Activity Of Magnocellular Neurons Research Articles

Unique Organization of Actin Cytoskeleton in Magnocellular Vasopressin Neurons in Normal Conditions and in Response to Salt-Loading.

Magnocellular neurosecretory cells (MNCs) are intrinsically osmosensitive and can be activated by increases in blood osmolality, triggering the release of antidiuretic hormone vasopressin (VP) to promote water retention. Hence, the activity of magnocellular VP neurons is one of the key elements contributing to the regulation of body fluid homeostasis in healthy organisms. Chronic exposure to high dietary salt leads to excessive activation of VP neurons, thereby elevating levels of circulating VP, which can cause increases in blood pressure contributing to salt-dependent hypertension. However, the molecular basis underlying high-salt diet-induced hyperactivation of magnocellular VP neurons remains not fully understood. Previous studies suggest that magnocellular neurosecretory neurons contain a subcortical layer of actin filaments and pharmacological stabilization of this actin network potentiates osmotically-induced activation of magnocellular neurons. Using super-resolution imaging in situ, we investigated the organization of the actin cytoskeleton in rat MNCs under normal physiological conditions and after a chronic increase in blood osmolality following 7 d of salt-loading (SL). We found that, in addition to the subcortical layer of actin filaments, magnocellular VP neurons are endowed with a unique network of cytoplasmic actin filaments throughout their somata. Moreover, we revealed that the density of both subcortical and cytoplasmic actin networks in magnocellular VP neurons is dramatically increased following SL. These results suggest that increased osmo-responsiveness of VP neurons following chronic exposure to high dietary salt may be mediated by the modulation of unique actin networks in magnocellular VP neurons, possibly contributing to elevated blood pressure in this condition.

PC-FACS

PC-FACS

Vasopressin, Central Autonomic Control and Blood Pressure Regulation

We present recent advances in understanding of the role of vasopressin as a neurotransmitter in autonomic nervous system control of the circulation, emphasizing hypothalamic mechanisms in the paraventricular nucleus (PVN) involved in controlling sympathetic outflow toward the cardiovascular system. Suggest that somato-dendritically released vasopressin modulates the activity of magnocellular neurons in the PVN and SON, their discharge pattern and systemic release. Advances have been made in uncovering autocrine and paracrine mechanisms controlling presympathetic neuron activity, involving intranuclear receptors, co-released neuroactive substances and glia. It is now obvious that intranuclear release of vasopressin and the co-release of neuroactive substances in the PVN, as well as the level of expression of vasopressin receptors, modulate sympathetic outflow to the cardiovascular system and determine vulnerability to stress. Further research involving patho-physiological models is needed to validate these targets and foster the development of more efficient treatment.

Erratum for "Anti-Opioid Effects of RFRP-3 on Magnocellular Neuron Activity in Morphine-Naïve and Morphine-Treated Female Rats?"

Erratum for "Anti-Opioid Effects of RFRP-3 on Magnocellular Neuron Activity in Morphine-Naïve and Morphine-Treated Female Rats?"

Effect of Melanotan‐II on Brain Fos Immunoreactivity and Oxytocin Neuronal Activity and Secretion in Rats

Melanocortins stimulate the central oxytocin systems that are involved in regulating social behaviours. Alterations in central oxytocin have been linked to neurological disorders such as autism, and melanocortins have been proposed for therapeutic treatment. In the present study, we investigated how systemic administration of melanotan-II (MT-II), a melanocortin agonist, affects oxytocin neuronal activity and secretion in rats. The results obtained show that i.v., but not intranasal, administration of MT-II markedly induced Fos expression in magnocellular neurones of the supraoptic (SON) and paraventricular nuclei (PVN) of the hypothalamus, and this response was attenuated by prior i.c.v. administration of the melanocortin antagonist, SHU-9119. Electrophysiological recordings from identified magnocellular neurones of the SON showed that i.v. administration of MT-II increased the firing rate in oxytocin neurones but did not trigger somatodendritic oxytocin release within the SON as measured by microdialysis. Our data suggest that, after i.v., but not intranasal, administration of MT-II, the activity of magnocellular neurones of the SON is increased. Because previous studies showed that SON oxytocin neurones are inhibited in response to direct application of melanocortin agonists, the actions of i.v. MT-II are likely to be mediated at least partly indirectly, possibly by activation of inputs from the caudal brainstem, where MT-II also increased Fos expression.

Magnocellular Neurons and Posterior Pituitary Function.

