The hypothalamo-hypophysial system of the lamprey,Lampetra fluviatilis L.

The distribution of monoaminergic structures was studied in the proximal neurosecretory contact region and neurohypophysis of the lamprey by light and electron microscopic radioautography. Only weak radioautographic reactions were found in the proximal neurosecretory contact region 1 h after injection of 3H-dopamine. High-resolution radioautography revealed some labeled neurosecretory terminals mainly in contact with the basement membrane of the connective tissue layer separating the proximal neurosecretory contact region from the hypophysial pars distalis. The number of silver grains as well as the number of neurosecretory terminals marked by the presence of labeled dopamine was much higher in the neurohypophysis of the same species. In the latter, labeled neurosecretory terminals were found in contact with the connective tissue layer containing blood vessels of the general circulation. Some neurosecretory terminals make synaptoid contacts with tanycyte perikarya and their basal processes. According to their ultrastructure and the size of their granules, the labeled neurosecretory terminals are identical with the B type terminals described in both neurohemal regions (transmission electron microscopy). No labeled neurosecretory terminals were observed in the proximal neurosecretory contact region and the neurohypophysis of lampreys treated with the serotonin precursor, 3H-5-hydroxytryptophan.     

Cytochemical ultrastructural study of the glycoproteins in the rat hypothalamo-post-pituitary complex]

The results obtained with various methods applied to the cytochemical detection of carbohydrates at an ultrastructural level, confirm the existence of glycoproteins in neurosecretory material in the neurohypophysis as well as in the hypothalamic magnocellular nuclei. This glycoproteic component, however, is not present in all the secretory granules and, according to their cytochemical behaviour, it is possible to distinguish two types of neurosecretory fibres: one where all the granules respond negatively; the other where most of the granules are reactive. The existence of two types of neurons corresponding to these two fibres cannot yet be asserted, but seems very likely, perhaps connected with the hormonal duality of the magnocellular nuclei. The reactions are also positive on the Golgi apparatus, in accordance with its function in glycoprotein synthesis. But the difference of reactivity between the Golgi cisternae and the neurosecretory product suggests that glycoprotein synthesis is still going on in the neurosecretory granules outside the Golgi area.     

Plasticity in the hypothalamic magnocellular neurosecretory system

Over the past decade or so, plasticity has emerged as an important, quantifiable property of the mammalian hypothalamic magnocellular neurosecretory system. This plasticity has turned out to be genuinely related to normal function in the sense that it is a set of responses to physiological stimulation rather than only the sequelae of insult or injury, and it is generally completely reversible. This latter property, of course, distinguishes it further from the plasticity observed after injury. Four features of this magnocellular system that have been shown to display predictable and reversible intercellular plasticity are reviewed: the relationships between neurons and their associated astrocytic glia at various levels (dendritic somatic and terminal) of the magnocellular elements; the extent of terminal and glial contact with the basement lamina in the neurohypophysis; the type and possible efficacy of synaptic input, and the extent of electrotonic coupling among the magnocellular neurons.     

Histochemical studies on the neurosecretory system of the common garden lizard Calotes versicolor (Daudin) under experimental condition

Summary Histochemical localizations of various substances in the hypothalamic neuro-secretory cells as well as neurohypophysis of the normal and stressed (scalded) garden lizards, Calotes versicolor suggest that the neurosecretory materials, secreted by the hypothalamic neurones may be a mucoprotein-lipoprotein complex which is finally stored in the neuro-hypophysis. Only the proteinaceous fraction of this complex appears to be released by the nerve terminals in the neurohypophysis under the stressed condition. Such suggestion implies a chemical transformation of neurosecretory substances after leaving the neurosecretory cell body.     

