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Biochem Biophys Res Commun ; 525(4): 1011-1017, 2020 05 14.
Article in English | MEDLINE | ID: mdl-32178872

ABSTRACT

In seminiferous epithelium, tight junctions (TJs) between adjacent Sertoli cells constitute the blood-testis barrier and must change synchronically for germ cells to translocate from the basal to the adluminal compartment during the spermatogenic cycle. Rho GTPase activation through stimulation with specific L-selectin ligands has been proposed to modulate tight junctional dynamics. However, little is known regarding the role of Ca+2 dynamics in Sertoli cell and how Ca+2 relays L-selectin signals to modulate Rho GTPase activity in Sertoli cells, thus prompting us to investigate the Ca+2 flux induced by L-selectin ligand in ASC-17D cells. Using fluorescent real-time image, we first demonstrated the increase of intracellular Ca+2 level following L-selectin ligand stimulation. This Ca+2 increase was inhibited in ASC-17D cells pretreated with nifedipine, the L-type voltage-operated Ca+2 channel (VOCC) blocker, but not mibefradil, the T-type VOCC blocker. We then demonstrated the up-regulation of Rho and Rac1 in ASC-17D cells following the administration of L-selectin ligand, and the pre-treatment with nifedipine, but not mibefradil, prior to L-selectin ligand-binding abolished the activation of both Rho and Rac1. Together, we conclude that the activation of L-selectin induces Ca+2 influx through the L-type VOCC, which up-regulates Rho and Rac1 proteins, in ASC-17D cells.


Subject(s)
Calcium/metabolism , L-Selectin/metabolism , Sertoli Cells/metabolism , Spermatozoa/metabolism , rho GTP-Binding Proteins/metabolism , Animals , Calcium Channel Blockers/pharmacology , Calcium Channels , Cell Line , Ligands , Male , Mibefradil/pharmacology , Nifedipine/pharmacology , Optical Imaging , Rats , Sertoli Cells/drug effects , Sertoli Cells/enzymology , Signal Transduction/drug effects , Signal Transduction/physiology , Spermatogenesis/drug effects , Spermatogenesis/genetics , rac1 GTP-Binding Protein/metabolism , rho GTP-Binding Proteins/genetics
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