Immunolesion of melanopsin neurons causes gonadal regression in Pekin drakes (Anas platyrhynchos domesticus)

Abstract

Several light sensitive receptors have been described in the avian brain that are thought to regulate the reproductive axis independently from the eyes and pineal gland. Recently, our lab has described the presence of three of these photoneuroendocrine systems in the Pekin duck: opsin, opsin 5, & melanopsin. We set out to test the hypothesis that melanopsin receptive neurons are necessary to maintain seasonal reproductive status along with growth and development in the Pekin drake. To accomplish these goals we first investigated 50-week-old Pekin drakes that were housed in the aviary at Hope College under long day length (18h lights on) conditions in floor pens. To specifically lesion melanopsin-receptive neurons, 3μl of an anti-melanopsin-saporin conjugate (MSAP, 100ng/ul) was injected into the lateral ventricle (n=10). Control drakes were injected with 3μl of equimolar unconjugated anti-melanopsin and saporin (SAP, n=10). Reproductive behaviors were analyzed weekly in a test pen with adult hens and MSAP drakes showed a significant (p<0.01) reduction in reproductive behaviors after week 2. After 5weeks, drakes were euthanized and body weights were measured, and brains, pituitaries, and testes collected and stored for analyses. Mature MSAP-treated drakes had significantly (p<0.001) reduced relative teste weights compared to SAP controls. qRT-PCR analyses of hypothalamus showed a significant reduction (p<0.001) in GnRH and melanopsin mRNA levels, but not opsin 5, vertebrate ancient opsin, or opsin 2 (rhodopsin). Immunocytochemical analyses showed a significant reduction (p<0.01) in tyrosine hydroxylase-immunoreactivity in the PMM. These data suggest that although blue light alone is not able to maintain testicular function, the blue-light sensitive melanopsin activity is critical to maintain gonadal function.