Neurosecretory Cells Of Supraoptic Nucleus Research Articles

Nav1.9 expression in magnocellular neurosecretory cells of supraoptic nucleus

Osmoregulation in mammals is tightly controlled by the release of vasopressin and oxytocin from magnocellular neurosecretory cells (MSC) of the supraoptic nucleus (SON). The release of vasopressin and oxytocin in the neurohypophysis by axons of MSC is regulated by bursting activity of these neurons, which is influenced by multiple sources, including intrinsic membrane properties, paracrine contributions of glial cells, and extrinsic synaptic inputs. Previous work has shown that bursting activity of MSC is tetrodotoxin (TTX)-sensitive, and that TTX-S sodium channels Nav1.2, Nav1.6 and Nav1.7 are expressed by MSC and upregulated in response to osmotic challenge in rats. The TTX-resistant sodium channels, NaV1.8 and Nav1.9, are preferentially expressed, at relatively high levels, in peripheral neurons, where their properties are linked to repetitive firing and subthreshold electrogenesis, respectively, and are often referred to as “peripheral” sodium channels. Both sodium channels have been implicated in pain pathways, and are under study as potential therapeutic targets for pain medications which might be expected to have minimal CNS side effects. We show here, however, that Nav1.9 is expressed by vasopressin- and oxytocin-producing MSC of the rat supraoptic nucleus (SON). We also show that cultured MSC exhibit sodium currents that have characteristics of Nav1.9 channels. In contrast, Nav1.8 is not detectable in the SON. These results suggest that Nav1.9 may contribute to the firing pattern of MSC of the SON, and that careful assessment of hypothalamic function be performed as NaV1.9 blocking agents are studied as potential pain therapies.

MORPHOFUNCTIONAL CHANGES OF PITUITARY-NEUROHYPOPHYSIS SYSTEM IN IMMATURE RATS WITH STREPTOZOTOCIN DIABETES MELLITUS

The research is devoted to the study of morphofunctional organization of the pituitary-neurohypophysis system of diabetes type 1. It was found that on the 14 th day after the streptozotocin diabetes simulation in 1-month old animals with hyperglycemia the increasing of functional activity of neurosecretory cells of supraoptic nuclei and increasing of bulk density in their neurosecretory granules are observed. Because of such conditions in the nervous lobe of neurohypophysis increase of bulk density of neurosecretory granules by the diffusing and the residual types are observed. On the 42 th day of the experiment constant high hyperglycemia leads to the stage of depletion of neurosecretory cells of supraoptic and paraventricular nuclei and development of dystrophic-destructive changes in them. In the neurohypophysis the reducing of bulk density of young and mature neurosecretory granules is observed. In the pituicytes of type-2 a large number of different sizes and shapes of lipid drops are observed.

Morphofunctional Analysis of Effect of Short-Term Sleep Deprivation on the Wakefulness–Sleep Cycle in Young and Adult Rats

The work presents comparative data on changes of neurophysiological, time characteristics of the wakefulness–sleep cycle (WSC) and morphofunctional state of neurosecretory cells in supraoptic nucleus of the hypothalamus, which develop under influence of a 6-h long sleep deprivation in adult and one-month old rats. It is shown that the rebound of sleep develops in adult animals with a delay, after the 3rd hour and is characterized by a moderate increase of portions of slow-wave (SSP) and fast-wave (FSP) sleep phases in WSC and by a decrease of the wakefulness portion. Morphological analysis of the hypothalamus nonapeptidergic system has revealed a rise of content of neurosecretory material in fibers of supraoptic nucleus cells an in area of supraoptic-pituitary tract, as well as marked hyperemia that indicates activation of processes of secretion of neurohormones into the general blood flow; these reactions are similar to reactions of this system to stress. In rat pups the sleep rebound develops in 0.5 h after the end of the deprivation procedure and is characterized by more pronounced, statistically significant changes in WSC. Individual WSC become very short and almost all of them are completed with episodes of FSP. A statistically significant rise of power of the δ-wave band in electrogram spectra of hippocampus and somatosensory cortex in SSP, whereas peak of the activity in FSP is shifted to α-waves. Ratios of SSP and FSP to wakefulness in individual WSC in mature animals increase after the deprivation 1.53 and 1.85 times, while they are elevated in one-month old animals 5.25 and 6.75 times, respectively. The obtained morphofunctional data allow believing that deprivation is the stress factor of low intensity for adult animals, whereas it may be considered as the stress action of intermediate and even high intensity for rat pups, which changes essentially the interrelations in WSC. Participation of central mechanisms of regulation of sleep and vigilance, which provide processes of compensation of damaging action of deprivation on WSC in the maturing animals, is discussed.

