Vasopressin-secreting Neurons Research Articles

Simplified drug efficacy evaluation system for vasopressin neurodegenerative disease using mouse disease-specific induced pluripotent stem cells

Familial neurohypophyseal diabetes insipidus (FNDI) is a degenerative disorder in which vasopressin-secreting neurons degenerate over time due to the production of mutant proteins. We have demonstrated therapeutic effects of chemical chaperones in an FNDI mouse model, but the complexity and length of this evaluation were problematic. In this study, we established disease-specific mouse induced pluripotent stem cells (iPSCs) from FNDI-model mice and differentiated vasopressin neurons that produced mutant proteins. Fluorescence immunostaining showed that chemical chaperones appeared to protect vasopressin neurons generated from iPSCs derived from FNDI-model mice. Although KCL stimulation released vasopressin hormone from vasopressin neurons generated from FNDI-derived iPSCs, vasopressin hormone levels did not differ significantly between baseline and chaperone-added culture. Semi-quantification of vasopressin carrier protein and mutant protein volumes in vasopressin neurons confirmed that chaperones exerted a therapeutic effect. This research provides fundamental technology for creating in vitro disease models using human iPSCs and can be applied to therapeutic evaluation of various degenerative diseases that produce abnormal proteins.

SAT-427 Low Chloride Transporter Expression in Vasopressin Neurons

Several membrane proteins function as ion pumps/transporters that work to maintain the relative ionic concentrations necessary for neuronal signaling. The potassium-chloride co-transporter two (KCC2) is a symporter that actively pumps K+ and Cl- out of the cell in order to maintain a low intracellular [Cl-], while the sodium-potassium-chloride cotransporters one and two (NKCC1/NKCC2) are symporters that actively pump Na+, K+, and Cl- into the cell. KCC2 and NKCC1/2, therefore, have opposing actions on Cl- transport. In the adult, high KCC2 expression leads to low intracellular [Cl-], which causes GABAA receptor Cl- channels to flux Cl- into the neuron, causing a membrane hyperpolarization. Early in development however, low KCC2 expression reverses the Cl- gradient and causes GABAA receptors to flux Cl- out of the neuron, leading to membrane depolarization and neuronal excitation. The change in the polarity of GABA signaling during development is, in part, due to a shift in KCC2 expression from low to high with the maturation of synaptic circuits. Our previous studies have demonstrated an excitatory GABAA receptor-mediated response due to a GABA equilibrium potential (EGABA) that is shifted positive, like that seen early in development, in adult vasopressin (VP)-secreting neurons of the rat hypothalamus. In the current study, we used immunohistochemistry to compare expression levels of the main Cl- transporters, KCC2, NKCC1, and NKCC2, between VP- and oxytocin (OT)-secreting neurons of the hypothalamic paraventricular and supraoptic nuclei. We found that, in adult male Wistar rats, expression of all three transporters is uniformly lower in VP neurons than in adjacent OT neurons in the hypothalamus. Because the phosphorylation state of KCC2 affects membrane trafficking and stabilization, we are also using Western blot to analyze phosphorylation of KCC2 under different osmotic conditions. Finally, we are using Cre-dependent viral transduction to express a light-activatable Cl- channel conditionally in vasopressin-secreting neurons to test for the effect of Cl- flux on vasopressin secretion.

Vasopressin-secreting neurons derived from human embryonic stem cells through specific induction of dorsal hypothalamic progenitors

Arginine-vasopressin (AVP) neurons exist in the hypothalamus, a major region of the diencephalon, and play an essential role in water balance. Here, we established the differentiation method for AVP-secreting neurons from human embryonic stem cells (hESCs) by recapitulating in vitro the in vivo embryonic developmental processes of AVP neurons. At first, the differentiation efficiency was improved. That was achieved through the optimization of the culture condition for obtaining dorsal hypothalamic progenitors. Secondly, the induced AVP neurons were identified by immunohistochemistry and these neurons secreted AVP after potassium chloride stimulation. Additionally, other hypothalamic neuropeptides were also detected, such as oxytocin, corticotropin-releasing hormone, thyrotropin-releasing hormone, pro-opiomelanocortin, agouti-related peptide, orexin, and melanin-concentrating hormone. This is the first report describing the generation of secretory AVP neurons derived from hESCs. This method will be applicable to research using disease models and, potentially, for regenerative medicine of the hypothalamus.

