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Updated: Aug 14, 2026
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
CD2 is a surface marker for mouse and rat spermatogonial stem cells
Mito Kanatsu-Shinohara1,2, Guiying Chen1, Hiroko Morimoto1
1Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Abstract:
The spermatogonial stem cell (SSC) population in testis is small, and the lack of SSC markers has severely handicapped research on these cells. During our attempt to identify genes involved in SSC aging, we found that CD2 is expressed in cultured SSCs. Flow cytometric analysis and spermatogonial transplantation experiments showed that CD2 is expressed in SSCs from mature adult mouse testes. Cultured SSCs transfected with short hairpin RNAs (shRNAs) against CD2 proliferated poorly and showed an increased frequency of apoptosis. Moreover, functional analysis of transfected cells revealed impairment of SSC activity. Fluorescence activated cell sorting and spermatogonial transplantation experiments showed that CD2 is expressed not only in mouse but also in rat SSCs. The results indicate that CD2 is a novel SSC surface marker conserved between mouse and rat SSCs.
Insights
CD2 is identified as a novel marker for spermatogonial stem cells (SSCs) in mice and rats. This discovery aids in understanding SSC aging and function, overcoming previous research limitations.
Area of Science:
- Reproductive biology
- Stem cell research
- Molecular biology
Background:
- Spermatogonial stem cells (SSCs) are crucial for sperm production but are difficult to study due to their small numbers and lack of specific markers.
- Research into SSC aging and function has been significantly hindered by these limitations.
Purpose of the Study:
- To identify novel surface markers for spermatogonial stem cells (SSCs).
- To investigate the role of CD2 in SSC aging and function.
Main Methods:
- Flow cytometry and fluorescence-activated cell sorting (FACS) were used to detect CD2 expression.
- Spermatogonial transplantation assays were performed to assess SSC function.
- Short hairpin RNA (shRNA) technology was employed to knock down CD2 expression in cultured SSCs.
Main Results:
- CD2 was found to be expressed on SSCs in mature adult mouse testes.
- Knockdown of CD2 in cultured SSCs resulted in reduced proliferation, increased apoptosis, and impaired SSC activity.
- CD2 expression was also observed on rat SSCs, indicating conservation across species.
Conclusions:
- CD2 is a novel and conserved surface marker for spermatogonial stem cells (SSCs) in both mice and rats.
- CD2 plays a significant role in maintaining SSC proliferation, survival, and function.
- The identification of CD2 as an SSC marker opens new avenues for studying SSC biology and aging.
