Abstract
Investigations in the author's laboratory which bear upon several fundamental aspects of the mammalian neurohypophysis were reviewed and related to well-established or currently accepted concepts, (a) The separation of vasopressin-rich from oxytocin-rich neurosecretory granules (NSG) from posterior pituitary homogenates was achieved. (b) The hormone and amino acid composition of isolated NSG was found to be almost identical to that of the van Dyke protein, prepared by chemical extraction of posterior pituitary glands, indicating that the protein is synonymous with the NSG carrier protein to which vasopressin (VP) and oxytocin (OT) are bound. (c) Pinched-off nerve terminals (neurosecretosomes) were isolated from homogenates by centrifugation and shown to be metabolically and morphologically intact. The hexosemonophosphate shunt was highly active in the nerve terminals, where the peptide hormones are stored and secreted, but not in the cell bodies within hypothalamic nuclei, where they are synthesized. (d) Neurosecretosomes relatively enriched in VP were separated from those rich in OT by density-gradient centrifugation, providing support for the concept that a given neurohypophyseal neuron stores and secretes only one of the hormones. (e) Electron microscopy indicated that microvesicles, about 50 mμ in diameter, and called "synaptic" vesicles by some workers, are derived from membranes of depleted NSG and are not of the type that contain neurotransmitter. (f) Acetylcholine (ACh), choline acetyltransferase, and acetylcholinesterase were identified in the posterior pituitary gland and are present in about the same proportions but 1/5 to 1/10 the activity as in whole brain. ACh was found in the neurosecretosome and microvesicle centrifugation fractions. The three cholinergic components in the posterior pituitary are believed to be constituents, not of neurosecretory nerve endings which contain VP or OT, but of specific cholinergic neurons whose role in neurohypophyseal physiology remains to be elucidated. (g) VP and OT are generally released together from the gland in vivo and from posterior pituitary tissue or NSG in vitro. However, exceptions can occur both in vivo and in vitro in which one of the hormones is apparently released exclusively. Studies on the release of VP and OT in vitro show that the NSG–hormone complex is more labile in the case of OT than of VP, an observation that suggests a molecular basis for numerous in vivo findings in which a preponderance of OT over VP is secreted in response to a wide variety of stimuli.
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