Preovulatory rise of NGF in ovine follicular fluid: possible involvement in the control of oocyte maturation.
Since nerve growth factor (NGF) is produced in vitro by granulosa cells after gonadotropin stimulation, the present research has been designed to investigate whether this neurotropin is involved in the events triggered by the gonadotropin surge that lead the follicle to ovulate a mature oocyte. To this aim, NGF levels in follicular fluid, collected before or 20 hours after the gonadotropin surge, was measured by ELISA. To evaluate whether NGF may have a non-neurotropic effect on follicle cells, the presence of NGF receptors was investigated by immunohistochemistry and further evaluated by analysing the tyrosine-phosphorylation pattern after NGF stimulation in vitro. The effect of NGF on the degree of cumulus expansion, cumulus-oocyte metabolic coupling, and meiotic maturation was finally studied by using the culture of follicle-enclosed oocyte. The results demonstrate that GnRH causes a dramatic rise of NGF in large follicles. Immunohistochemistry revealed a discrete positivity for trkA receptors localised in cumulus cells. Tyrosine phosphorylation pattern confirms that somatic cells are capable to transduce NGF signal. By contrast, all the oocytes examined were negative for trkA and did not change the phosphorylation pattern after NGF. In vitro NGF (100 ng/ml) induced a marked cumulus expansion and a progressive cumulus-oocyte uncoupling similar to that produced by gonadotropins. The addition of NGF also caused the resumption of meiosis in more than 70% of the oocytes analysed with an effect that is only slightly less pronounced than that of gonadotropins (80%). The increase in NGF secretion following gonadotropin surge suggests that this neurotropin may be involved in the control of oocyte maturation.
Barboni
Barbara
B
Dipartimento di Strutture, Funzioni e Patologie Animali e Biotecnologie, Fisiologia Veterinaria, Università di Teramo, 64100 Italy. barboni@ifv.vet.unite.it
Mattioli
Mauro
M
Gioia
Luisa
L
Turriani
Maura
M
Capacchietti
Giulia
G
Berardinelli
Paolo
P
Bernabò
Nicola
N
D000818:Animals; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015571:Follicular Fluid / Q000378:metabolism*; D066298:In Vitro Techniques; D008540:Meiosis / Q000187:drug effects; D009414:Nerve Growth Factors / Q000378:metabolism / Q000502:physiology*; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development / Q000502:physiology*; D012756:Sheep
D006063:Chorionic Gonadotropin; D009414:Nerve Growth Factors