The hypothalamo-hypophysial system in acipenseridae

Summary The percentage of peptidergic (A₁ and A₂) and adrenergic (B) neurosecretory terminals was studied in the neurohypophysis of sexually mature female sturgeons. Neurosecretory terminals of the A₂ type prevail in the neurohypophysis, whereas A₁ and B terminals are rare. The activity of these types of terminals was established (1) during upstream migration, (2) shortly after spawning, and (3) three to six weeks after spawning. Terminals of B type are the most active elements during all the periods studied. These elements become strongly activated in sturgeons during upstream migration, i.e., earlier than the peptidergic neurosecretory terminals. Peptidergic terminals, especially elements of type A₂, become synchronously and strongly activated in fish shortly after spawning. In the late postspawning period neurosecretory terminals of all three types become synchronously inactive, persisting in a quiescent state in comparison to the two previous periods. The appearance of neurosecretory material discharged into the intercellular clefts by exocytosis correlates on the whole with the activity level of the A₁ and A₂ terminals in each individual studied. A functional correlation exists between the activity of the peptidergic and adrenergic neurosecretory terminals in the neurohypophysis. The data obtained are discussed with reference to a concept regarding spawning in some fish species as a physiological stress (Polenov et al., 1976). A possible dual control (peptide and monoamine neurohormones) over the function of visceral organs and glandular cells of the intermediate lobe of the hypophysis is also suggested (Polenov, 1970, 1975, 1978; Polenov and Belenky, 1973).Dedicated to the memory of Professor Wolfgang Bargmann, a great scientist and a generous friend     

Corticotrophic cells in the rostral zone of the pars intermedia and in the adjacent neurohypophysis of the rat and mouse

Summary In the mouse, the rostral zone of the pars intermedia is almost exclusively composed of typical corticotrophic cells. They are located around and even within the neural stalk, at the level of transition between stalk and neural lobe. In the rat, the corticotrophic cells of the rostral zone are found in scattered islets among the MSH producing cells, and also in the neural lobe. In both the rat and mouse, these cells are in direct contact with various types of nerve terminals. Synaptoid contacts with aminergic and neurosecretory nerve fibers are observed. Furthermore they are also closely related to the hypophysial portal vessels. Following adrenalectomy, the cells located in the neurohypophysis always react more intensely than tose in the rostral zone. The functional significance of these corticotrophic cells which are subject to both humoral and neural regulation remains as yet hypothetical. Their participation in neurogenic stress response seems probable.     

Multifaceted purinergic regulation of stimulus-secretion coupling in the neurohypophysis

The neurohypophysis is an original model of the CNS secretory system releasing vasopressin (AVP) and oxytocin (OXT), two neuropeptides hormones synthesized by the magnocellular neurons of the hypothalamus. Specific patterns of action potentials originating from cellular bodies of magnocellular neurons control the release of AVP and OT, but intra-neurohypophysis regulations do modulate the neuropeptides release. There is now good evidence for the effects of extracellular purines in the control of neurohypophysial secretion. This paper brings together evidence for the multiple, intricate actions of purines in the extracellular space of the neurohypophysis. It covers four main points. First, the activity-dependent release of endogenous ATP in the neurohypophysis. Second, the action of ATP on both neuronal and non-neuronal compartments of the neural lobe. Third, the termination of ATP positive feedback by ecto-nucleotidases. And finally the possible involvement of adenosine in the regulation of neurohypophysial secretion and glial plasticity. The data suggest that ATP and adenosine are physiological modulators of the release of neurohypophysial peptides by acting directly on nerve terminals and indirectly on neurohypophysial astrocytes. Since purinergic receptors are widespread in nervous and endocrine systems, the neurohypophysis appears as an useful model for studying the role of purines in the regulation of stimulus-secretion coupling and neuron-glia interactions. The feedback mechanisms found in the neurohypophysis could be ubiquitous, occurring throughout the central nervous system and in other secretory systems.     

