PLZF^posc-KIT^pos-delineated A1-A4-differentiating spermatogonia by subset and stage detection upon Bouin fixation  

作　　者：Rui-Ling Tang Li-Qing Fan 

机构地区：[1]Institute of Reproduction and Stem Cell Engineering, School of Basic Medicine Science, Central South University, Changsha 410078, China

出　　处：《Asian Journal of Andrology》2019年第3期309-318,共10页亚洲男性学杂志（英文版）

基　　金：the grants from the National Key Research and Development Program of China (Project No. 2016YFC1000200);the National Natural Science Foundation of China (Project No. 31472054).

摘　　要：While hallmarks of rodent spermatogonia stem cell biomarkers' heterogeneity have recently been identified, their stage and subset distributions remain unclear. Furthermore, it is currently difficult to accurately identify subset-specific SSC marker distributions due to the poor nuclear morphological characteristics associated with fixation in 4% paraformaldehyde. In the present study, testicular cross-sections and whole-mount samples were Bouin fixed to optimize nuclear resolution and visualized by immunohistochemistry (IHC) and immunofluorescence (IF). The results identified an expression pattern of PLZFhighc-KITpos in A1 spermatogonia, while A2–A4-differentiating spermatogonia were PLZFlowc-KITpos. Additionally, this procedure was used to examine asymmetrically expressing GFRA1 and PLZF clones, asymmetric Apr and false clones were distinguished based on the presence or absence of TEX14, a molecular maker of intercellular bridges, despite having identical nuclear morphology and intercellular distances that were <25 μm. In conclusion, this optimized Bouin fixation procedure facilitates the accurate identification of spermatogonium subsets based on their molecular profiles and is capable of distinguishing asymmetric and false clones. Therefore, the findings presented herein will facilitate further morphological and functional analysis studies and provide further insight into spermatogonium subtypes.

关 键 词：asymmetric division cellular homolog of feline sarcoma viral oncogene v-kit false clones GLIAL CELL line-derived NEUROTROPHIC factor receptor alpha 1 PROMYELOCYTIC leukemia zinc finger SPERMATOGONIA stem CELL 

分 类 号：R[医药卫生]