Bidirectional Anticipation of Future Osmotic Challenges by Vasopressin Neurons

Ingestion of water and food are major hypo- and hyperosmotic challenges. To protect the body from osmotic stress, posterior pituitary-projecting, vasopressin-secreting neurons (VPpp neurons) counter osmotic perturbations by altering their release of vasopressin, which controls renal water excretion. Vasopressin levels begin to fall within minutes of water consumption, even prior to changes in blood osmolality. To ascertain the precise temporal dynamics by which water or food ingestion affect VPpp neuron activity, we directly recorded the spiking and calcium activity of genetically defined VPpp neurons. In states of elevated osmolality, water availability rapidly decreased VPpp neuron activity within seconds, beginning prior to water ingestion, upon presentation of water-predicting cues. In contrast, food availability following food restriction rapidly increased VPpp neuron activity within seconds, but only following feeding onset. These rapid and distinct changes in activity during drinking and feeding suggest diverse neural mechanisms underlying anticipatory regulation of VPpp neurons.

Hold the Salt: Vasopressor Role for BDNF

In a recent publication in Neuron, Choe et al. (2015) demonstrate that brain-derived neurotrophic factor (BDNF) signaling mediates salt-induced blood pressure elevation by increasing the excitability of hypothalamic vasopressin-secreting neurons. These findings suggest complex roles for BDNF in adaptive cardiovascular responses to physiological challenges and in the pathogenesis of hypertension.

Modifications of atrial natriuretic peptide and vasopressin peptides in the rat hypothalamic supraoptic nucleus during resistance training.

Many studies have demonstrated the involvement of atrial natriuretic peptide (ANP) and vasopressin (VP) in the homeostasis of body fluids, but few studies have regarded the hypothalamic magnocellular neurosecretory system during physical exercises. The aim of the present immunohistochemical work is to study the activity of ANP and VP secreting neurons of the hypothalamic supraoptic nucleus during and after resistance training. The study was carried out in Wistar rats trained by a physical resistance-type exercise, using a rung ladder and a varying load fastened to the tail of each rat; the exercise lasted 20 min everyday for periods of 15, 30 and 45 days. Animal groups were sacrificed at the end of each training period and one group was sacrificed after 60 days from the beginning of training, i.e.15 days after completing a 45 day training. The results show that ANP and VP-immunopositivity is at first lesser in the trained rats than in the corresponding controls and then increases from the 15th to the 45th day of training; the increase of the immunopositivity in the trained rats indicates a decreased degranulation of the neurons. The comparison between VP and ANP-immunopositivity suggests that in the early phase of training VP-release in the bloodstream is higher than ANP-release, therefore the antidiuretic action of VP is expected to prevail on ANP action and an electrolyte unbalance may occur.

CANNABINOID-MEDIATED REGULATION OF THE HYPOTHALAMO-PITUITARY-ADRENAL AXIS in rats: AGE DEPENDENT ROLE OF VASOPRESSIN

Adaptation to stress is a fundamental component of life and the hypothalamo-pituitary-adrenocortical axis (HPA) plays a crucial role in it. The place of cannabinoid influence seems to be in the brain, especially where corticotropin releasing hormone and vasopressin (AVP) secreting neurons are located. The role of AVP is considered to be more important in young than in adult rats. Here we addressed the question if cannabinoid-mediated regulation of the HPA involves AVP and if there is any difference between young and adult rats in this process. 10-day-old and adult AVP deficient Brattleboro rats were compared with their heterozygous littermates 1h after WIN 55,212-2 (6mg/kg i.p.) injection. In control animals the injection led to elevated adrenocorticotropin (ACTH) and corticosterone hormone levels at both ages without remarkable age difference in ACTH levels while all corticosterone levels of adults was approximately 10-times higher. The ACTH secretion of young AVP deficient rats failed to react to WIN 55,212-2 injection while their corticosterone levels were even higher than their littermates. In contrast in adult the role of AVP was diminished. We can conclude that the peripheral administration of cannabinoids leads to HPA axis stimulation, which process involves AVP at least in the young rats. The discrepancy between ACTH and corticosterone levels in young rats suggests an alternative adrenal gland regulatory pathway, which might be present in all studied animals. However, it comes to the front just in AVP deficient pups.