FINE STRUCTURE OF THE NEUROHYPOPHYSIS OF THE OPOSSUM (DIDELPHIS VIRGINIANA)

The neurohypophysis of the opossum (Didelphis virginiana) was studied by electron microscopy in order to amplify Bodian''s classic light microscopic observations in which he demonstrated a definite lobular pattern. The lobule of the opossum neurohypophysis is divided into three regions: a hilar, a palisade, and a septal zone. The hilar portion contains bundles of nerve fibers, the extensions of the hypothalamo-hypophyseal tract containing neurofilaments but few neurosecretory granules. In the opossum, pituicytes have a densely fibrillar cytoplasm. Herring bodies are prominent in the hilar region. They are large bodies packed with neurosecretory granules that have been described as end bulb formations of axons. From the hilar region, axons fan out into a palisade zone where the nerve terminals packed with neurosecretory granules, mitochondria, and microvesicles abut upon basement membranes. The neurosecretory granules are similar to those present in the neurohypophysis of other mammals, except for an occasional huge granule of distinctive type. Material morphologically and histochemically resembling glycogen occurs as scattered particles and as aggregates within nerve fibers. The septal zone, containing collagen, fibroblasts, and numerous small capillaries, is separated from the adjacent glandular tissue by a basement membrane.     

Micromolar 4-aminopyridine enhances invasion of a vertebrate neurosecretory terminal arborization: optical recording of action potential propagation using an ultrafast photodiode-MOSFET camera and a photodiode array

Modulation of the amount of neuropeptide released from a neurosecretory tissue may be achieved by different means. These include alterations in the quantity secreted from each active nerve terminal or in the actual number of terminals activated. From the vertebrate hypothalamus, magnocellular neurons project their axons as bundles of fibers through the median eminence and infundibular stalk to arborize extensively and terminate in the neurohypophysis, where the neurohypophysial peptides and proteins are released into the circulation by a Ca-dependent mechanism. Elevating [Ca2+]o increases the magnitude of an intrinsic optical change in the neurohypophysial terminals that is intimately related to the quantity of neuropeptide released. Similarly, the addition of micromolar concentrations of 4-aminopyridine to the bathing solution enhances this change in large angle light scattering. However, we show here that, while these effects are superficially similar, they reflect different mechanisms of action. Evidence from intrinsic optical signals (light scattering) and extrinsic (potentiometric dye) absorption changes suggests that calcium increases the amount of neuropeptide released from each active terminal in the classical manner, while 4-aminopyridine exerts its secretagogue action by enhancing the invasion of action potentials into the magno-cellular neuron''s terminal arborization, increasing the actual number of terminals activated. Physiologically, electrical invasion of the complex terminal arborization in the neurohypophysis may represent an extremely sensitive control point for modulation of peptide secretion. This would be especially effective in a neurohaemal organ like the posterior pituitary, where, in contrast with a collection of presynaptic terminals, the precise location of release is less important than the quantity released.     

Radioautographic studies on the neurohypophysial projections of the supraoptic and paraventricular nuclei in the rat

Summary The distribution of labelled axonal pathways was studied after unilateral stereotaxic injection of ³H-leucine into either supraoptic (SON) or paraventricular nuclei (PVN). In addition to extrahypothalamic projections of both nuclei, the main efferents appeared to run towards the neurohypophysis, yet with a strikingly different pattern. At the neurohypophysial level, the SO-neurohypophysial tract crossed the inner layers of the median eminence (ME) before scattering in the neural lobe. The PV-neurohypophysial pathway, by contrast, provided an exclusive innervation to the external layer of the whole neurohypophysial organ, including the median eminence, infundibular stalk and neural lobe. The functional correlates of the clear-cut anatomical distinctness between the two magnocellular neurosecretory systems are discussed.     

