Producción CyT
Biocell - THE PROTEIN KINASE D FAMILY REGULATE SPERMATOZOA CAPACITATION
Congreso
Autoría:
MARTÍNEZ LEÓN, EDUARDO ANTONIO ; Osycka-Salut, Claudia Elena ; Marin-Briggiler, CL ; Jabloñski, M. ; Buffone, Mariano ; Rey, OsvaldoFecha:
2023Editorial y Lugar de Edición:
INST HISTOL EMBRIOL-CONICETISSN:
0327-9545Resumen *
The protein kinase D family(PKDs) are serine/threonine kinases named PKD1, PKD2 and PKD3. These isoenzymes are involved in fundamental biological process regulation, including signal transduction, cytoskeleton remodeling, oxidative stress, among others. Sperm are specialized and transcriptionally inactive cells dependent on post-translational modifications, such as protein phosphorylation, to carry out their functions. These changes occur during the process called sperm capacitation. The presence and functions of PKDs in the male gamete have not been described. Our aim was to characterize the presence and function(s) of PKDs in these cells.Through indirect immunofluorescence(IIF) and western blot (WB), we observed that PKD1/2 is present and active(p-PKD) in bovine, equine, mouse, and human sperm in not capacitating (NC) and capacitating conditions (C). The PKD´s localization was similar in all the species studied: head and flagellum. Interestingly, p-PKD distribution pattern varies between species. In humans, it is in the middle piece and, in mouse it is in the main piece and head. Regarding PKD3, it was present and mainly located in the middle and connecting piece. Moreover, we evaluated whether PKD would be involved in sperm-capacitation process. When mouse-spermatozoa were incubated under capacitating conditions with specific PKD inhibitors (Kb142-70 [5 uM] and CRT0066101 [2.5 uM]), an increase in events associated with capacitation such us tyrosine-phosphorylation and phospho-PKA substrates proteins was observed (WB). Also, an increase in motile (p<0.01), progressive (p<0.05) and hyperactivated (p<0.01; n=6) sperm population was observed (SCA, Micropticsystem). Additionally, the induction of the acrosomal reaction with progesterone -another event associated with capacitation- was evaluated using Acrosin-eGFP transgenic mouse sperm by flow cytometry. The inhibition of PKD increased the percentage of reacted spermatozoa (p<0.05;n=7). These findings allow us to conclude for first time that PKDs is present and active in mammalian sperm, and it would be involved in the regulation of the sperm capacitation process. Información suministrada por el agente en SIGEVAPalabras Clave
SPERMPKDCAPACITATION