Rat Subfornical Organ Research Articles

Drinking attenuates the noradrenaline release in the lateral hypothalamic area induced by angiotensin II activation of the subfornical organ in rats

Experiments were conducted to investigate the role of noradrenergic systems in the lateral hypothalamus area (LHA) in the water intake caused by injection of angiotensin II (ANG II) into the subfornical organ (SFO) in rats. Intracerebral microdialysis techniques were utilized to quantify the extracellular content of noradrenaline (NA) in the LHA. Microinjection of ANG II into the SFO significantly increased NA release in the LHA when water was not available for drinking. The increase in the release of NA in the LHA was significantly attenuated by water intake. In urethane-anesthetized rats, injections of ANG II into the SFO significantly enhanced the release of NA in the LHA that accompanied an elevation in mean arterial pressure (MAP). Intravenous administration of the α-agonist metaraminol, on the other hand, significantly decreased the NA release in the LHA that accompanied an increase in MAP, suggesting that the enhanced NA release in the LHA caused by ANG II into the SFO may be not mediated by increasing in arterial pressure. These results show the involvement of the noradrenergic systems in the LHA in the dipsogenic response induced by angiotensinergic activation of the SFO.

Selective potentiation of N-type calcium channels by angiotensin II in rat subfornical organ neurones.

1. Here we have characterized Ca(2+) currents in rat subfornical organ neurones, and their modulation by angiotensin II. Currents were of the high voltage-activated (HNA) subtype, as the threshold for activation was near -30 mV (mid-point potential (V(50)) of activation -14 mV). Using Ba(2+) as the charge carrier, little inactivation was observed, and it occurred only at depolarized potentials (V(50) of inactivation -12 mV). More inactivation was observed using Ca(2+) as the charge carrier, indicating that Ca(2+)-dependent inactivation plays a role in regulating Ca(2+) channel function in subfornical organ (SFO) neurones. 2. The net Ba(2+) current could be blocked by Cd(2+) (EC(50) 1.6 microM), confirming that currents are of the HVA variety. By using selective antagonists, we identified the presence of both L- and N-type channels; 20 microM nifedipine blocked 22 +/- 1 % of the current, while omega-conotoxin GVIA blocked 65 +/- 7 %, indicating that these currents make up the net current through Ca(2+) channels. 3. Angiotensin II potentiated the inward current throughout the range of activation. Using depolarizing voltage ramps, 1 nM angiotensin potentiated the peak current by 14 +/- 5 %. We then used selective blockade of the HVA component currents; 20 microM nifedipine failed to prevent the potentiation by angiotensin II (12 +/- 5 %), while blocking N-type channels with omega-conotoxin GVIA blocked the facilitation by ANG (2.3 +/- 2 %). Losartan (1 microM) prevented the actions of ANG on the inward current (1.6 +/- 1 %), indicating that the selective effects of ANG on N-type channels in SFO neurones are mediated by AT(1) receptors.

Hemorrhage activates catecholaminergic neurons sensitive to GABA in the nucleus of the solitary tract with ascending projections to the subfornical organ in rats

The activity of neurons in the region of the nucleus of the solitary tract (NTS) that were antidromically identified by electrical stimulation of the rat subfornical organ (SFO) was tested for a response to microiontophoretic application of γ-aminobutyric acid (GABA), hemorrhage (10 ml/kg b.w.t.), or local administration of the chemical neurotoxin, 6-hydroxydopamine (6-OHDA), into the SFO stimulation site. Microiontophoretically (MIPh) applied GABA caused a decrease excitability in 22 out of 24 neurons tested, and the inhibition was blocked by MIPh-applied bicuculline, a GABA A antagonist, but not by phaclofen, a GABA B antagonist. Of these neurons that responded to GABA, 17 displayed an increase in neural firing in response to hemorrhage, while 5 were unresponsive. The occurrence of both antidromic spikes and post-stimulus inhibition of 9 out of 13 neurons tested was completely abolished by the injection of 6-OHDA into the SFO. These results suggest that neurons in the region of the NTS, which carry peripheral baroreceptor information to the SFO, receive GABAergic inhibitory inputs via a GABA A receptor mechanism, and imply that part of these neurons are catecholaminergic.

Muscarinic modulation of GABAergic transmission to neurons in the rat subfornical organ.