Factors Governing Activity-Dependent Structural Plasticity of the Hypothalamoneurohypophysial System

1. The adult hypothalamoneurohypophysial system (HNS) undergoes reversible morphological changes in response to physiological stimulation.2. In the hypothalamus, stimulation of neurohormone secretion results in reducedastrocytic coverage of oxytocinergic somata and dendrites so that their surfaces becomedirectly juxtaposed. Concurrently, there is a significant increase in the number of GABAergic, glutamatergic, and noradrenergic synapses impinging on the neurons.3. In the neurohypophysis, stimulation induces retraction of pituicyte processes fromthe perivascular area and enlargement and multiplication of neurosecretory terminals.4. These neuronal-glial and synaptic changes are reversible with cessation of stimulation, thus rendering the HNS an excellent model to study physiologically linked structuralneuronal plasticity in the adult CNS.5. We still do not know the cellular mechanisms and factors underlying such plasticity.Recent studies indicate, however, that the adult HNS expresses molecular characteristicsnormally associated with histogenesis and/or tissue reorganization in developing or regenerating neural systems. They include expression of cell adhesion molecules such as the highlysialylated isoform of the neural cell adhesion molecule, PSA-NCAM, and the glycoproteins, F3 and tenascin-C.6. The expression of PSA-NCAM and tenascin-C does not show striking differencesin terms of age, sex or physiological condition but that of F3 varies considerably withneurohypophysial stimulation.7. We postulate that such molecular features allow magnocellular neurons and theirglia to undergo neuronal-glial and synaptic plasticity throughout life, provided the properstimulus intervenes.8. Thus, in the hypothalamic nuclei, centrally released oxytocin acting in synergy with steroids can induce such plasticity, while adrenaline, acting through -adrenergic mechanisms, does so in the neurohypophysis.     

Immuno-enzyme cytochemical demonstration of mesotocinergic nerve fibres in the pars intermedia of the amphibian hypophysis

Summary Immuno-enzyme cytochemical investigations showed that the whole amphibian pars intermedia of the hypophysis is innervated by an intercellular network of peptidergic varicose nerve fibres which contain mesotocin or (and) parts of the mesotocin molecule. The pars intermedia does not contain vasotocinergic fibres. The mesotocinergic fibres are branches of axons leaving the pituitary stalk and the neural lobe. In animals of which the hypothalamic magnocellular neurosecretory preoptic nuclei had been completely removed, the immuno-reactive mesotocinergic fibres of the pars intermedia had totally disappeared. From this result, it is concluded that the mesotocinergic fibres of the pars intermedia of the amphibian hypophysis are axons of neurosecretory perikarya located in the hypothalamic magnocellular neurosecretory preoptic nuclei.Dedicated to Professor Berta Scharrer on the accasion of her 70th birthdayThis investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Identification of adenohypophysiotropic neurohormone producing neurosecretory cells in Rana temporaria

Summary Electron microscopic investigation of neural isolated and normal pars ventralis of the tuber cinereum showed the presence, in Rana temporaria, of a tubero-hypophysial neurosecretory system. A striking structural resemblance between this parvicellular, aldehydefuchsin negative, tubero-hypophysial neurosecretory system and the magnocellular, aldehydefuchsin positive, hypothalamo-hypophysial neurosecretory system was observed. Six different neurosecretory cell types are described, characterized by different shape and size of their respective secretory granules. The nature of the secretory product of these cells is briefly discussed.Of the results of this investigation, a preliminary note has been published (Dierickx, 1971).The authors wish to thank Prof. Dr. H. Steyaert and Dr. W. Moerdijk of the Laboratory for Mathematical Statistics, Ghent University, for invaluable advice and assistance with their computers. They thank Dr. G. De Waele for his help with the scanning electron microscopy.     