The posterior pituitary gland secretes oxytocin and vasopressin (the antidiuretic hormone) into the blood system. Oxytocin is required for normal delivery of the young and for delivery of milk to the young during lactation. Vasopressin increases water reabsorption in the kidney to maintain body fluid balance and causes vasoconstriction to increase blood pressure. Oxytocin and vasopressin secretion occurs from the axon terminals of magnocellular neurons whose cell bodies are principally found in the hypothalamic supraoptic nucleus and paraventricular nucleus. The physiological functions of oxytocin and vasopressin depend on their secretion, which is principally determined by the pattern of action potentials initiated at the cell bodies. Appropriate secretion of oxytocin and vasopressin to meet the challenges of changing physiological conditions relies mainly on integration of afferent information on reproductive, osmotic, and cardiovascular status with local regulation of magnocellular neurons by glia as well as intrinsic regulation by the magnocellular neurons themselves. This review focuses on the control of magnocellular neuron activity with a particular emphasis on their regulation by reproductive function, body fluid balance, and cardiovascular status. © 2016 American Physiological Society. Compr Physiol 6:1701-1741, 2016.

Anti-opioid Effects of RFRP-3 on Magnocellular Neuron Activity in Morphine-naïve and Morphine-treated Female Rats.

Neuropeptide FF receptors (NPFFR1 and NPFFR2) have been proposed to possess anti-opioid properties, and be involved in the development of opiate tolerance and dependence. However, there is no evidence to date supporting such opioid effects at the cellular level in vivo. Using in vivo electrophysiological recordings from vasopressin and oxytocin neurons in the supraoptic nucleus, we aimed to determine the effects of NPFFRs on opiate inhibition, tolerance, and dependence at a cellular level. Both vasopressin and oxytocin neurons are acutely inhibited by opioids and develop opiate tolerance. Oxytocin neurons also develop cellular opiate dependence and undergo withdrawal hyperexcitation upon cessation of opiate administration. Here, the classical μ-opioid receptor agonist, morphine robustly inhibited the spontaneous firing rate of vasopressin and oxytocin neurons, and this inhibition was attenuated by pretreatment with the NPFFR1 agonist, RFamide-related peptide-3. In rats infused with morphine for 6 d, vasopressin neurons were unresponsive to morphine, indicating the development of cellular tolerance, but pretreatment with the NPFFR antagonist, GJ14, restored acute morphine inhibition. In morphine-infused rats, RFamide related peptide-3 did not induce withdrawal excitation in oxytocin neurons and GJ14 did not reverse naloxone-precipitated withdrawal excitation. This is the first evidence of anti-opioid effects of the NPFFR system at a cellular level in vivo. Our results suggest that the anti-opioid properties of the NPFFR system reduce morphine sensitivity during tolerance but that it is not involved in dependence.

Effect of interaction between acute administration of morphine and cannabinoid compounds on spontaneous excitatory and inhibitory postsynaptic currents of magnocellular neurons of supraoptic nucleus.

Opioids and cannabinoids are two important compounds that have been shown to influence the activity of magnocellular neurons (MCNs) of supraoptic nucleus (SON). The interaction between opioidergic and cannabinoidergic systems in various structures of the brain and spinal cord is now well established, but not in the MCNs of SON. In this study, whole cell patch clamp recording of neurons in rat brain slice was used to investigate the effect of acute morphine and cannabinoid administration on spontaneous inhibitory and excitatory spostsynaptic currents (sIPSCs and sEPSCs) in MCNs. Bath application of morphine produced an increase in sEPSCs frequency and a decrease in sIPSCs frequency. In contrast, bath application of URB597 (fatty acid amide hydrolase (FAAH) inhibitor) produced a decrease in sEPSCs frequency but an increase in sIPSCs frequency. WIN55212-2 (cannabinoid receptor agonist) decreased both sIPSCs and sEPSCs frequencies of MCNs. Co-application of morphine and URB597 attenuated the effect of morphine on MCNs. Taken together, these data indicated that at the cellular level, pharmacological augmentation of endocannabinoids could attenuate morphine effects on MCNs.

