and etiological studies. Here, we present a protocol to identify and isolate populations of mouse spermatogenic cells, including spermatogonial stem cells (SSCs), spermatocytes, and...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9117932/ https://www.ncbi.nlm.nih.gov/pubmed/35600921 http://dx.doi.org/10.1016/j.xpro.2022.101398 |
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author | Zou, Qianxing Yang, Lele Qi, Huayu |
author_facet | Zou, Qianxing Yang, Lele Qi, Huayu |
author_sort | Zou, Qianxing |
collection | PubMed |
description | Spermatogenesis generates heterologous cell populations which, if not distinguished clearly, often hinder mechanistic and etiological studies. Here, we present a protocol to identify and isolate populations of mouse spermatogenic cells, including spermatogonial stem cells (SSCs), spermatocytes, and haploid spermatids. We also describe absolute quantification of mRNA copy numbers in SSCs. The isolated cells can be used for analyzing nascent protein synthesis and protein degradation, two main events that maintain cellular proteostasis important for healthy and long-term production of male gametes. For complete details on the use and execution of this protocol, please refer to Zou et al. (2021). |
format | Online Article Text |
id | pubmed-9117932 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-91179322022-05-20 Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse Zou, Qianxing Yang, Lele Qi, Huayu STAR Protoc Protocol Spermatogenesis generates heterologous cell populations which, if not distinguished clearly, often hinder mechanistic and etiological studies. Here, we present a protocol to identify and isolate populations of mouse spermatogenic cells, including spermatogonial stem cells (SSCs), spermatocytes, and haploid spermatids. We also describe absolute quantification of mRNA copy numbers in SSCs. The isolated cells can be used for analyzing nascent protein synthesis and protein degradation, two main events that maintain cellular proteostasis important for healthy and long-term production of male gametes. For complete details on the use and execution of this protocol, please refer to Zou et al. (2021). Elsevier 2022-05-14 /pmc/articles/PMC9117932/ /pubmed/35600921 http://dx.doi.org/10.1016/j.xpro.2022.101398 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Zou, Qianxing Yang, Lele Qi, Huayu Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title | Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title_full | Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title_fullStr | Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title_full_unstemmed | Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title_short | Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
title_sort | protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9117932/ https://www.ncbi.nlm.nih.gov/pubmed/35600921 http://dx.doi.org/10.1016/j.xpro.2022.101398 |
work_keys_str_mv | AT zouqianxing protocolforisolationandproteostaticanalysisofsubpopulationsofspermatogeniccellsinmouse AT yanglele protocolforisolationandproteostaticanalysisofsubpopulationsofspermatogeniccellsinmouse AT qihuayu protocolforisolationandproteostaticanalysisofsubpopulationsofspermatogeniccellsinmouse |