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TGFβ superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
Catherine Itman1, Chin Wong, Penny Af Whiley
1Department of Biochemistry and Molecular Biology; School of Biomedical Sciences; Monash University; Melbourne, Australia.
Abstract:
Transforming growth factor-beta (TGFβ) superfamily ligands are produced by and act upon testicular cells to control testis morphogenesis and adult fertility. Ligand production changes during testis development and dysregulated signaling affects the number of cells comprising each lineage and their development, with several components of this diverse signaling pathway linked to male infertility. To test the hypothesis that TGFβ superfamily signaling regulators are differentially expressed during mouse testis development, we surveyed expression of Hgs, Zfyve9, Smurf1 and Net25 by northern blot and in situ hybridization and SMURF2 and MAN1 by western blot and immunohistochemistry. Expression of these genes is highly regulated and differs between the first spermatogenic wave and adult spermatogenesis. Zfyve9 transcripts were first detected in Sertoli cells and spermatogonia at 5 days post partum (dpp) whereas Hgs mRNA was first detected in pachytene spermatocytes at 15 dpp. Smurf1 mRNA was broadly expressed at 0 and 5 dpp but restricted to spermatogonia and early spermatocytes at 15 dpp and spermatogonia, spermatocytes and round spermatids in adults. SMURF2 was limited to gonocyte nuclei at birth but was nuclear in all cells at 5 dpp. SMURF2 was absent from 15 dpp differentiating spermatogonia and early spermatocytes but readily detected in adult pachytene spermatocytes and round spermatids. MAN1 and Net25 also had different expression profiles, with MAN1 undetectable at 5 dpp. Differential synthesis of signaling modulators explains how Sertoli cells and spermatogenic cells, which all possess TGFβ superfamily signaling machinery and reside within the same microenvironment, respond differently to the same ligand.
Insights
Regulators of transforming growth factor-beta (TGFβ) superfamily signaling show distinct expression patterns during mouse testis development. These changing levels of TGFβ regulators are crucial for controlling cell development and ensuring male fertility.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Molecular Endocrinology
Background:
- Transforming growth factor-beta (TGFβ) superfamily signaling is vital for testicular development and male fertility.
- Dysregulation of TGFβ signaling components is linked to male infertility.
- Understanding the temporal and spatial expression of TGFβ regulators is key to deciphering their roles in testis development.
Purpose of the Study:
- To investigate the hypothesis that TGFβ superfamily signaling regulators are differentially expressed during mouse testis development.
- To map the expression profiles of key regulators including Hgs, Zfyve9, Smurf1, Net25, SMURF2, and MAN1.
Main Methods:
- Northern blot and in situ hybridization were used to analyze mRNA expression of Hgs, Zfyve9, Smurf1, and Net25.
- Western blot and immunohistochemistry were employed to assess protein levels of SMURF2 and MAN1.
- Expression patterns were examined at various developmental stages: birth (0 days post partum - dpp), 5 dpp, 15 dpp, and adulthood.
Main Results:
- Expression of TGFβ regulators is highly regulated and changes significantly between the first spermatogenic wave and adult spermatogenesis.
- Specific genes like Zfyve9, Hgs, Smurf1, SMURF2, and MAN1 exhibited distinct temporal and cell-type-specific expression patterns.
- For example, Zfyve9 was detected in Sertoli cells and spermatogonia at 5 dpp, while Hgs appeared in pachytene spermatocytes at 15 dpp.
Conclusions:
- The differential expression of TGFβ signaling modulators during testis development provides a mechanism for cell-specific responses to TGFβ ligands.
- These precisely regulated expression patterns are essential for proper control of testis morphogenesis and the maintenance of adult fertility.
- Understanding these dynamic changes in signaling components offers insights into potential causes of male infertility related to TGFβ pathway disruption.
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