Evidence that multiple P2X purinoceptors are functionally expressed in rat supraoptic neurones.

1. The expression, distribution and function of P2X purinoceptors in the supraoptic nucleus (SON) were investigated by reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and Ca2+-imaging and whole-cell patch-clamp techniques, respectively. 2. RT-PCR analysis of all seven known P2X receptor mRNAs in circular punches of the SON revealed that mRNAs for P2X2, P2X3, P2X4, P2X6 and P2X7 receptors were expressed in the SON, and mRNAs for P2X3, P2X4 and P2X7 were predominant. 3. In situ hybridization histochemistry for P2X3 and P2X4 receptor mRNAs showed that both mRNAs were expressed throughout the SON and in the paraventricular nucleus (PVN). 4. ATP caused an increase in [Ca2+]i in a dose-dependent manner with an ED50 of 1.7 x 10-5 M. The effects of ATP were mimicked by ATPgammaS and 2-methylthio ATP (2MeSATP), but not by AMP, adenosine, UTP or UDP. alphabeta-Methylene ATP (alphabetaMeATP) and ADP caused a small increase in [Ca2+]i in a subset of SON neurones. 5. The P2X7 agonist 2'- & 3'-O-(4-benzoylbenzoyl)-ATP (BzATP) at 10-4 M increased [Ca2+]i, but the potency of BzATP was lower than that of ATP. In contrast, BzATP caused a more prominent [Ca2+]i increase than ATP in non-neuronal cells in the SON. 6. The effects of ATP were abolished by extracellular Ca2+ removal or by the P2 antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), and inhibited by extracellular Na+ replacement or another P2 antagonist, suramin, but were unaffected by the P2X7 antagonist oxidized ATP, and the inhibitor of Ca2+-ATPase in intracellular Ca2+ stores cyclopiazonic acid. 7. Two patterns of desensitization were observed in the [Ca2+]i response to repeated applications of ATP: some neurones showed little or moderate desensitization, while others showed strong desensitization. 8. Whole-cell patch-clamp analysis showed that ATP induced cationic currents with marked inward rectification. The ATP-induced currents exhibited two patterns of desensitization similar to those observed in the [Ca2+]i response. 9. The results suggest that multiple P2X receptors, including P2X3, are functionally expressed in SON neurones, and that activation of these receptors induces cationic currents and Ca2+ entry. Such ionic and Ca2+-signalling mechanisms triggered by ATP may play an important role in the regulation of SON neurosecretory cells.

Ionotropic and metabotropic glutamate receptor agonist-induced [Ca2+]i increase in isolated rat supraoptic neurons.

In the present study, the effects of glutamate and of agonists for ionotropic and metabotropic glutamate receptors on intracellular Ca2+ concentration ([Ca2+]i) were investigated in neurons of the rat supraoptic nucleus (SON). We used the intracellular Ca2+ imaging technique with fura-2, in single magnocellular neurons dissociated from the SON of rats. Glutamate (10(-6)-10(-4) M) evoked a dose-dependent increase in [Ca2+]i. The glutamate agonists exerted similar effects, although with some differences in the characteristics of their responses. The [Ca2+]i response to NMDA was smaller than those of glutamate or the non-NMDA receptor agonists, AMPA and kainate, but was significantly enhanced by the removal of extracellular Mg2+. Glutamate, as well as quisqualate, an agonist for both ionotropic and metabotropic glutamate receptors, evoked a [Ca2+]i increase in a Ca2+-free condition, suggesting Ca2+ release from intracellular Ca2+ stores. This was further evidenced by [Ca2+]i increases in response to a more selective metabotropic glutamate receptor agonist, t-ACPD, in the absence of extracellular Ca2+. Furthermore, the quisqualate-induced Ca2+ release was abolished by the selective metabotropic glutamate receptor antagonist, (S)-4-carboxyphenylglycine. The results suggest that metabotropic glutamate receptors as well as non-NMDA and NMDA receptors are present in the SON neurons, and that activation of the first leads to Ca2+ release from intracellular Ca2+ stores and the activation of the latter two types induces Ca2+ entry. These dual mechanisms of Ca2+ signalling may play a role in the regulation of SON neurosecretory cells by glutamate.

