The main aim of this study was the observation of changes in two sperm proteins that are relationated with both the spatial distribution and the functional control of mitochondria, actin and mitofusin2. For this purpose, boar sperm from fresh ejaculates were subjected to an ‘in vitro’ capacitation (IVC) and further progesterone-induced, acrosome reaction (IVAR) by incubation at 39_C in a 5% CO2 atmosphere for 4 h in an specific capacitation medium. Sperm aliquots were taken at 0 h and 4 h of IVC and 5 min and 60 min after acrosome reaction. Our results show the presence of mitofusin2 in sperm for the first time, both through Western blot and immunofluorescence analyses. Regarding the overall expression, actin did not undergo changes during all the incubation period. Mitofusin2 progressively diminished after IVAR. Actin immunocytochemistry showed that a subequatorial band appeared after 4 h of IVC and after 60 min of IVAR the connecting piece showed a strong increase in fluorescence. Moreover, IVC and IVAR induced specific changes in the mitofusin2 location into the midpiece. We can conclude that IVC and IVAR produce changes in these two proteins, what would be related to the regulation of mitochondrial activity during these processes.
MITOFUSIN2 AND ACTIN DETERMINATION AND LOCALIZATION IN BOAR SPERMATOZOA DURING IN VITRO CAPACITATION AND FURTHER ACROSOME REACTION
BUCCI, DIEGO;
2009
Abstract
The main aim of this study was the observation of changes in two sperm proteins that are relationated with both the spatial distribution and the functional control of mitochondria, actin and mitofusin2. For this purpose, boar sperm from fresh ejaculates were subjected to an ‘in vitro’ capacitation (IVC) and further progesterone-induced, acrosome reaction (IVAR) by incubation at 39_C in a 5% CO2 atmosphere for 4 h in an specific capacitation medium. Sperm aliquots were taken at 0 h and 4 h of IVC and 5 min and 60 min after acrosome reaction. Our results show the presence of mitofusin2 in sperm for the first time, both through Western blot and immunofluorescence analyses. Regarding the overall expression, actin did not undergo changes during all the incubation period. Mitofusin2 progressively diminished after IVAR. Actin immunocytochemistry showed that a subequatorial band appeared after 4 h of IVC and after 60 min of IVAR the connecting piece showed a strong increase in fluorescence. Moreover, IVC and IVAR induced specific changes in the mitofusin2 location into the midpiece. We can conclude that IVC and IVAR produce changes in these two proteins, what would be related to the regulation of mitochondrial activity during these processes.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