The Ca2+ channel β2 subunit is selectively targeted to the axon terminals of supraoptic neurons

The assembly of high voltage-activated Ca²⁺ channels with different β subunits influences channel properties and possibly subcellular targeting. We studied β subunit expression in the somata and axon terminals of the magnocellular neurosecretory cells, which are located in the supraoptic nucleus (SON) and neurohypophysis, respectively. Antibodies directed against the 4 Ca_Vβ subunits (Ca_Vβ₁-Ca_Vβ₄) were used for immunoblots and for immunostaining of slices of these two tissues. We found that all 4 β subunits are expressed in both locations, but that Ca_Vβ₂ had the highest relative expression in the neurohypophysis. These data suggest that the Ca_Vβ₂ subunit is selectively targeted to axon terminals and may play a role in targeting and/or regulating the properties of Ca²⁺ channels.     

Calcium channels that are required for secretion from intact nerve terminals of vertebrates are sensitive to omega-conotoxin and relatively insensitive to dihydropyridines. Optical studies with and without voltage-sensitive dyes

Extrinsic absorption changes exhibited by potentiometric dyes have established the ionic basis of the action potential in synchronously activated populations of nerve terminals in the intact neurohypophyses of amphibia and mammals (Salzberg et al., 1983; Obaid et al., 1983, 1985b). Also, large and rapid changes in light scattering, measured as transparency, have been shown to follow membrane depolarization and to be intimately associated with the release of neuropeptides from the nerve terminals of the mouse neurohypophysis (Salzberg et al., 1985; Gainer et al., 1986). We report some experiments that help to define the pharmacological profile of the calcium channels present in intact neurosecretory terminals of vertebrates. For these, we used the peptide toxin omega-conotoxin GVIA (1-5 microM) and the dihydropyridine compounds Bay-K 8644 and nifedipine (2-5 microM), together with the after-hyperpolarization of the nerve terminal action potential. This undershoot depends upon the activation of a calcium-mediated potassium channel, as suggested by its sensitivity to [Ca++]o and charybdotoxin. omega-conotoxin GVIA substantially reduced the after-hyperpolarization in neurosecretory terminals of Xenopus, while neither of the dihydropyridine compounds had any effect under conditions that mimic natural stimulation. The effects of these calcium channel modifiers on the action potential recorded optically from the terminals of the Xenopus neurohypophysis were faithfully reflected in the behavior of the light-scattering changes observed in the neurohypophysis of the CD-1 mouse. omega-conotoxin GVIA (5 microM) reduced the size of the intrinsic optical signal associated with secretion by 50%, while the dihydropyridines had little effect. These observations suggest that the type of calcium channel that dominates the secretory behavior of intact vertebrate nerve terminals is at least partially blocked by omega-conotoxin GVIA and is insensitive, under normal conditions, to dihydropyridines.     

Microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of presynaptic nerve terminals

Nerve endings of the posterior pituitary are densely populated by dense- core neurosecretory granules which are the storage sites for peptide neurohormones. In addition, they contain numerous clear microvesicles which are the same size as small synaptic vesicles of typical presynaptic nerve terminals. Several of the major proteins of small synaptic vesicles of presynaptic nerve terminals are present at high concentration in the posterior pituitary. We have now investigated the subcellular localization of such proteins. By immunogold electron microscopy carried out on bovine neurohypophysis we have found that three of these proteins, synapsin I, Protein III, and synaptophysin (protein p38) were concentrated on microvesicles but were not detectable in the membranes of neurosecretory granules. In addition, we have studied the distribution of the same proteins and of the synaptic vesicle protein p65 in subcellular fractions of bovine posterior pituitaries obtained by sucrose density centrifugation. We have found that the intrinsic membrane proteins synaptophysin and p65 had an identical distribution and were restricted to low density fractions of the gradient which contained numerous clear microvesicles with a size range the same as that of small synaptic vesicles. The peripheral membrane proteins synapsin I and Protein III exhibited a broader distribution extending into the denser part of the gradient. However, the amount of these proteins clearly declined in the fractions preceding the peak of neurosecretory granules. Our results suggest that microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of all other nerve terminals and argue against the hypothesis that such vesicles represent an endocytic byproduct of exocytosis of neurosecretory granules.     