Systemic Leptin Increases the Electrical Activity of Supraoptic Nucleus Oxytocin Neurones in Virgin and Late Pregnant Rats

In the rat hypothalamus, fasting attenuates the expression of oxytocin and this can be reversed by exogenous leptin administration. In the present study, we investigated the effects of systemically administered leptin on the electrical activity of magnocellular neurones in the supraoptic nucleus of urethane-anaesthetised rats. In virgin female rats, systemic leptin significantly excited identified oxytocin neurones with no detected effects on the patterning of activity, as reflected by hazard function analyses. The lowest dose that was consistently effective was 100 μg/i.v., and this dose had no significant effect on vasopressin neurones. In virgin rats fasted overnight, the spontaneous firing rate of oxytocin neurones was significantly lower than in unfasted rats, although leptin had a similar excitatory effect as in unfasted rats. In late pregnant rats (days 19-21 of pregnancy), spontaneous firing rates of oxytocin neurones were higher than in virgins, and the initial response to leptin was similar to that in virgin rats, although the increase in activity was more persistent. In fasted pregnant rats, the mean spontaneous firing rate of oxytocin neurones was again lower than in unfasted rats, although leptin had no significant effect even at the higher dose of 1 mg/rat. Thus, fasting reduced the spontaneous firing rates of oxytocin neurones in nonpregnant rats, and this effect could be reversed by the excitatory effects of leptin. Pregnant rats showed some evidence of leptin resistance but only after an overnight fast.

Differential modulation of N‐type calcium channels by µ‐opioid receptors in oxytocinergic versus vasopressinergic neurohypophysial terminals

Opioids modulate the electrical activity of magnocellular neurons (MCN) and inhibit neuropeptide release at their terminals in the neurohypophysis. We have previously shown that micro-opioid receptor (MOR) activation induces a stronger inhibition of oxytocin (OT) than vasopressin (AVP) release from isolated MCN terminals. This higher sensitivity of OT release is due, at least in part, to the selective targeting of R-type calcium channels. We now describe the underlying basis for AVP's weaker inhibition by MOR activation and provide a more complete explanation of the complicated effects on neuropeptide release. We found that N-type calcium channels in AVP terminals are differentially modulated by MOR; enhanced at lower concentrations but increasingly inhibited at higher concentrations of agonists. On the other hand, N-type calcium channels in OT terminals were always inhibited. The response pattern in co-labeled terminals was analogous to that observed in AVP-containing terminals. Changes in intracellular calcium concentration and neuropeptide release corroborated these results as they showed a similar pattern of enhancement and inhibition in AVP terminals contrasting with solely inhibitory responses in OT terminals to MOR agonists. We established that fast translocation of Ca(2+) channels to the plasma membrane was not mediating current increments and thus, changes in channel kinetic properties are most likely involved. Finally, we reveal a distinct Ca-channel beta-subunit expression between each type of nerve endings that could explain some of the differences in responses to MOR activation. These results help advance our understanding of the complex modulatory mechanisms utilized by MORs in regulating presynaptic neuropeptide release.

Glial Cells: Indispensable Partners of Hypothalamic Magnocellular Neurones

The hypothalamo-neurohypophysial system is comprised of magnocellular neurones that synthesise the neuropeptides oxytocin or vasopressin. As neurohormones, these peptides intervene in the regulation of vital functions such as parturition, lactation, osmotic and cardiovascular regulation. The release of these peptides in the general circulation depends on the electrical activity of their parent neurones, which in turn is regulated by the activity of their afferent inputs conveying distinct information. Thus, in view of the diversity of information governing the activity of magnocellular neurones, it is crucial that the system adapts the appropriate release of oxytocin and vasopressin upon physiological demand. Until recently, it was considered that only neurones could provide such adaptation and regulation. However, a third partner of the synapse, the astrocyte, has been shown to provide further control. Astrocytic processes are in proximity of the magnocellular neurones and their synapses, well positioned to detect and modulate synaptic signals. For instance, astrocytes detect a synaptic signal owing to their diverse neurotransmitter/neuropeptide receptors. In addition, they release a variety of neuroactive substances (i.e. gliotransmitters), which in turn modulate synaptic activity. An important gliotransmitter is the amino acid, d-serine, which, together with glutamate, activates NMDA receptors. Once activated, NMDA receptors govern the weight of individual inputs on magnocellular neurones and thus the impact of distinct types of information on neuronal activity. As reviewed here, numerous observations show that astrocytes must be considered as key elements in the functioning of the hypothalamo-neurohypophysial system.