1236 Effects of prenatal stress on the development of corticotropin releasing factor (crf) neurons of hypothalamic paraventricular nucleus (Pvn) in the fetal rat brain

This chapter discusses the influence of muscle receptors on neurons of the supraoptic nucleus (SON). The results presented in the chapter indicate that neurosecretory cells in SON can be excited by electrical and chemical stimulations of muscle receptors and the effect is mediated by Group III and IV fibers without involving large Group I afferents originating from muscle stretch receptors. Respiratory and cardiovascular changes observed during exercise or by stimulations of afferent nerves from the muscle are evoked by impulses in small nerve fibers only and do not seem to involve large Group I afferents although there are some conflicting reports. Kumazawa and Mizumura have shown that some single small diametered fibers in muscle afferents can be excited by mechanical, thermal, and chemical stimuli. It has been suggested that these polymodal fibers may be strongly excited during muscular exercise. The preliminary work shows that gentle mechanical and thermal stimuli applied to the muscle also excite SON neurons, though it is not known whether electrical and chemical stimulations used in the present study also activated nociceptors.

Parabrachial nucleus projection towards the hypothalamic supraoptic nucleus: Electrophysiological and anatomical observations in the rat

It has been proposed that the pontine parabrachial nucleus (PBN) participates in the regulation of body fluid balance and the release of vasopressin from the neurohypophysis, although the pathways mediating the latter response are uncertain. This study in the rat, utilizing anatomical and electrophysiological methods, describes a projection from the lateral PBN towards the hypothalamic supraoptic nucleus (SON). Rats received iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L, 2% solution). After 14-17 days, rats were sacrificed and their brains prepared for immunofluorescent visualization of projections to the SON region. PHA-L-labelled terminals were found primarily in perinuclear regions immediately dorsal to the SON. In contrast, injections within the medial PBN and the nearby Kölliker-Fuse nucleus did not reveal labelling in or around the SON. Extracellular recordings from 86 of 118 antidromically identified neurons in anaesthetized rats revealed a set of complex synaptic responses after stimulation in the PBN. Excitatory responses (in 82 of 86 cells) of short (less than 100 msec, 39/82 cells) and long (greater than 100 msec, 43/82) duration were observed in both vasopressin- and oxytocin-secreting cells of the SON, while 4/86 cells displayed a depressant response to PBN stimulation. In the adjacent perinuclear zone, 22/39 nonneurosecretory cells responded with an increase in their excitability consequent to an identical stimulus. These data suggest a predominantly facilitatory influence of lateral PBN neurons on SON neurosecretory cells in the rat, and that the PBN-SON projection is an indirect one that utilizes an interneuronal network located in the perinuclear zone adjacent to the SON.

Activation of supraoptic neurosecretory cells by local osmotic stimulation of the median preoptic nucleus

In urethane-anesthetized male rats extracellular recordings were obtained from neurosecretory cells in the supraoptic nucleus (SON) while each of electrical stimulation and local osmotic stimulation produced by pressure injection of hypertonic saline was applied to the median preoptic nucleus (MnPO) or to the medial septal nucleus (MS). Electrical stimulation of the MnPO produced orthodromic excitation or initial inhibition followed by strong excitation in most SON cells ( 59 61 ), and local osmotic stimulation of the MnPO excited majority of cells ( 39 51 ). On the contrary, local osmotic stimulation of the MS did not excite SON cells, although electrical stimulation excited all the cells ( 5 5 ). The results suggest that the MnPO is one of the osmosensitive sites controlling electrical activity of SON neurosecretory cells.