Novel Aspects of Glutamatergic Signalling in the Neuroendocrine System

L-glutamate, the main excitatory neurotransmitter, influences virtually all neurones of the neuroendocrine hypothalamus via synaptic mechanisms. Vesicular glutamate transporters (VGLUT1-3), which selectively accumulate L-glutamate into synaptic vesicles, provide markers with which to visualise glutamatergic neurones in histological preparations; excitatory neurones in the endocrine hypothalamus synthesise the VGLUT2 isoform. Results of recent dual-label in situ hybridisation studies indicate that glutamatergic neurones in the preoptic area and the hypothalamic paraventricular, supraoptic and periventricular nuclei include parvocellular and magnocellular neurosecretory neurones which secrete peptide neurohormones into the bloodstream to regulate endocrine functions. Neurosecretory terminals of GnRH, TRH, CRF-, somatostatin-, oxytocin- and vasopressin-secreting neurones contain VGLUT2 immunoreactivity, suggesting the co-release of glutamate with hypophysiotrophic peptides. The presence of VGLUT2 also indicates glutamate secretion from non-neuronal endocrine cells, including gonadotrophs and thyrotrophs of the anterior pituitary. Results of in vitro studies show that ionotropic glutamate receptor analogues can elicit hormone secretion at neuroendocrine/endocrine release sites. Structural constituents of the median eminence, adenohypophysis and neurohypophysis contain elements of glutamatergic transmission, including glutamate receptors and enzymes of the glutamate/glutamine cycle. The synthesis of VGLUT2 exhibits robust up-regulation in response to certain endocrine challenges, indicating that altered glutamatergic signalling may represent an important adaptive mechanism. This review article discusses the newly emerged non-synaptic role of glutamate in neuroendocrine and endocrine communication.

Contribution of TRPV channels to osmosensory transduction, thirst, and vasopressin release

Systemic osmoregulation is an integrated physiological process through which water intake and excretion are continuously balanced against salt intake and excretion to maintain the osmolality of the extracellular fluid near an optimal 'set-point' value. The behaviors (that is, thirst and sodium appetite) and renal responses (diuresis and natriuresis) that are modulated to mediate osmoregulatory homeostasis are mainly controlled by the nervous system. Appropriate regulation of these parameters depends in large part on specialized osmosensitive neurons, termed osmoreceptors, which convert changes in plasma osmolality into electrical signals that ultimately modulate effector functions to achieve homeostasis. Previous work has shown that mechanosensitive cation channels expressed in osmoreceptor neurons play a key role in the process of osmosensory transduction. Although the molecular identity of these channels remains unknown, a growing body of evidence, reviewed here, indicates that members of the transient receptor potential vanilloid family of ion channels may contribute to osmosensory transduction and to homeostatic responses implicated in the control of water balance.

Risk of diabetes insipidus after pituitary surgery

Diabetes insipidus is a common complication of pituitary surgery, occurring acutely in 18–30% of operations. Most patients recover full posterior function but a minority sustain sufficient damage to vasopressin-secreting neurons to develop permanent diabetes insipidus. The triplephase response, comprising early transient diabetes insipidus, followed by a period of antidiuresis and hyponatremia, before resolving into permanent diabetes insipidus, is an uncommon but important variant. Diabetes insipidus is more common following surgery for craniopharyngioma compared with pituitary adenoma, and craniopharyngioma patients may develop the dangerous complication of adipsic diabetes insipidus. Diabetes insipidus is also more common following pituitary surgery for Rathke’s cleft cysts and possible adrenocorticotropic hormone-secreting adenomas. The data on tumor size are conflicting but our own experience would suggest that diabetes insipidus is more common after surgery for large suprasellar tumors, particularly those with hypothalamic extension. The experience and skill of the neurosurgeon also determines the likelihood of developing diabetes insipidus. The diagnosis of postsurgical diabetes insipidus depends on excluding other forms of polyuria, including diabetes mellitus, excess intravenous fluids and therapy with diuretics or mannitol. Treatment with vasopressin analogues is almost always effective in controlling renal water excretion and normalizing plasma sodium concentrations.