Localization and osmotic regulation of vesicular glutamate transporter-2 in magnocellular neurons of the rat hypothalamus

In this report we present immunocytochemical and in situ hybridization evidence that magnocellular vasopressin and oxytocin neurons in the hypothalamic supraoptic and paraventricular nuclei express type-2 vesicular glutamate transporter, a marker for their glutamatergic neuronal phenotype. To address the issue of whether an increase in magnocellular neuron activity coincides with the altered synthesis of the endogenous glutamate marker, we have introduced a new dual-label in situ hybridization method which combines fluorescent and autoradiographic signal detection components for vasopressin and vesicular glutamate transporter-2 mRNAs, respectively. Application of this technique provided evidence that 2% sodium chloride in the drinking water for 7 days produced a robust and significant increase of vesicular glutamate transporter-2 mRNA in vasopressin neurons of the supraoptic nucleus. The immunocytochemical labeling of pituitary sections, followed by the densitometric analysis of vesicular glutamate transporter-2 immunoreactivity in the posterior pituitary, revealed a concomitant increase in vesicular glutamate transporter-2 protein levels at the major termination site of the magnocellular axons. These data demonstrate that magnocellular oxytocin as well as vasopressin cells contain the glutamatergic marker vesicular glutamate transporter-2, similarly to most of the parvicellular neurosecretory neurons examined so far. The robust increase in vesicular glutamate transporter-2 mRNA and immunoreactivity after salt loading suggests that the cellular levels of vesicular glutamate transporter-2 in vasopressin neurons are regulated by alterations in water–electrolyte balance. In addition to the known synaptic actions of excitatory amino acids in magnocellular nuclei, the new observations suggest novel mechanisms whereby glutamate of endogenous sources can regulate magnocellular neuronal functions.

Nitric oxide modulation of the hypothalamo-neurohypophyseal system.

Nitric oxide (NO), a free radical gas produced endogenously from the amino acid L-arginine by NO synthase (NOS), has important functions in modulating vasopressin and oxytocin secretion from the hypothalamo-neurohypophyseal system. NO production is stimulated during increased functional activity of magnocellular neurons, in parallel with plastic changes of the supraoptic nucleus (SON) and paraventricular nucleus. Electrophysiological data recorded from the SON of hypothalamic slices indicate that NO inhibits firing of phasic and non-phasic neurons, while L-NAME, an NOS inhibitor, increases their activity. Results from measurement of neurohypophyseal hormones are more variable. Overall, however, it appears that NO, tonically produced in the forebrain, inhibits vasopressin and oxytocin secretion during normovolemic, isosmotic conditions. During osmotic stimulation, dehydration, hypovolemia and hemorrhage, as well as high plasma levels of angiotensin II, NO inhibition of vasopressin neurons is removed, while that of oxytocin neurons is enhanced. This produces a preferential release of vasopressin over oxytocin important for correction of fluid imbalance. During late pregnancy and throughout lactation, fluid homeostasis is altered and expression of NOS in the SON is down- and up-regulated, respectively, in parallel with plastic changes of the magnocellular system. NO inhibition of magnocellular neurons involves GABA and prostaglandin synthesis and the signal-transduction mechanism is independent of the cGMP-pathway. Plasma hormone levels are unaffected by i.c.v. 1H-[1, 2, 4]oxadiazolo-[4,3-a]quinoxalin-1-one (a soluble guanylyl cyclase inhibitor) or 8-Br-cGMP administered to conscious rats. Moreover, cGMP does not increase in homogenates of the neural lobe and in microdialysates of the SON when NO synthesis is enhanced during osmotic stimulation. Among alternative signal-transduction pathways, nitrosylation of target proteins affecting activity of ion channels is considered.

Isoluminant stimuli and red background attenuate the effects of transient spatial attention on temporal resolution

The effects of transient spatial attention on temporal resolution were recently studied and compared with attentional effects on spatial resolution. It was found that in contrast to the attentional enhancement of spatial resolution, transient attention impairs temporal resolution. To account for these findings a novel attentional mechanism was suggested. This attentional mechanism facilitates parvocellular neurons at the attended location, which in turn inhibit the activity of magnocellular neurons at the same location [Yeshurun & Levy, Psychol. Sci. 14 (3) (2003a) 225]. The goal of the present study was to replicate the attentional decrement in temporal resolution (Experiment 1), and perform direct tests of this `parvo facilitation––magno inhibition' hypothesis. The employment of isoluminant stimuli (Experiments 2a and 2b) or a red background (Experiment 3) ensured that the parvo system was the main system mediating performance. Consequently, any parvo–magno inhibitory processes elicited by the attentional mechanism should only have a minor effect on performance. As predicted, these manipulations either significantly attenuated or completely eliminated the attentional decrement in temporal resolution. These findings provide direct support to the hypothesis that attention favors parvocellular over magnocellular neurons.

