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KMID : 0356420000180010033
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Journal of Korean Andrology
2000 Volume.18 No. 1 p.33 ~ p.40
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Expression of Testis Specific Genes of Male Germ Cells in Pre-pubertal Mouse using in vitro Culture System
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Lee Ho-Joon
Kim Myo-Kyung
Ko Duck-Sung
Kang Hee-Gyoo
Kim Dong-Hoon
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Abstract
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Purpose: To establish cultures of premeiotic mouse spermatogenic cells that could allow study of the meiotic process in vitro and to investigate expression of sperm-specific genes in premeiotic and postmeiotic division.
Material and Methods: Male ICR mice were used. To obtain the premeiotic spermatogenic cells, we examined the histologic appearance of the germ cells in hematoxylin-eosin-stained testis sections during prepubertal development. To confirm the identity of the premeiotic cells, we performed reverse transcriptase-polymerase chain
reaction(TR-PCR) for the genes for LDH-C4, acrosin, protamine-2(P-2), and SP-10, which are testis-specific marker proteins. In order to establish the culture system, the premeiotic spermatogenic cells were cocultured with mouse Sertoli cells in DMEM containing 10% fetal bovine serum (FBS), amino acids, FSH, and testosterone at 32 for 6 days. After 2 days of culture, RT-PCR was done to detect the sperm-specific genes.
Results: The four genes was not expressed in 10-day-old mice process, and round spematids were seen in the testis. Premeiotic germ cells isolated from 15-day-old mice expressed LDH-C4 and acrosin but not P-2 and SP-10, which are post-meiotic marker proteins. Beginning after 2 days of culture, expression of the p-2 and SP-10 genes was
detected in cultured premeiotic germ cells as in 15-day-old mice.
Conclusions: P-2 and SP-10 may be marker genes for the premeiotic stage of spermatogenesis. Premeiotic male germ cells are able to differentiate into postmeiotic forms during coculture with Sertoli cells.
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KEYWORD
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Meiosis, Mouse, Spematogenesis, Testis
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