Cholinergic actions on subfornical organ (SFO) neurons in rat slice preparations were studied by using whole cell voltage- and current-clamp recordings. In the voltage-clamp recordings, carbachol and muscarine decreased the frequency of GABAergic inhibitory postsynaptic currents (IPSCs) in a dose-dependent manner, with no effect on the amplitudes or the time constants of miniature IPSCs. Meanwhile, carbachol did not influence the amplitude of the outward currents induced by GABA. Furthermore, carbachol and muscarine also elicited inward currents in a TTX-containing solution. From the current-voltage relationship, the reversal potential was estimated to be -7.1 mV. These carbachol-induced responses were antagonized by atropine. In the current-clamp recordings, carbachol depolarized the membrane with increased frequency of action potentials. These observations suggest that acetylcholine suppresses GABA release through muscarinic receptors located on the presynaptic terminals. Acetylcholine also directly affects the postsynaptic membrane through muscarinic receptors, by opening nonselective cation channels. A combination of these presynaptic and postsynaptic actions may enhance activation of SFO neurons by acetylcholine.

GABAergic modulation of neurons in the nucleus of the solitary tract with ascending projections to the subfornical organ in the rat

Twenty-five neurons in the region of the nucleus of the solitary tract (NTS) were antidromically activated by electrical stimulation of the subfornical organ (SFO) in male rats under urethane anesthesia. Microiontophoretically applied bicuculline, a γ-aminobutyric acid (GABA) A antagonist, but not phaclofen, a GABA B antagonist, attenuated the post-antidromic inhibitory response evoked by SFO stimulation of approximately two-third ( n=17) of identified neurons, indicating the existence of recurrent inhibitory systems through GABA A receptors. Iontophoretically applied GABA decreased the spontaneous activity of all identified neurons, and the GABA-induced inhibition was prevented by simultaneously applied bicuculline, but not by phaclofen. Activation of peripheral baroreceptors, achieved by rising arterial blood pressure with an intravenous infusions of phenylepherine, suppressed the activity of the majority ( n=20) of identified neurons. The inhibitory response of identified neurons ( n=7) to baroreceptor activation was partially antagonized by iontophoretically applied bicuculline, but not by phaclofen. These results imply that GABAergic mechanisms may modulate the baroreceptor reflex acting on GABA A receptors of NTS neurons with ascending projections to the SFO in the region of the NTS.

Dipsogenic response induced by angiotensinergic pathways from the lateral hypothalamic area to the subfornical organ in rats

Experimental observations in several species have suggested that angiotensinergic neural circuits from the lateral hypothalamic area (LHA) to the subfornical organ (SFO) may participate in the control of drinking behavior in the rat. In an attempt to verify this possibility, experiments were undertaken to investigate whether activation of LHA neurons following microinjection of angiotensin II (ANG II) into the LHA elicits drinking. Injections of ANG II (10 −11 mol) into the LHA caused drinking in 25 out of 36 rats having the tips of cannulas in the LHA. The efficacy of ANG II was potentiated by increasing the dose of the drug. To clarify the contribution of angiotensinergic neurons in the LHA with efferent projections to the SFO to the drinking induced by ANG II, the effects of pretreatment with saralasin (Sar), a specific ANG II antagonist, in the SFO or its surrounding region on the drinking to ANG II were examined. Previous injections of Sar into the SFO significantly reduced the water intake caused by ANG II injected into the LHA, whereas treatment with Sar in the ventral hippocampal commissure (VHC) or third ventricle (3V) was without effect. These findings provide the evidence for the involvement of the angiotensinergic pathways from the LHA to the SFO in the dipsogenic action.

Angiotensinergic and noradrenergic mechanisms in the hypothalamic paraventricular nucleus participate in the drinking response induced by activation of the subfornical organ in rats

The present study was done to investigate the contribution of the hypothalamic paraventricular nucleus (PVN) to the drinking response caused by activation of the subfornical organ (SFO) following angiotensin II (ANG II) injections in the awake rat. Microinjection of ANG II into the SFO elicited the drinking response. Previous injections of either saralasin, an ANG II antagonist, or phentolamine, an α-adrenoceptor antagonist, bilaterally into the PVN resulted in the significant attenuation of the drinking response to ANG II. Similar injections of any of the β-adrenoceptor antagonist timolol, the muscarinic antagonist atropine, or saline vehicle into the PVN had no significant effect on the drinking response. In an attempt to clarify the neural mechanisms in the PVN involved in the drinking response to ANG II injected into the SFO, the effect of microinjection of ANG II into the SFO on noradrenaline (NA) release in the PVN was examined using intracerebral microdialysis techniques. The injection of the ANG II, but not saline vehicle, significantly enhanced the NA release in the region of the PVN. These results indicate the involvement of both the angiotensinergic and α-adrenergic systems in the PVN in the drinking response caused by angiotensinergic activation of the SFO, and imply that the angiotensinergic projections from the SFO to the PVN may serve to increase NA release which results in mediating water intake.

