Central obestatin administration affect the LH and FSH secretory activity in peripubertal sheep.

Obestatin - a 23 amino acid peptide is synthesized as another product of the ghrl gene and its synthesis occurs mainly in gastric mucosa cells. This hormone is involved in complex gut-brain neurohormonal networks, thereby can participates in the modulation of gonadotrophic axis activity. The aim of this study was to investigate the consequence of intracerebroventricular infusions of obestatin on LH and FSH pituitary cells secretory activity in peripubertal female sheep. Animals were randomly divided into two groups: the control group (n = 14) received intracerebroventricular infusions of Ringer-Lock solution (120 μL h), and the obestatin group (n = 14) was infused with obestatin (25 μg/120 μL h) diluted in Ringer-Lock solution. A series of four infusions was performed on three consecutive days. Blood samples were collected on day 0 and day 3. The sheep were slaughtered immediately after the end of the experiment. For molecular biological analysis, pituitaries from 7 sheep from each group (n = 7 + 7) were prepared and frozen in liquid nitrogen immediately after collection and then stored at -80 °C until Real Time RT-qPCR and RIA analyzes. For immunohistochemical analysis, pituitary tissues from the remaining animals (n = 7 + 7) was fixed in situ for further examination. Real-Time qPCR and immunohistochemistry analyses revealed substantial changes in the LH and FSH pituitary cells secretory activity in obestatin-infused sheep. Exogenous obestatin administration reduced LHβ mRNA expression and increased the accumulation of immunoreactive LH in gonadotrophic cells of the adenohypophysis. These changes were accompanied by a decrease in the mean LH concentration in the peripheral blood resulting from the lower LH pulse amplitude. Moreover, an increase in both FSHβ mRNA expression and FSH immunoreactivity and amount in pituitary cells were noted, while mean blood FSH concentration remained unchanged after obestatin treatment. The obtained results showed that exogenous obestatin affected LH secretory activity at the level of protein synthesis, accumulation and release as well as obestatin increase FSHβ mRNA expression and accumulation of this hormone but at the same time have no effect on FSH release to blood. Thus, obestatin can participate in the neuroendocrine network, which modulates gonadotrophic axis activity in sheep.