Pattern of response in neurosecretory cells of the rat hypothalamic supraoptic nucleus to subiculum stimulation under water deprivation

The effects of water deprivation were investigated in the pattern of response produced by subiculum stimulation in antidromically identified hypothalamic supraoptic neurosecretory cells of lactating rats. In dehydrated animals as compared with the controls, the percentage of neurons responding to subiculum stimulation with an inhibitory action (blockade of antidromic action potential) remained unchanged, although the proportion of differing inhibitory response did alter: numbers of cells with gradually developing inhibitory response increased significantly and fewer cells showed transitory development of inhibition. Inhibitory response emerging as depression of background spike activity showed a quantitative increase, moreover. Plasticity was found to be one distinguishing feature of afferent input from the subiculum to supraoptic nucleus neurosecretory cells and, in particular, a capacity for reorganization under water deprivation.

A1 cell group mediates solitary nucleus excitation of supraoptic vasopressin cells.

Stimulation of the nucleus tractus solitarius (NTS) excites putative vasopressin-secreting cells of the supraoptic nucleus (SON) via a catecholaminergic projection to hypothalamus. Despite recent evidence of a direct catecholaminergic projection from NTS to SON, we have performed single-unit recording experiments in pentobarbital sodium-anesthetized rats to investigate the possibility that NTS stimulation effects on SON vasopressin cells are indirect, being relayed via the A1 noradrenergic cell group of the caudal ventrolateral medulla. The effects of single-pulse NTS and A1 region stimulation on the activity of antidromically identified SON neurosecretory cells that had been functionally characterized as vasopressin secreting were compared. NTS stimulation excited 81% of all putative vasopressin-secreting cells tested (n = 83), with a mean onset latency of 51 +/- 1 ms. A1 region stimulation excited 76% of all cells tested and 90% of units responsive to NTS stimulation, with a mean latency of 39 +/- 1 ms. Consistent with previous work NTS stimulation excited only a minority of oxytocin cells tested (3/13), and of these two-thirds also responded to A1 stimulation. Bilateral electrolytic lesions of the A1 region abolished the effects of NTS stimulation on putative vasopressin cells. Ipsilateral A1 region injections of the inhibitory neurotransmitter gamma-amino-butyric acid reversibly blocked NTS stimulation effects on putative vasopressin cells in animals where the contralateral A1 region had already been lesioned. These results support the proposal that excitation of SON vasopressin-secreting cells after NTS stimulation is due to activation of a relay projection through the A1 noradrenergic cell group of the caudal ventrolateral medulla.

Solitary nucleus excitation of supraoptic vasopressin cells via adrenergic afferents.

Recent studies confirm that the supraoptic nucleus (SON) receives a direct projection from the nucleus of the solitary tract (NTS). We have examined the effect of NTS stimulation on antidromically identified SON neurosecretory cells that were classified as arginine vasopressin (AVP) or oxytocin (OXY) secreting in accord with basal activity patterns and responsiveness to arterial baroreceptor activation. Medial NTS stimulation at the level of the obex or area postrema excited 78% (59 of 76) of putative AVP cells (onset latency 52 +/- 1 ms) but only 20% (3 of 15) of putative OXY cells. Commissural NTS stimulation did not excite AVP cells (n = 13). After complete SON catecholamine afferent disruption, achieved by local injection of 6-hydroxydopamine, AVP cells tested were unresponsive to medial NTS stimulation (12 of 13), although arterial baroreceptor activation inhibited four of four cells. These data indicate that medial NTS stimulation preferentially excites SON AVP cells and that this effect involves an adrenergic input to SON. A direct projection from the A2 catecholamine cell group of the NTS may be involved, although the long latency to excitation and the poor correspondence between effective NTS stimulation sites and the location of the A2 group within NTS raise the possibility that a relay projection, possibly through the A1 catecholamine cell group of the ventrolateral medulla, may be involved.

