Effect of gonadotropins on oocyte maturation in vitro: an animal model.
OBJECTIVE
Analysis of the effects of human-derived gonadotropin drugs, FSH and LH (Repronex) and hCG (Novarel), on oocyte maturation, using a porcine oocyte in vitro maturation system as a culture model.


METHODS
Randomized research experimental study.


METHODS
Academic basic research laboratory.


METHODS
Prepubertal gilts that were slaughtered in the local slaughter house.


METHODS
Oocytes will be exposed to immunofluorescent staining and confocal laser scanning microscopy: Western blot analysis on cumulus-oocyte-complexes following treatment with different concentrations of the gonadotropin drugs Repronex, Novarel, and a Repronex and Novarel combination.


METHODS
Analysis of porcine oocyte spindle and chromosomal configuration with alpha-tubulin-fluorescein isothiocyanate antibody and propidium iodide staining. Porcine oocyte mitochondrial distribution and aggregation pattern staining was assessed with Mito Tracker Red CMXRox probe. Porcine oocyte cortical granule distribution was observed via peanut agglutinin-fluorescein isothiocyannate staining; Western blot analysis detected extra-cellular signal-regulated kinase 1/2 activation in cumulus cells.


RESULTS
An increase of gonadotropin concentration in the culture medium resulted in an increase in the following: the percentage of oocytes reaching metaphase II, normal configuration of the spindle, normal chromosomal alignment, cortical granule migration, and mitochondrial aggregation. Levels of nuclear and cytoplasmic maturation peaked as the concentration of gonadotropins approached its threshold level.


CONCLUSIONS
Addition of a threshold concentration of the gonadotropin drugs Repronex, Novarel, and a combination of the two can significantly improve porcine oocyte maturation in vitro.
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.
Wei
W
Sha
Bao-Zeng
BZ
Xu
Mo
M
Li
Di
D
Liu
Huai L
HL
Feng
Qing-Yuan
QY
Sun
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D020090:Chromosome Segregation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004359:Drug Therapy, Combination; D005260:Female; D005300:Fertility Agents, Female / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D043373:Follicle Stimulating Hormone, Human / Q000494:pharmacology*; D006801:Humans; D007986:Luteinizing Hormone / Q000494:pharmacology*; D008677:Metaphase / Q000187:drug effects; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D023421:Models, Animal; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D009930:Organic Chemicals / Q000494:pharmacology; D010766:Phosphorylation; D008941:Spindle Apparatus / Q000187:drug effects; D013552:Swine
D006063:Chorionic Gonadotropin; D004338:Drug Combinations; D005300:Fertility Agents, Female; D043373:Follicle Stimulating Hormone, Human; D009930:Organic Chemicals; C432248:Repronex; D007986:Luteinizing Hormone; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3