Neuronal Gonadotrophin-Releasing Hormone (GnRH) and Astrocytic Gonadotrophin Inhibitory Hormone (GnIH) Immunoreactivity in the Adult Rat Hippocampus.

Gonadotrophin-releasing hormone (GnRH) and gonadotrophin inhibitory hormone (GnIH) are neuropeptides secreted by the hypothalamus that regulate reproduction. GnRH receptors are not only present in the anterior pituitary, but also are abundantly expressed in the hippocampus of rats, suggesting that GnRH regulates hippocampal function. GnIH inhibits pituitary gonadotrophin secretion and is also expressed in the hippocampus of a songbird; its role outside of the reproductive axis is not well established. In the present study, we employed immunohistochemistry to examine three forms of GnRH [mammalian GnRH-I (mGnRH-I), chicken GnRH-II (cGnRH-II) and lamprey GnRH-III (lGnRH-III)] and GnIH in the adult rat hippocampus. No mGnRH-I and cGnRH-II+ cell bodies were present in the hippocampus. Sparse mGnRH-I and cGnRH-II+ fibres were present within the CA1 and CA3 fields of the hippocampus, along the hippocampal fissure, and within the hilus of the dentate gyrus. No lGnRH-III was present in the rodent hippocampus. GnIH-immunoreactivity was present in the hippocampus in cell bodies that resembled astrocytes. Males had more GnIH+ cells in the hilus of the dentate gyrus than females. To confirm the GnIH+ cell body phenotype, we performed double-label immunofluorescence against GnIH, glial fibrillary acidic protein (GFAP) and NeuN. Immunofluorescence revealed that all GnIH+ cell bodies in the hippocampus also contained GFAP, a marker of astrocytes. Taken together, these data suggest that GnRH does not reach GnRH receptors in the rat hippocampus primarily via synaptic release. By contrast, GnIH might be synthesised locally in the rat hippocampus by astrocytes. These data shed light on the sites of action and possible functions of GnRH and GnIH outside of the hypothalamic-pituitary-gonadal axis.

The oxygenase reaction of acetolactate synthase.

In addition to the physiological reactions catalyzed by acetolactate synthase, it supports an oxygen-consuming side reaction. Although the synthase and oxygenase activities are activated to somewhat different extents by various metals (Mn2+, Mg2+, Ca2+, Co2+, Zn2+, Ni2+, Cd2+, Cu2+, Ba2+, Al3+), the modest degree of these differences (at most 6-fold) and the high degree of promiscuity of the enzyme with respect to its metal requirement suggest that the metal is not intimately involved in the chemistry of either reaction. Saturation of the oxygenase reaction occurs at pyruvate concentrations below the limit of sensitivity for the oxygen electrode (< 10 microM), at higher concentrations pyruvate inhibits the rate of oxygen consumption. At a noninhibitory concentration of pyruvate (1 mM), inhibition of the reaction is also observed with alpha-ketobutyrate. Inhibition of the oxygenase reaction by high concentrations of pyruvate or alpha-ketobutyrate is presumably due to competition between these substrates and molecular oxygen for a common carbanionic reaction intermediate, the conjugate base of (hydroxyethyl)thiamin pyrophosphate. Inhibition of the reaction indicates that the lactylthiamin pyrophosphate intermediate can decarboxylate prior to binding of the second pyruvate or alpha-ketobutyrate. At high concentrations of pyruvate or alpha-ketobutyrate, only incomplete inhibition of the oxygenase reaction is achieved (65-89% or 89-93% maximal inhibition, respectively). This incomplete inhibition of the oxygenase reaction by alpha-keto acids indicates that the reaction is not Theorell-Chance with respect to addition of the second alpha-keto acid and that oxygen has more than one route of access to the carbanionic reaction intermediate.