Identification of the vasopressin producing and of the oxytocin producing neurons in the hypothalamic magnocellular neurosecretory system of the rat

Immuno-enzyme histochemical investigations showed that, in the magnocellular hypothalamo-hypophysial neurosecretory system of the rat, vasopressin and oxytocin are synthetized in separate neurons. Both the vasopressin neurons and the oxytocin neurons are present in both the supraoptic and the paraventricular nuclei in about the same number. Preferential location of the two kinds of rat neurosecretory neurons is not as obvious as in the bovine hypothalamus. Their perikarya do not show distinct morphological differences. The two kinds of neurosecretory perikarya are the origin of separate vasopressin-containing and oxytocin-containing axons respectively. In the neural lobe, the distribution of the two different types of axons is described.     

Hypothalamo-hypophysial neurosecretory system of the teleost Trichogaster fasciatus (Bloch & Schneider) with a note on it's vascularization

The hypothalamo-hypophysial neurosecretory (HN) complex of Trichogaster fasciatus is described with the help of in situ staining technique and tissue sections. The neurons of the nucleus preopticus (NPO) give rise to fine neurosecretory axons which form the left and right main tracts. The bulk of the tracts come laterally and enter the middle portion of the neurohypophysis. However, some of them spread out and penetrate the rostral neurohypophysis as well. Hypophysial artery contributes to the formation of primary capillary plexus (PCP) which extends from the subterminal region to the extremity of the anterior neurohypophysis. Structurally, the subterminal region and the anterior neurphoyophysis can be compared to the median eminence (ME) of tetrapods as it is differentiated into ependymal, fibrous and reticular layers. Also, these areas have aboundant neurosecretory and silver positive axons suggesting the possibility of direct transmission of neurohormones into the blood.     

Intrinsic membrane properties of magnocellular neurosecretory neurons recorded in vitro

Over the past 15 years, extracellular recordings from the rat supraoptic and paraventricular nuclei have revealed two populations of endocrine neurons that can be antidromically activated from neurosecretory axons in the neurohypophysis. Both the oxytocinergic and vasopressinergic populations of magnocellular neuroendocrine cells (MNCs) fire bursts of action potentials that facilitate hormone release from neurohypophyseal terminals. Moreover, both populations are osmosensitive, increasing their firing rate as osmolarity is elevated. Recently, slice and explant preparations of hypothalamus have enabled intracellular recording of these cells in normal and modified saline solutions. Spiking, bursting, and osmosensitivity can occur independently of synaptic input, enabling MNCs, for example, to transform an unpatterned depolarizing influence into the repetitive bursting pattern associated with vasopressin release. Current-clamp studies have started to characterize the repertoire of conductances across the MNC membrane that are responsible for action potential discharge, afterpotentials, bursting, and osmosensitivity. This provides a basis not only for further voltage-clamp studies, but for understanding transmitter effects that act by modulating intrinsic MNC currents.     

Ecologic-histophysiological analysis of the neurohypophysis-metaadenohypophysis complex of the sazan female before and after spawning]

Much neurosecretory material (4.4 +/- 0.1 units) is seen in the neurohypophysis of sazan females before spawning. A1, A2, and B type terminals contain numerous neurosecretory granules, in particular, elementary granules. Synaptic vesicles are not numerous. The amount of neurosecretory material decreases during and shortly after spawning down to 3.6 +/- 0.3 units. Neurosecretory terminals have less elementary granules. The number of granulated, disintegrating, residual granules and synaptic vesicles somewhat increases. It is hypothesized that mainly peptide neurohormones are discharged from A1 and A2 terminals in the common circulation, and that both peptide neurohormones and monoamines reach glandular cells of the metaadenohypophysis in sazan females during and immediately spawning. The role played by neurohormones and monoamines in controlling the function of both the metaadenohypophysis glandular cells and visceral organs involved in the general adaptive reactions of the fish organism is discussed.