INVESTIGADORES
MAYORGA Luis Segundo
congresos y reuniones científicas
Título:
Permeant proteins as a tool to study cortical granules exocytosis in mouse oocytes.
Autor/es:
GALLO, GL; DE PAOLA, MM; RODRÍGUEZ PEÑA, M; MAYORGA LS; MICHAUT, M.A.
Lugar:
Mendoza
Reunión:
Congreso; 48º SAIB; 2012
Institución organizadora:
SAIB
Resumen:
One of the most important events that are produced in oocytes as a
result of fertilization is cortical granule exocytosis (CGE), a
secretory process that produces polispermy blockade and ensures
normal embryonic development. Nevertheless, the molecular
mechanism of this process remains unknown. Previous results in
our laboratory show that MARCKS, a prominent substrate of PKC
involved in exocytosis in different cell types, participates in CGE in
mouse oocyte. In this cellular model, proteins are introduced by
microinjection, which is a very valuable and useful technique, but it
causes an injury to the cells which is reflected in high mortality and
low yields. We decided to tune up the usefulness of permeant
proteins to study exocytotic mechanisms in mouse oocyte. We used
the permeant peptide corresponding to MARCKS effector domain
(amino acids 154-165) conjugated to thrimethylrhodamine (TMRED). Oocytes were incubated with increasing concentrations of this
peptide, previous remotion of zona pellucida; then cells were
parthenogenetically activated with SrCl2, and cortical granules
were quantified with the program Image J. TMR-DE was able to
inhibit CGE and this effect was concentration-dependent. These
results corroborate previous data obtained in our laboratory by
protein microinjection, and show the efficiency of permeant
proteins as a tool to study CGE in moouse oocytes.