STUDY ON THE FORMATION OF SPECIALIZED INTER-SERTOLI CELL JUNCTIONS IN VITRO

摘要: An in vitro culture system using Sertoli cells was employed to assess the expression of component genes pertinent occluding junctions (OJ) (such as zonula occludens-1, ZO-1), anchoring (AJ) N-cadherin and beta-catenin), communicating gap (GJ) connexin 33, Cx33) when they are being formed vitro. Freshly isolated from 20-day-old rats with a purity greater than 90% were cultured either at low- (2.5 x 10(4) cells/cm(2)) or high-cell density (0.6 10(6) on Matrigel-coated dishes for 7 days allow establishment specialized junctions. In low cell cultures, OJ such tight did not form during entire period assessed by transepithelial electrical resistance (TER). high there an increase ZO-1 1 3 preceding day 4. When both densities, transient expression, which peaked 4-5, suggesting time course AJ may overlap OJ. A significant beta-catenin also detected 5-7 but cultures. The Cx33 enhanced 4-5 These results suggest that OJ, AJ, GJ between whereas cannot be full-length cDNA clone coding rat testicular isolated. deduced amino acid sequence yielded 781 polypeptide displayed 99.9% identity mouse homolog. Conditioned medium germ induced dose-dependent stimulation affect N-cadherin/beta-catenin-mediated signal transduction pathway. summary, this study illustrates several target can used molecular markers monitor inter-Sertoli junction formation. This should applicable screen new male contraceptives targeted interference formation disrupting timely necessary and/or maintenance.