pituitrin and pregnenolone-sulfate

The neurosteroid pregnenolone sulphate (PS) interacts allosterically with ionotropic glutamate receptors and thereby could be an important modulator of activity within the hypothalamic magnocellular nuclei. The present in-vitro study therefore examined the effect of perifusion of PS (100 microM) on activity of supraoptic oxytocin (OT) and vasopressin (VP) neurones, in which firing was stimulated by local application of glutamate, NMDA or AMPA. In the presence of locally applied glutamate, PS significantly potentiated firing in putative VP neurones, but had little effect on putative OT neurones. In both cell types, PS increased firing in the presence of NMDA and depressed firing in the presence of AMPA. The action of PS on glutamate- and NMDA-stimulated firing was unaffected by addition of the GABA(A) receptor antagonist, picrotoxin (50 microM). The suppressive action of PS on AMPA-stimulated firing was, however, reversed by picrotoxin and therefore probably requires intact GABAergic transmission for its expression. When putative VP neurones were stimulated by local application of K+, in the presence of picrotoxin, PS evoked a small increase in the ongoing activity, although this did not reach significance. When the glutamate receptor antagonists, NBQX (20 microM) and AP5 (40 microM), were included in the medium, no change in K+ -stimulated firing was observed. Hence PS has no effect on activity of putative VP neurones in the absence of exogenous and endogenous glutamate excitation. In conclusion, PS selectively potentiates glutamate-stimulated activity in putative VP neurones, probably via NMDA receptors, thus providing a mechanism whereby this neurosteroid might exert rapid non-genomic effects on VP secretion. The lack of effect of PS in putative OT neurones probably relates to the relatively small involvement of NMDA receptors in mediating glutamate excitation in this cell type.

Topics: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Animals; Electrophysiology; Excitatory Amino Acid Agonists; Female; Glutamic Acid; N-Methylaspartate; Neurons; Oxytocin; Pregnenolone; Rats; Rats, Wistar; Supraoptic Nucleus; Vasopressins

Topics: Animals; Evoked Potentials; Female; In Vitro Techniques; Neurons; Oxytocin; Pregnenolone; Rats; Supraoptic Nucleus; Vasopressins

The effect of the neurosteroid pregnenolone sulphate (PS) on N-methyl-D-aspartate (NMDA)-induced phasic firing of supraoptic vasopressin (VP) neurones was studied in rat hypothalamic slices in vitro. In VP neurones which were induced to fire phasically by continuous perifusion with NMDA (9-30 microM), addition of 100 microM PS to the incubation medium significantly increased overall spike frequency, with a rise in both proportion of time active and intraburst firing rate. A similar effect was seen during picrotoxin block of GABAergic transmission. No significant change in NMDA-induced phasic firing was observed with 100 microM dehydroepiandrosterone sulphate. VP neurones became silent in the absence of NMDA, and under these conditions PS had no effect. In conclusion, PS increases NMDA-induced phasic firing in VP neurones, providing a mechanism whereby this neurosteroid may participate in the regulation of VP secretion.

Topics: Animals; Dehydroepiandrosterone Sulfate; Drug Synergism; Evoked Potentials; Female; Hypothalamus; In Vitro Techniques; N-Methylaspartate; Neurons; Picrotoxin; Pregnenolone; Rats; Rats, Wistar; Supraoptic Nucleus; Time Factors; Vasopressins