Burst initiation and termination in phasic vasopressin cells of the rat supraoptic nucleus: a combined mathematical, electrical, and calcium fluorescence study.

Vasopressin secreting neurons of the rat hypothalamus discharge lengthy, repeating bursts of action potentials in response to physiological stress. Although many electrical currents and calcium-dependent processes have been isolated and analyzed in these cells, their interactions are less well fathomed. In particular, the mechanism of how each burst is triggered, sustained, and terminated is poorly understood. We present a mathematical model for the bursting mechanism, and we support our model with new simultaneous electrical recording and calcium imaging data. We show that bursts can be initiated by spike-dependent calcium influx, and we propose that the resulting elevation of bulk calcium inhibits a persistent potassium current. This inhibition depolarizes the cell above threshold and so triggers regenerative spiking and further calcium influx. We present imaging data to show that bulk calcium reaches a plateau within the first few seconds of the burst, and our model indicates that this plateau occurs when calcium influx is balanced by efflux and uptake into stores. We conjecture that the burst is terminated by a slow, progressive desensitization to calcium of the potassium leak current. Finally, we propose that the opioid dynorphin, which is known to be secreted from the somatodendritic region and has been shown previously to regulate burst length and phasic activity in these cells, is the autocrine messenger for this desensitization.

Single-unit activity of paraventricular nucleus neurons in response to intero- and exteroceptive stressors in conscious, freely moving rats

Extracellular recordings of 114 neurons in the hypothalamic paraventricular nucleus (PVN) of conscious, freely moving male rats were performed using a movable electrode system. Single-unit activities were examined for their spontaneous firing patterns and responses to intero- and exteroceptive stressors, including disturbance in arterial blood pressure, water deprivation, air-jet stimulation, and systemic administration of cholecystokinin-8 (CCK). PVN neurons were assigned to one of two groups on the basis of their spontaneous firing patterns: phasic ( n=29) and non-phasic ( n=85). Intravenous (i.v.) administration of phenylephrine (8 μg/kg) resulted in the inhibition of a greater percentage of phasic-type (88.9%; 24/27) than non-phasic-type neurons (14.9%; 11/74). Most phasic-type neurons showed excitation in response to i.v. administration of sodium nitroprusside (20 μg/kg, 66.7%; 18/27) and water deprivation (15 h, 77.8%; 7/9) when compared to non-phasic-type neurons. Conversely, a greater number of non-phasic-type neurons showed excitation in response to air-jet stimulation (5 l/min, 10 s, 29.0%; 20/69) and to i.v. administration of CCK (5 μg/kg, 24.5%; 11/45) when compared to phasic-type neurons. However, most non-phasic-type neurons that demonstrated excitation in response to i.v. administration of CCK (88.9%; 8/9) did not respond to air-jet stimulation. The present study indicated that phasically firing neurons recorded from the PVN in conscious, freely moving rats are putative vasopressin-secreting neurons on the basis of their responses to intero- and exteroceptive stressors. These data contribute to our understanding of local neural mechanisms within the PVN that are responsible for stress responses in conscious rats.

Lesion of the perinuclear zone attenuates cardiac sensitivity of vasopressinergic supraoptic neurons.

Discrete stretch of the caval-atrial junction decreases the activity of vasopressin-secreting neurons in the supraoptic nucleus (SON). The perinuclear zone (PNZ) of the SON is necessary for inhibition of vasopressin neurons following an increase in blood pressure. To determine whether the PNZ is necessary for cardiopulmonary regulation of vasopressin neurons, male rats received three unilateral injections of the excitotoxin ibotenic acid (n = 9) or phosphate-buffered saline vehicle (n = 10) into the PNZ. Extracellular activity of antidromically identified phasic vasopressin neurons in the ipsilateral SON was recorded. Of the 26 neurons recorded from vehicle-injected rats 26 were inhibited by an increase in blood pressure and 22 of those neurons were sensitive to caval-atrial distension. Of the neurons recorded from PNZ-lesion rats, only 12 of 29 were inhibited by an increase in blood pressure (P < 0.05), and only 11 neurons were sensitive to caval-atrial stretch (P < 0.05). Functional lesion of the PNZ significantly attenuates both arterial and cardiopulmonary baroreceptor-mediated inhibition of supraoptic vasopressin neurons, suggesting that the PNZ is a necessary component of both pathways.

