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《Acta Zoologica Sinica》 2003-04
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Age-specific changes in androgen receptors in rat testis*

WANG Xiao-Yun ZHANG Jian LI Jian DUAN Xiang-Lin ** (College of Life Science,Hebei Normal University,Shijiazhuang 050016,China)  
In order to study age-specific changes in androgen receptors (ARs) we investigated changes in the expression of AR's in 60 SD rats, ranging in age from neonatal to 25 months. There were ten age groups: neonatal, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 12 months and 25 months. After rats were killed, their testes were immediately removed and fixed in Bouin's solution for up to 24 hours. Testes were dehydrated by a series of ethanol rinses, cleared in xylene and embeded in paraffin after which they were cut into 6 μm sections on a microtome. Immunocytochemistry analyses were performed after deparaffinage and rehydration. Primary antibodies were diluted 1∶100 in 0 01 mol/L PBS (pH 7 4). The control sections had the primary antibody omitted. Before incubation with the primary antibody, sections were pretreated in goat serum for 1 hour at 37℃. Following incubation with the primary antibody overnight at 4℃, sections were treated for 30 minutes in a biotinylated secondary antibody that matched the species of the primary antibody, followed by streptavidin/peroxidase complex incubation for 30 minutes at 37℃. Between incubation steps, the sections were rinsed three times for 15 minutes in PBS, but there was no PBS rinse between serum incubation and primary antibody incubation. Bound peroxidase was revealed by incubating the sections in 3, 3'-diaminobenzidine for 3-5 minutes. Sections were rinsed in PBS to stop the reaction, then dehydrated in a series of increasing concentrations of ethylalcohol. After being cleared in xylene, sections were coverslipped with a permanent mounting medium. We determined the average optical density of AR positive cells with an Image Analysis System. All data were analyzed using Excel and statistical significance determined with ANOVA. There were many kinds of AR-immunoreactive cells in normal rat testis, such as Leydig cells,endothelial cells,myoid cells,Sertoli cells, as well as spermatogenic cells. The distribution and staining intensity of AR-immunoreactive cells changed greatly with age. Our results show that: (1) There was a weakly positive expression of AR in Leydig cells in the first 3 weeks after birth. Positive expression of AR in Leydig cells peaked after 1 month but decreased markedly after 2 months after which it then increased gradually. (2) A weakly positive expression of AR in myoid cells was observed from in the first 2 weeks after birth. The positive expression of AR increased prominently in 3 week old rats but had decreased in 3 month old individuals,reaching its lowest level in 25 month old rats. (3) A strongly positive expression of AR in endothelial cells was observed in rats between 3 weeks and 2 months of age. It decreased prominently in 3 month old rats but there were no significant changes between 3 months and 25 months of age. (4) The positive expression of AR in Sertoli cells appeared at 1 month of age. In mature rats, the positive expression of AR in Sertoli cells changed with the spermatogenic cycle, but there was no expression of AR in Sertoli cells at 25 months. (5) The positive expression of AR in prospermatogonium cells appeared in neonatal rats. It appeared in spermatoons after 2 weeks, was observed in spermatogonia after 1 month and appeared in sperm after 2 months. But there was no expression of AR in spermatogenic cells at 25 months .
【Fund】: 河北省自然科学基金资助项目 (No. 3 0 0 164 )~~
【CateGory Index】: Q492
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