title: Immature Oocytes from Unprimed Juvenile Mice Become a Valuable Source for Embryo Production When Using C-Type Natriuretic Peptide as Essential Component of Culture Medium.
abstract: C-type natriuretic peptide (CNP) and its receptor natriuretic peptide receptor 2 (NPR2) play a paramount role in the maintenance of oocyte meiotic arrest in antral follicles via the regulation of the intra-oocyte levels of cyclic guanosine monophosphate and cyclic adenosine monophosphate. We investigated the potential of CNP 1) to maintain oocyte meiotic arrest during a prolonged prematuration culture and 2) to sustain acquisition of developmental competence of immature cumulus-oocyte complexes (COCs). Compact COCs were collected from small antral follicles of prepubertal unprimed mice and placed in prematuration culture under different CNP-supplemented media conditions. A preliminary analysis showed a dose-dependent effect of CNP on the maintenance of meiotic arrest. A dose of 25 nM maintained oocytes under meiotic arrest for 24 h, and this period was extended to 48 h in the presence of estradiol. Analysis of transzonal projections of COCs cultured with CNP indicated that oocyte-cumulus connections were well preserved after the prolonged prematuration culture. Furthermore, CNP medium supplemented with FSH and GDF9 promoted oocyte growth and induced a shift in oocyte chromatin configuration from a predominantly dispersed to a condensed configuration. Following in vitro maturation, oocytes cultured under CNP were capable of extruding the first polar body at a high rate (around 80%). Blastocyst formation was significantly improved when oocytes were cultured under CNP-supplemented medium containing FSH and GDF9. This study reports for the first time a prolonged prematuration culture system, with CNP as the pivotal factor, that can efficiently maintain oocytes retrieved from unprimed prepubertal mice under meiotic arrest while promoting their acquisition of developmental competence.
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Sergio
authors.initials: S
authors.lastname: Romero
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Flor
authors.initials: F
authors.lastname: Sánchez
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Francesca
authors.initials: F
authors.lastname: Lolicato
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Heidi
authors.initials: H
authors.lastname: Van Ranst
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium johan.smitz@uzbrussel.be.
authors.forename: Johan
authors.initials: J
authors.lastname: Smitz
keywords: ART; CNP; IVF; IVM; embryo; meiosis; oocyte; oocyte maturation