Role of Midbrain Parabrachial Nucleus in Controlling Electrical Activity of Oxytocin and Vasopressin Secreting Neurones in the Hypothalamic Supraoptic Nucleus.

Release of posterior pituitary hormones is known to be controlled by hemodynamic information in addition to factors related to reproductive function such as suckling stimulus by pups during lactation and afferents from the uterine cervix during parturition. The midbrain parabrachial nucleus, which is one of the major sites for the integration of the autonomic function, sends heavy projections to the hypothalamic supraoptic and paraventricular nuclei which contain oxytocin and vasopressin secreting neurones. The present experiments were undertaken to examine the physiological role of these projections. Electrical stimulation of the parabrachial nucleus increased the excitability of 65% of putative oxytocin neurones, and 60% of putative vasopressin neurones. Electrolytic lesion of the parabrachial nucleus itself did not affect the basal firing rate, which suggests that inputs from the parabrachial nucleus to the supraoptic nucleus are not tonically driven. Although the lesion did not change the responsiveness of neurones to hyperosmotic stimulation, it affected the responsiveness of putative vasopressin neurones to the decrease of blood pressure. In addition to the well-known role of neurosecretory neurones in reproductive functions, these neurones may change their electrical activity in response to inputs driven by the parabrachial nucleus and contribute to the control of body fluid balance.

Projections of medullary and pontine noradrenergic neurons to the horizontal limb of the nucleus of diagonal band in the rat

Recent investigations in the rat have implicated a noradrenergic innervation to the horizontal nucleus of the diagonal band of Broca as a critical link in a neural circuit that conveys baroreceptor information centrally to inhibit the firing of vasopressin-secreting neurons in the hypothalamic supraoptic nucleus. In this study we used small intra-diagonal band injections of a retrograde tracer, rhodamine latex microspheres, in combination with tyrosine hydroxylase histochemistry to identify brainstem noradrenergic cells contributing to this innervation. In three cases where tracer injections were limited to the horizontal limb of the diagonal band, we observed 20–50 double-labelled neurons ipsilaterally in the dorsal part of the locus coeruleus (A6), and the caudal nucleus tractus solitarius (A2), and bilaterally in the caudal ventrolateral medulla (A1). Double-labelled neurons were also noted in the ventral tegmental area (dopaminergic A10 cell group). Although all major brainstem noradrenergic cell groups contribute fibers to the horizontal limb of the nucleus of diagonal band, data from physiological studies suggest that the noradrenergic A2 neurons in the nucleus tractus solitarius are the most likely pathway through which it receives this baroreceptor information.

Chapter 2.1.3 Phenotypic and state-dependent expression of the electrical and morphological properties of oxytocin and vasopressin neurones

