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Developmental expression patterns of alpha1H T-type Ca2+ channels during spermatogenesis and organogenesis in mice
Weon-Young Son1, Ching-Tack Han, Jae-Ho Lee
1Department of Life Science, Sogang University, Seoul 121-742, Korea.
Abstract:
The objectives of the present study were to investigate the expression patterns of T-type Ca2+ channel mRNA during spermatogenesis and organogenesis in mice. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to identify the subtypes of calcium channels present in the round spermatids isolated from mouse testes by flow cytometry. Transcripts of L-type (alpha1D), non-L-type (alpha1E) and T-type Ca2+ channels were detected in round spermatids. Analysis of PCR products of T-type Ca2+ channels indicated that only alpha1H subunits were detected in round spermatids. The appearance and differential distribution of alpha1H T-type Ca2+ channel mRNA during mouse spermatogenesis and postimplantation embryogenesis (embryonic (E) days E9, E12, E15) were investigated by in situ hybridization with digoxigenin-labeled RNA probes coupled with alkaline phosphatase detection. In testes from adult and immature mice (postnatal 2 and 3 weeks), alpha1H T-type Ca2+ channel mRNA was expressed in all developing germ cells and sertoli cells. On E9 and E12, tissues of the central nervous system, such as the telencephalon, expressed alpha1H T-type Ca2+ channel mRNA. On E15, signals were detected throughout all organs of the embryo. These findings indicate that the expression of alpha1H T-type Ca2+ channels is spatio-temporally regulated during spermatogenesis and organogenesis.
Insights
The alpha1H T-type calcium channel mRNA is present in mouse germ cells and Sertoli cells during spermatogenesis. Its expression is also spatio-temporally regulated in the central nervous system and other organs during embryonic development.
Area of Science:
- Molecular Biology
- Developmental Biology
- Neuroscience
Background:
- T-type calcium channels play crucial roles in cellular excitability and proliferation.
- Understanding the expression patterns of specific calcium channel subunits is vital for comprehending developmental processes.
Purpose of the Study:
- To investigate the expression patterns of T-type calcium channel mRNA during mouse spermatogenesis and organogenesis.
- To identify specific T-type calcium channel subunits expressed in round spermatids.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) to detect calcium channel mRNA in round spermatids.
- Flow cytometry for isolating round spermatids from mouse testes.
- In situ hybridization to analyze the spatio-temporal distribution of alpha1H T-type calcium channel mRNA in developing embryos and testes.
Main Results:
- Transcripts for L-type (alpha1D), non-L-type (alpha1E), and T-type calcium channels were found in round spermatids.
- Only alpha1H T-type calcium channel subunits were detected in round spermatids.
- Alpha1H T-type calcium channel mRNA was expressed in germ cells and Sertoli cells in adult and immature mouse testes.
- During embryogenesis, alpha1H T-type calcium channel mRNA was detected in the central nervous system (telencephalon) on embryonic days 9 and 12, and throughout all embryonic organs by day 15.
Conclusions:
- The alpha1H T-type calcium channel is expressed in mouse round spermatids.
- The expression of alpha1H T-type calcium channel mRNA is spatially and temporally regulated during spermatogenesis.
- Alpha1H T-type calcium channel expression is also spatio-temporally regulated during mouse organogenesis, particularly in the developing central nervous system.