Lesions of the tissue surrounding the preoptic recess (AV3V region) affect neurosecretory cells in the paraventricular nuclei in the rat

Lesions of the tissue surrounding the preoptic recess (AV3V region) have severe effects on body fluid homeostasis; these include acute adipsia and failure of the antidiuretic response. Because neurosecretory cells in supraoptic nuclei comprise the major source of antidiuretic hormone (ADH) in this species, we have previously observed the fine structure of supraoptic nuclei in rats with AV3V lesions. Paraventricular nuclei are the other major source of ADH in rats. Therefore, in this investigation we compared the fine structure of paraventricular nuclei in rats which had received AV3V lesions 3 days earlier with that of control rats which had received sham lesions and either had drinking water available or had water withheld for 3 days. Degenerating axons and axon terminals were present in paraventricular nuclei of lesioned rats. The degenerating terminals were in axodendritic and less often in axosomatic synapses. Morphometric evaluation revealed that neurosecretory cells did respond to the dehydrated state of the adipsic-lesioned animals, but the response was significantly attenuated compared to that which occurred in sham-lesioned rats deprived of water for 3 days. It appears that AV3V lesions damage afferent connections and impair the response of neurosecretory cells to dehydration in paraventricular as well as supraoptic nuclei. However, in paraventricular nuclei the response is not completely prevented by AV3V lesions during the adipsic period as was observed in supraoptic nuclei. The presence of a response in paraventricular nuclei may be at least partially stimulated by reduced body fluid volume. Information from volume receptors would be carried from the medulla to paraventricular nuclei by ascending pathways which are not affected by AV3V lesions.

Immunocytochemical, Golgi and electron microscopic characterization of putative dendrites in the ventral glial lamina of the rat supraoptic nucleus

Processes of magnocellular neurosecretory cells in the rat supraoptic nucleus which project along the pial surface in the ventral glial lamina were investigated using immunocytochemistry, Golgi stains and electron microscopy. Immunocytochemical studies revealed that although both oxytocin- and vasopressin-containing processes were evident in the ventral glial lamina, vasopressin-containing processes predominated. Ventral processes were thicker and of a different morphology than dorsal axon-like processes which joined the hypothalamo-neurohypophysial tract and exhibited large varicosities along their length or at their apparent termination. Golgi stains revealed that classically defined dendrites of supraoptic neurons projected primarily ventrally and often invaded the ventral glial lamina. No axons were traced to the lamina. Ultrastructurally, processes within the ventral glial lamina characterized as dendrites could be stained immunocytochemically for neurophysin and were post-synaptic to a variety of presynaptic elements. The results suggest that many dendrites from magnocellular neurosecretory cells in the supraoptic nucleus project to the ventral glial lamina and form a restricted, receptive plexus. The previously demonstrated coexistence of catecholamine-containing varicosities and other axon types with these processes in the lamina indicates an important role for supraoptic dendrites in integrating a wide variety of information relevant to neurohypophysial hormone release.

Influence of carotid and aortic baroreceptors on neurosecretory neurons in supraoptic nuclei

(1) The effects of arterial baroreceptor stimulation on the activity of ‘identified’ neurosecretory cells in supraoptic nuclei (SON) of the hypothalamus were investigated in anesthetized cats. (2) Stimulation of baroreceptors by distension of an ‘isolated’ carotid sinus greatly inhibited SON activity. A linear relationship was found to exist between stimulus intensity (an increase in the sinus pressure) and the degree of inhibition of SON neuron activity. This inhibitory effect was abolished by section of the sinus nerve. (3) The inhibitory effect of baroreceptors on SON neuron activity was most pronounced in the first 5 sec during stimulation, and the effect became less when the stimulus lasted for a long period. At the end of stimulation there was a transient reversal of the response. (4) The excitation of aortic baroreceptors by occluding descending aorta strongly inhibited SON neuron activity. (5) Occlusion of carotid arteries augmented the SON neuron activity. Electrical stimulation of the sinus nerve caused an excitation or an inhibition of SON neurons, depending on the stimulus intensities. Apparently these stimuli produced different degrees of excitation in baro- and chemoreceptors. (6) Combination of chemo- and baroreceptor stimulations revealed that the excitatory effect exerted by chemoreceptor stimulation on SON neurons was reversed or blocked by baroreceptor activation, indicating the powerful inhibitory influence of baroreceptors.