Oxytocin and vasopressin secreting neurones of the hypothalamic supraoptic nucleus share many membrane characteristics and a roughly similar morphology. However, these two neurone types differ in the relative expression of some intrinsic and synaptic currents, and in the extent of their respective dendritic arbors. Spike depolarizing afterpotentials are present in both types, but more frequently give rise to prolonged burst discharges in vasopressin neurones. Oxytocin, but not vasopressin neurones, are characterized by a depolarization-activated, sustained outward rectifier which turns on near spike threshold, and which can produce prolonged spike frequency adaptation. When this sustained current is deactivated by small hyperpolarizing pulses, a rebound depolarization sufficient to evoke short spike trains follows the offset of these pulses. Both oxytocin and vasopressin neurones exhibit a transient outward rectification underlain by an Ia-type current. This transient rectifier delays spiking to depolarizing stimuli from a relatively hyperpolarized baseline, and is more prominent in vasopressin neurones. As a result, oxytocin neurones may be more reactive to depolarizing inputs. Both cell types receive glutamatergic, excitatory synaptic inputs and both possess R,S- alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptor subtypes. The AMPA receptor channel on both cell types is characterized by a relatively high calcium permeability and voltage-dependent rectification, characteristic of a diminished presence of the GluR2 AMPA subunit. However, AMPA-mediated synaptic transients are larger, and decay faster, in oxytocin compared with vasopressin neurones, suggesting a potential difference for synaptic integration. The characteristics of NMDA-mediated synaptic transients are similar in oxytocin and vasopressin neurones, but some data suggest NMDA receptors may be less involved in the glutamatergic activation of oxytocin neurones. In both cell types, synaptic release of glutamate often coactivates AMPA and NMDA receptors. The dendritic morphology of oxytocin and vasopressin neurones in female rats differs from one another and exhibits considerable plasticity as a function of endocrine state. In virgin rats, oxytocin neurones have more dendritic branches and a greater total dendritic length compared with lactation, when the arbor is much less extensive. A complementary change occurs in vasopressin dendrites, which are more extensive during lactation. This reorganization suggests that oxytocin neurones may be more electronically compact during lactation. In addition, such dramatic shifts in overall dendritic length imply that significant gains and losses in either the total number of synapses, or in synaptic density, are incurred by both cell types as a function of reproductive state.

Opposing effects of intracerebroventricularly injected norepinephrine on oxytocin and vasopressin neurons in the paraventricular nucleus of the rat

Our previous study shows that intracerebroventricularly (i.c.v.) injected norepinephrine (NE) had different effects on the discharge of different firing patterns of magnocellular neurons in the paraventricular nucleus (PVN). In the present study we further classified antidromically identified magnocellular neurons into two groups: vasopressin (VP) and oxytocin (OT) secreting neurons, and found that of all 48 cases of magnocellular neurons, NE had mainly excitatory effects on 36 cases of putatively OT-secreting neurons, and inhibitory effects on 12 cases of VP-secreting neurons. The third ventricular injected NE had almost the same effect on two types of neurons as that of lateral ventricular injection, partly ruling out the possibility that the lateral ventricularly injected NE may have acted indirectly on the magnocellular neurons in PVN. The results show that different mechanisms may be involved in mediating the effect of i.c.v. injected NE on VP- and OT-secreting neurons in the PVN.

Effects of Sinoaortic Denervation on Fos Expression in the Brain Evoked by Hypertension and Hypotension in Conscious Rabbits

We have previously shown [Li and Dampney (1994) Neuroscience 61, 613–634] that periods of sustained hypertension and hypotension each induces a distinctive and reproducible pattern of neuronal expression of Fos (a marker of neuronal activation) in specific regions of the brainstem and forebrain of conscious rabbits. The aim of this study was to determine the contribution of afferent inputs from arterial baroreceptors to the activation of neurons in these various brain regions that is caused by a sustained change in arterial pressure. Experiments were carried out on rabbits in which the carotid sinus and aortic depressor nerves were cut in a preliminary operation. Following a recovery period of seven to 10 days, a moderate hypertension or hypotension (increase or decrease in arterial pressure of 20–30 mmHg) was induced in conscious barodenervated rabbits for 60 min by the continuous infusion of phenylephrine or sodium nitroprusside, respectively. In control experiments, barodenervated rabbits were subjected to the identical procedures except that they were infused with the vehicle solution alone. Compared with the effects seen in barointact rabbits, [Li and Dampney (1994) Neuroscience 61, 613–634] the number of neurons that expressed Fos in response to hypertension was reduced by approximately 90% in the nucleus of the solitary tract and in the caudal and intermediate parts of the ventrolateral medulla. In supramedullary regions, baroreceptor denervation resulted in a reduction of approximately 60% in hypertension-induced Fos expression in the central nucleus of the amygdala and in the bed nucleus of the stria terminalis, but no significant reduction in the parabrachial complex in the pons. Following hypotension, the number of neurons that expressed Fos in barodenervated rabbits, compared with barointact rabbits, [Li and Dampney (1994) Neuroscience 61, 613–634] was reduced by approximately 90% in the nucleus of the solitary tract, area postrema, and caudal, intermediate and rostral parts of the ventrolateral medulla. Baroreceptor denervation also resulted in a similar large reduction in hypotension-induced Fos expression in many supramedullary regions (locus coeruleus, midbrain periaqueductal grey, hypothalamic paraventricular nucleus, and in the central nucleus of the amygdala and the bed nucleus of the stria terminalis in the basal forebrain). In the supraoptic nucleus, hypotension-induced Fos expression in barodenervated rabbits was reduced by 75% compared to barointact animals, but was still significantly greater than in control animals. There was also a high level of Fos expression, much greater than in control animals, in the circumventricular organs surrounding the third ventricle (subfornical organ and organum vasculosum lamina terminalis). The results indicate that in conscious rabbits the activation of neurons that occurs in several discrete regions at all levels of the brain following a sustained change in arterial pressure is largely dependent upon inputs from arterial baroreceptors, with the exception of neurons in the circumventricular organs surrounding the third ventricle that are activated by sustained hypotension. The latter group of neurons are known to project to vasopressin-secreting neurons in the supraoptic nucleus, and may therefore via this pathway trigger the hypotension-induced release of vasopressin that occurs in the absence of baroreceptor inputs.

