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Isolation of Sertoli Cells and Peritubular Cells from Rat Testes
Published on: February 8, 2016
Dynamic testicular adhesion junctions are immunologically unique. I. Localization of p120 catenin in rat testis
Kamin J Johnson1, Kim Boekelheide
1Department of Pathology and Laboratory Medicine, Brown University, 175 Meeting Street, Providence, Rhode Island 02912, USA. kamin_johnson@brown.edu
Insights
p120 catenin is a key component of desmosome-like junctions in the rat testis, with specific isoforms localizing to distinct cell junctions. This study reveals unique molecular compositions in Sertoli-germ cell junctions, crucial for understanding testis development and function.
Area of Science:
- Reproductive Biology
- Cell Biology
- Molecular Biology
Background:
- The seminiferous epithelium relies on diverse junctions for cell adhesion.
- The molecular composition of Sertoli-Sertoli and Sertoli-germ cell junctions is not fully understood.
- Catenins are critical for regulating adhesion strength at adherens junctions.
Purpose of the Study:
- To investigate the localization and role of p120 catenin (p120) in rat testis junctions.
- To determine the molecular composition of inter-Sertoli and Sertoli-germ cell junctions.
- To identify specific p120 isoforms associated with different junction types.
Main Methods:
- Immunofluorescence using monoclonal antibodies (5A7, 8D11, 15D2) on rat testis cryosections and touch preparations.
- Immunoprecipitation of testis homogenates to identify p120 isoforms.
- Colocalization studies with beta-catenin and plectin.
Main Results:
- At least four p120 catenin isoforms are expressed from postnatal day 7 through adulthood.
- p120 catenin is present in both inter-Sertoli and Sertoli-germ cell junctions.
- Antibodies 5A7 and 8D11 colocalize with beta-catenin and plectin at inter-Sertoli and Sertoli-spermatocyte junctions, suggesting p120 is part of desmosome-like junctions.
- Antibody 15D2 specifically labels Sertoli-round spermatid and newly formed inter-Sertoli junctions in a dynamic, stage-specific pattern.
Conclusions:
- p120 catenin is a component of most desmosome-like junctions in the rat testis.
- Different cell types exhibit unique molecular compositions within their desmosome-like junctions.
- The dynamic localization of p120 suggests its involvement in the remodeling of these junctions during spermatid differentiation.
Abstract:
In the seminiferous epithelium, morphologically diverse junctions mediate inter-Sertoli and Sertoli-germ cell adhesive contact, but the molecular composition of such junctions is not well known. At prototypical adherens junctions, proteins termed catenins bind to the intracellular domain of classic cadherins and regulate the strength of adhesion. Using a panel of monoclonal antibodies (5A7, 8D11, and 15D2), p120 catenin (p120) was localized in postnatal and adult rat testis cryosections and touch preparations by immunofluorescence. Immunoprecipitation of testis homogenates showed that at least four p120 isoforms were expressed from Postnatal Day 7 through adulthood. Both inter-Sertoli and Sertoli-germ cell junctions were p120-positive, however, individual p120 monoclonals were localized to specific junctions. The 5A7 and 8D11 antibodies colocalized with beta-catenin and plectin at inter-Sertoli and Sertoli-spermatocyte junctions. At inter-Sertoli junctions, p120 was juxtaposed to but did not colocalize with f-actin. Thus, p120 is likely a component of inter-Sertoli desmosome-like junctions. In contrast, the 15D2 monoclonal antibody specifically immunostained Sertoli-round spermatid and inter-Sertoli cell junctions in a dynamic pattern. From the time that round spermatids form to their differentiation into elongate spermatids, Sertoli-round spermatid 15D2 immunostaining cycled from a single mass to a curvilinear pattern, and finally to punctate structures scattered throughout the epithelium. This localization and stage-specific immunostaining pattern indicated that 15D2 recognized Sertoli-round spermatid desmosome-like junctions. Between Sertoli cells, 15D2 immunostained newly formed junctions (at Postnatal Days 21 through 43), but not mature junctions in the adult. From these data, we conclude that p120 is a component of most, if not all, desmosome-like junctions, and that desmosome-like junctions between different cell types contain a unique molecular composition.
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