Activation of postsynaptic GABAB receptors modulate the firing activity of supraoptic oxytocin and vasopressin neurones: role of calcium channels.

Oxytocin and vasopressin release from neurohypophysial terminals is closely related to the firing activity of magnocellular neurones in the supraoptic (SON) and paraventricular nuclei. It is well established that activation of GABAA receptors potently inhibits the activity of SON neurones and, thus, hormone release. However, whether postsynaptic GABAB receptors are expressed in magnocellular neurones, and the role they play in controlling their firing activity, is still controversial. In the present work, we combined immunohistochemical and electrophysiological techniques to determine whether activation of GABAB receptors in identified oxytocin and vasopressin neurones modulates their firing activity. Patch-clamp recordings from SON neurones were obtained either in the slice preparation or from acutely dissociated neurones. Activation of GABAB receptors with the selective agonist baclofen (10 micro m) inhibited voltage-gated Ca2+ currents, reduced the duration of individual action potentials, as well as the magnitude of the hyperpolarizing after-potential. SON firing activity was reduced by baclofen, and effect that was accompanied by a small membrane hyperpolarization. The inhibition of firing discharge persisted in the presence of synaptic blockade media, and was also observed in acutely dissociated SON neurones. Finally, GABAB-mediated modulation of firing activity was largely blocked by the Ca2+ channel blocker Co2+ (2 mm). In general, baclofen modulatory actions were significantly larger, or observed more predominantly, in vasopressin neurones. In summary, these results support the expression of functional postsynaptic GABAB receptors in SON neurones, activation of which efficiently modulates neuronal excitability, in a Ca2+- and cell-type dependent manner.

On the use of red stimuli to isolate magnocellular responses in psychophysical experiments: a perspective.

Neurophysiological recordings have shown that activity of magnocellular neurons may be reduced by red backgrounds. This has led some researchers to use red light, or red filters, in attempts to determine the magnocellular contribution to psychophysical tasks. This requires that red light not affect parvocellular neurons, or at least that it is possible to control for the effect on the parvocellular system by using other colors. The present report investigates these assumptions by calculating the effect of red, green, and blue filters on the three cone pigments and on the four parvocellular color-opponent cell mechanisms. It is found that a red filter has a large effect on the long- and middle-wavelength cone pigments and on the red-green color-opponent mechanisms. A green filter, on the other hand, has little effect. A blue filter has a fairly pronounced effect but this effect is distinctly different from that of the red filter. These results indicate that one ought not rely upon red light to isolate magnocellular activity in psychological experiments. The results also indicate that it is difficult to use colors other than red to control for the effect of this color on the parvocellular system.

Japanese Society for Animal Reproduction: award for outstanding research 2002. Mechanisms controlling neurohypophysial hormone release in the rat.

To explore mechanisms of osmotic control of neurohypophysial hormone release, electrical activity of magnocellular neurons (MCNs) in the hypothalamus, related neurons and efferent renal sympathetic nerve activity (RSNA) were recorded from urethane-anesthetized rats. Local osmotic stimulation applied to organum vasculosum of the lamina terminalis (OVLT) or median preoptic nucleus (MnPO) excited MCNs. Although OVLT neurons projected to MCNs were unresponsive to hyperosmotic stimulation, those projected to MnPO and also receiving excitatory inputs from MCNs, were excited by it. MnPO neurons, which were driven by OVLT stimulation and projected to MCNs, were also osmosensitive. Excitatory connections thus exist from MCNs to OVLT, from OVLT to MnPO and from MnPO to MCNs. Neurons in each of these connections were osmosensitive. This circuit thus appears to constitute an osmoreceptor complex essential for the osmoreception of MCNs. MnPO neurons constituting a part of the osmosensitive circuit were also sensitive to hemodynamic change. Thus this circuit may integrate hemodynamic and osmotic information. Local anesthesia of MnPO diminished activation of RSNA and pressor response induced by third cerebroventricular injection of hypertonic saline. The results suggest that the osmosensitive circuit is involved in body fluid regulation not only by controlling vasopressin secretion but also by modulating sympathetic outflow.