The glutamatergic innervation of oxytocin- and vasopressin-secreting neurons in the rat supraoptic nucleus and its contribution to lactation-induced synaptic plasticity.

The present ultrastructural study analysed the distribution of glutamatergic synapses on oxytocin- and vasopressin-secreting neurons in the rat supraoptic nucleus (SON) after post-embedding immunogold labelling for glutamate immunoreactivity, visible over synaptic-like vesicles, mitochondria and synaptic densities. Double labelling for glutamate and GABA showed that putative glutamatergic terminals were distinct from GABAergic terminals. In ultrathin sections stained for glutamate and either oxytocin or vasopressin, the proportion of glutamatergic synapses was similar on oxytocinergic and vasopressinergic somata in virgin rats under basal conditions of peptide release as well as in lactating rats, in which oxytocin secretion is enhanced. Cross-sectional soma areas were significantly increased in lactating rats: oxytocinergic profiles were, on average, approximately 40% larger than in virgin rats. However, the incidence of axo-somatic glutamatergic synapses (assessed as mean number of synapses per 100 microm of plasmalemma or proportion of somatic surface apposed to synaptic active zones) did not diminish, indicating that there was a compensatory increase of synapses during lactation. Also, we found an increase in the number of glutamatergic terminals making synaptic contact simultaneously onto two or more oxytocinergic elements in the same plane of section. Our observations therefore indicate that SON oxytocinergic and vasopressinergic neurons are innervated to a similar extent by a relatively large proportion of glutamatergic synapses. They reveal, moreover, that glutamatergic afferents participate in the lactation-induced synaptic plasticity of the oxytocinergic system.

CNS pathways mediating cardiovascular regulation of vasopressin.

1. The release of vasopressin from the neurohypophysial terminals of hypothalamic magnocellular neurosecretory neurons is subject to regulation by peripheral baroreceptors, cardiopulmonary volume receptors and circulating angiotensin II. Information from these sources is transmitted through different pathways to achieve different influences on the excitability of the vasopressin-secreting cells. 2. A brief increase in arterial pressure, sufficient to activate baroreceptors, is associated with a transient and selective GABAergic inhibition of these neurosecretory neurons, achieved through a multisynaptic pathway that involves ascending catecholaminergic fibres and neurons in the diagonal band of Broca. A decrease in arterial pressure activates peripheral low volume receptors and initiating neural inputs that result in an increase in the excitability of vasopressin-secreting neurons, achieved via pathways that include direct projections from caudal ventrolateral medulla A1 neurons. 3. Hypotension also releases renal renin and leads to the formation of angiotensin II; binding of this hormone to AT1 receptors on subfornical organ neurons promotes activation of a central angiotensinergic input that evokes a predominantly excitatory effect on vasopressin neurons.