Nitric oxide (NO) is generated from L-arginine by different isoforms of the enzyme nitric oxide synthase (NOS) and is known to be involved in mediating several biological functions, some of which are associated with reproduction. In this study, we examined the ability of the prepubertal ovary to express the inducible (i), as well as the neuronal-type constitutive (c) form of NOS and also, investigated whether either isoform undergoes changes in the ovary during peripubertal development. Results indicate that both forms of NOS were expressed in the ovary and that the iNOS mRNA transcripts were expressed without being exogenously induced. When compared with juvenile levels, iNOS, but not cNOS, mRNA increased (p<0.01) during the early proestrous phase of development. By the late proestrous phase, the levels of iNOS mRNA declined markedly (p<0.001) and remained low throughout both the first estrous and diestrous phases. Western blot analysis revealed both iNOS and cNOS protein expression in each phase of puberty with only iNOS showing a significant change during the peripubertal period. Specifically, there was an initial increase in the expression of iNOS protein during the late proestrous phase (p<0.05) which was accompanied by preovulatory increases in serum estradiol (p<0.01) and LH (p<0.001). The iNOS protein levels then dramatically increased to peak on the morning of first estrus (p<0.001), an event associated with declining (p<0.05) serum levels of estradiol. These data demonstrate developmental changes in the expression of ovarian iNOS mRNA and protein both before and after first ovulation; hence, suggesting a role for NO in the ovary during pubertal maturation and furthermore, providing compelling evidence at the gene level supporting the hypothesis that the NO/NOS system plays a physiological role in ovarian function.
