Microfilament disruption is required for enucleation and nuclear transfer in germinal vesicle but not metaphase II human oocytes.
OBJECTIVE
To evaluate the usefulness of microfilament disruption before enucleation and nuclear transfer in human oocytes at different stages of maturation.


METHODS
Prospective experimental study.


METHODS
Private clinics.


METHODS
Infertile couples undergoing assisted reproduction attempts.


METHODS
Oocyte enucleation and nuclear transfer, activation of reconstructed oocytes.


METHODS
Oocyte survival, nuclear transfer efficacy, activation outcomes.


RESULTS
Survival rate and nuclear transfer efficacy of germinal vesicle oocytes exposed to the microfilament disrupting agent cytochalasin B before enucleation were 88% and 80%, respectively. These figures dropped, respectively, to 8% and 2% when cytochalasin treatment was omitted. By contrast, cytochalasin-treated and -untreated metaphase II oocytes showed similar survival rate (87% vs. 90%) and nuclear transfer efficacy (78% vs. 87%). This also applied to metaphase II oocytes matured in vitro from the germinal vesicle stage. Cytochalasin treatment did not affect activation rate of reconstructed oocytes, but it increased the occurrence of oocytes with multiple female pronuclei.


CONCLUSIONS
Microfilament disruption before enucleation is required for germinal vesicle oocytes but not for metaphase II oocytes.
Tesarik
Jan
J
MAR&Gen, Molecular Assisted Reproduction and Genetics, Granada, Spain. cmendoza@ugr.es
Martinez
Francisco
F
Rienzi
Laura
L
Ubaldi
Filippo
F
Iacobelli
Marcello
M
Mendoza
Carmen
C
Greco
Ermanno
E
D008841:Actin Cytoskeleton / Q000502:physiology*; D000001:Calcimycin / Q000494:pharmacology; D002467:Cell Nucleus / Q000502:physiology; D003571:Cytochalasin B / Q000494:pharmacology; D005260:Female; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008677:Metaphase / Q000502:physiology; D008846:Micromanipulation / Q000379:methods*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000502:physiology*; D011446:Prospective Studies
D007476:Ionophores; D000001:Calcimycin; D003571:Cytochalasin B