Possible involvement of MLH3, MLH1 & MSH4 genes in prophase I progression in human spermatogenesis [Possível envolvimento dos genes MLH3, MLH1 e MSH4 na Progressão da Profase I na Esperatogénese Humana]

• Type: Article in International Scientific Journal
• Date: 2005

Authors

Ferras, C (Author) Other; Fernandes, S (Author) Other; sousa, m (Author) ICBAS; barros, a (Author) FMUP

Journal

Title: Revista Iberoamericana de Fertilidad y Reproduccion Humana

Vol. 22

Pages: 67-74

ISSN: 1132-0249

Publisher: Sociedad Espanola de Fertilidad

Indexing

Scopus - 0 Citations

Other information

Authenticus ID: P-007-D9X

Abstract (EN): a) Objectives: Identification and quantitative analysis of genic expression in the different cell stages of the human seminiferous epithelium. b) Study design: Quantitative mRNA analysis of MLH3, MLH1 and MSH4 in isolated human stage-specific male germ cell populations. c) Study localization: University of Porto, Portugal. d) Characteristics of the study population: Under informed consent, cells were isolated from treatment testicle biopsies of two men with psychologic anejaculation, normal karyotypes, absence of Y chromosome microdeletions and conserved spermatogenesis. e) Main interventions: Sertoli cells, A spermatogonia, primary spermatocytes, secondary spermatocytes and round spermatids were isolated by micromanipulation. mRNAs were isolated with Rneasy Mini Kit and converted to cDNA with SuperScript II and random primers. The levels of expression were determined by real-time PCR with SybrGreen detection and GAPDH/-Actin as normalizer genes. f) Main outcomes: The pattern curves for each expressed gene were obtained with serial RNA dilutions from a sample of whole testicular tissue from each patient. The level of expression of each gene in each cell population was then determined from the respective pattern curve. The ratio between the expression levels of the target and normalizer genes gives the relative expression of each studied gene. g) Results: Gene expression was not detected in Sertoli cells, secondary spermatocytes and round spermatids. Gene expression was maximally detected in primary spermatocytes, with MLH3 showing the highest expression level. h) Conclusions: Data suggest that the DNA repair system is active during human male meiosis I.

Language: Spanish

Type (Professor's evaluation): Scientific