Nicotinic cholinergic activation of magnocellular neurons of the hypothalamic paraventricular nucleus

The aim of the present work was to determine whether paraventricular neurons possess functional acetylcholine nicotinic receptors. Using infrared videomicroscopy and differential interference contrast optics, we performed whole-cell recordings in hypothalamic slices containing the paraventricular nucleus. Acetylcholine, locally applied by pressure microejection in the presence of the muscarinic antagonist atropine, evoked a rapidly rising inward current in paraventricular magnocellular endocrine neurons. This current persisted in the presence of blockers of synaptic transmission. It could be reversibly suppressed by nanomolar concentrations of methyllycaconitine, a selective antagonist of α7-containing nicotinic receptors, but was insensitive to micromolar concentrations of dihydro-β-erythroidine, an antagonist acting preferentially on non-α7 nicotinic receptors. In addition, the effect of acetylcholine could be mimicked by exo-2-(2-pyridyl)-7-azabicyclo[2.2.1]heptane, a recently synthesized nicotinic agonist specific for α7 receptors. Acetylcholine also desensitized paraventricular nicotinic receptors. Desensitization was pronounced and recovery from desensitization was rapid, consistent with the notion that paraventricular nicotinic receptors contain the α7 subunit. Nicotinic currents could not be evoked in paraventricular parvocellular neurons, suggesting that these neurons are devoid of functional nicotinic receptors. The electrophysiological data were corroborated by light microscopic autoradiography, showing that [ 125I]α-bungarotoxin binding sites are present in all the magnocellular divisions of the paraventricular nucleus but are undetectable in other areas of this nucleus. Immunohistochemistry, performed using antibodies directed against vasopressin and oxytocin, indicated that responsiveness to nicotinic agonists was a property of vasopressin as well as of oxytocin magnocellular endocrine neurons, in both the paraventricular and the supraoptic nucleus. We conclude that nicotinic agonists can influence the magnocellular neurosecretory system by directly increasing the excitability of magnocellular neurons. By contrast, they are probably without direct effects on paraventricular parvocellular neurons.

The Vasopressin Receptors Colocalize with Vasopressin in the Magnocellular Neurons of the Rat Supraoptic Nucleus and Are Modulated by Water Balance

Activity of the magnocellular neurons that synthesize vasopressin in the supraoptic and paraventricular nuclei of the hypothalamus is modulated by local release of the neuropeptide within the nuclei. V1a and V1b vasopressin receptor genes are expressed in these cells. The present study reports the localization of V1a and V1b receptors using multiple labeling immunocytochemistry. Both receptors are mainly located in vasopressinergic magnocellular neurons and colocalized with vasopressin in cytoplasmic vesicles dispersed throughout the cell. Possible functional modifications of the mRNA and protein levels of the V1a receptor, the major isoform, were also investigated by semiquantitative in situ hybridization and immunocytochemistry in rats submitted to reduced or increased water intake. V1a mRNA and receptor levels varied with water balance. V1a mRNA level dropped in rats submitted to high water intake. Conversely, dehydration up-regulated the V1a receptor content. These observations suggest that the pathways that regulate the expression of the genes encoding vasopressin and the V1a receptor are linked, which fits the present findings that the two partners are colocalized in cytoplasmic vesicles. Colocalization might explain how V1 autoreceptors are controlled by cell activity and/or local concentration of vasopressin (released locally by the neurons themselves), allowing fine adjustment of magnocellular neuron activity.

The vasopressin receptors colocalize with vasopressin in the magnocellular neurons of the rat supraoptic nucleus and are modulated by water balance.

Activity of the magnocellular neurons that synthesize vasopressin in the supraoptic and paraventricular nuclei of the hypothalamus is modulated by local release of the neuropeptide within the nuclei. V(1a) and V(1b) vasopressin receptor genes are expressed in these cells. The present study reports the localization of V(1a) and V(1b) receptors using multiple labeling immunocytochemistry. Both receptors are mainly located in vasopressinergic magnocellular neurons and colocalized with vasopressin in cytoplasmic vesicles dispersed throughout the cell. Possible functional modifications of the mRNA and protein levels of the V(1a) receptor, the major isoform, were also investigated by semiquantitative in situ hybridization and immunocytochemistry in rats submitted to reduced or increased water intake. V(1a) mRNA and receptor levels varied with water balance. V(1a) mRNA level dropped in rats submitted to high water intake. Conversely, dehydration up-regulated the V(1a) receptor content. These observations suggest that the pathways that regulate the expression of the genes encoding vasopressin and the V(1a) receptor are linked, which fits the present findings that the two partners are colocalized in cytoplasmic vesicles. Colocalization might explain how V(1) autoreceptors are controlled by cell activity and/or local concentration of vasopressin (released locally by the neurons themselves), allowing fine adjustment of magnocellular neuron activity.