Course of c-myc mRNA expression in the regenerating mouse testis determined by competitive reverse transcriptase polymerase chain reaction.
The c-myc proto-oncogene is a reliable marker of the "G0-early G1" transition, and its down-regulation is believed to be necessary to obtain cellular differentiation. In murine spermatogenesis, the level of c-myc transcripts does not correlate with the rate of cellular division. Proliferation of supposed staminal spermatogonia to reproduce themselves is induced with a local 5 Gy X-ray dose in 90-day-old C57Bl/6 mice. c-myc quantification by a newly developed competitive reverse transcriptase polymerase chain reaction (RT-PCR) was carried out to follow the expression course of this proto-oncogene. Damage and restoration of spermatogenesis were analyzed at days 3, 6, 9, 10, 13, 30, and 60 after injury by relative testes/body weight determination and histological examination. Proliferative status was determined by histone H3 Northern blot analysis. c-myc mRNA level was 10 times higher after 3 days in the irradiated animals compared to the controls. An increasing number of copies were noted up to 10 days, but promptly decreased to the base level found for irradiated mice from 13 to 60 days. Interestingly, the expression of histone H3 detected S phase only in testes at 60 days from damage.
Amendola
R
R
Department of Environmental Sciences, Enea CR-Casaccia, Roma, Italy.
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001835:Body Weight; D017931:DNA Primers; D005091:Exons; D015870:Gene Expression / Q000528:radiation effects; D016259:Genes, myc; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009929:Organ Size; D016133:Polymerase Chain Reaction / Q000379:methods*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis*; D011519:Proto-Oncogenes; D018075:RNA, Complementary / Q000138:chemical synthesis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase; D012016:Reference Values; D012038:Regeneration; D012044:Regression Analysis; D013087:Spermatids / Q000166:cytology / Q000502:physiology / Q000528:radiation effects; D013090:Spermatocytes / Q000166:cytology / Q000502:physiology / Q000528:radiation effects; D013091:Spermatogenesis / Q000528:radiation effects; D013093:Spermatogonia / Q000166:cytology / Q000502:physiology / Q000528:radiation effects; D013737:Testis / Q000378:metabolism*; D013997:Time Factors; D014965:X-Rays
D017931:DNA Primers; D016271:Proto-Oncogene Proteins c-myc; D018075:RNA, Complementary; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase