We have developed and validated a push-pull technique that allows focal perfusion of the ovary in unanesthetized freely moving rats. We have used this method to investigate the intraovarian secretion of catecholamines (dopamine, norepinephrine, epinephrine), oxytocin, beta-endorphin and gamma-amino-butyric acid (GABA) during the estrous cycle. Cycling animals were implanted with ovarian push-pull catheters and jugular vein catheters under ether anaesthesia on proestrus, estrus and diestrous Day 2. This procedure did not disrupt normal preovulatory release of prolactin and luteinizing hormone (LH). Thus, perfusion of the ovary and simultaneous monitoring of hormone levels in systemic blood in freely moving rats allow correlation of the temporal relationship of ovarian events with cyclic gonadotropin secretion. The results clearly indicate that a rise in ovarian norepinephrine occurs concomitant with the preovulatory surge in prolactin and LH. Ovarian beta-endorphin concentrations exhibit cyclic changes, whereas GABA release rates remain stable throughout the cycle. Oxytocin is secreted by ovarian tissue, and the secretion rate appears to be inversely related to prolactin. In view of the proposed involvement of ovarian nerves and particularly catecholamines in the process of follicular maturation and ovulation, our findings suggest a preovulatory activation of ovarian noradrenergic sympathetic neurons.
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