Activation of metabotropic glutamate receptors inhibits GABAergic transmission in the rat subfornical organ

Glutamate is known to increase neuronal excitability in the subfornical organ, a circumventricular organ devoid of the blood–brain barrier. To understand the synaptic mechanism of neuronal excitation by glutamate in this nucleus, we examined the effects of glutamate on GABAergic spontaneous inhibitory postsynaptic currents recorded from subfornical organ neurons in the rat brain slice. The baseline frequency, amplitude and decay time-constant of such spontaneous synaptic currents were 5.60 Hz, 119 pA and 17.3 ms, respectively. Glutamate (10–1000 μM) selectively inhibited the frequency of spontaneous GABAergic inhibitory postsynaptic currents (half-maximal effective concentration=47 μM) with little effects on their amplitudes and decay time constants. The inhibitory effect of glutamate on the frequency of spontaneous GABAergic postsynaptic currents was not blocked by tetrodotoxin (1 μM), or by the antagonists of ionotropic glutamate receptors. In contrast, such inhibitory effect of glutamate was mimicked by general or group II selective metabotropic glutamate receptor agonists such as DCGIV (2S,1′R,2′R,3′R)-2-(2′,3′-dicarboxycyclopropyl)glycine (half-maximal effective concentration=112 nM), but not by the agonists for group I or group III metabotropic glutamate receptors. Under current clamp mode, glutamate reduced the frequencies of spontaneous inhibitory postsynaptic potentials and action potentials in subfornical organ neurons. Our data indicate that glutamate decreases the frequency of spontaneous inhibitory postsynaptic currents by acting on the group II metabotropic glutamate receptors on axonal terminals in the subfornical organ. From these results we suggest that the glutamate-induced modulation of tonic GABAergic inhibitory synaptic activity can influence the excitability of subfornical organ neurons.

A subthreshold persistent sodium current mediates bursting in rat subfornical organ neurones.

It is widely accepted that while release of amino acid neurotransmitters occurs with relatively high fidelity, peptidergic synapses require clustered bursts of action potentials for optimal transmitter release. Here we describe for the first time the occurrence and mechanisms of bursting by neurones in the subfornical organ (SFO), cells that utilize the peptide angiotensin II (ANG) in neurotransmission in autonomic pathways. In current clamp recording of isolated SFO neurones in vitro, 53 % (n = 74) showed either spontaneous or evoked burst-like discharge patterns. Bursts typically appeared as shifts in bistable membrane potential, with action potentials superimposed on a depolarizing afterpotential (DAP). Similarly, in vivo single unit recordings of identified SFO neurones showed that 9 of 15 neurones fired in bursts. The pattern of bursting, as well as duration of evoked DAPs was strongly dependent upon membrane potential, suggesting that the DAP contributes to burst generation. Based on our previous observation of calcium-sensing receptor (CaR)-activated bursts, we investigated the effects of NPS R-467, an allosteric agonist of the CaR, on evoked DAPs. NPS R-467 (1 microM) potentiated DAP duration throughout the voltage range tested. We observed a dependence of evoked DAPs upon Na+ channels, as shown by sensitivity to tetrodotoxin (0.5 microM) or reduction of external [Na+] from 140 to 40 mM. The duration of DAPs suggested that a persistent Na+ current mediates these events. Voltage-clamp analysis revealed the presence of a subthreshold sodium current, INaP. Pharmacological blockade of INaP with 100 microM lidocaine reduced the duration of evoked DAPs, and inhibited bursting in SFO neurones. Facilitation of INaP with 10 nM anemone toxin (ATX) increased DAP duration and led to marked excitation of bursting cells. These data indicate that INaP is the main current underlying bursting in SFO neurones. Our observations of receptor-mediated facilitation of bursting by SFO neurones represents an intriguing mechanism through which the release of the peptide neurotransmitter ANG may be regulated.

Astrocytes in sensory circumventricular organs of the rat brain express functional binding sites for endothelin

Sensory circumventricular organs bordering the anterior third cerebral ventricle, the subfornical organ and the organum vasculosum laminae terminalis, lack blood–brain barrier characteristics and are therefore accessible to circulating peptides like endothelins. Astrocytes of the rat subfornical organ and organum vasculosum laminae terminalis additionally showed immunocytochemical localization of endothelin-1/endothelin-3-like peptides, possibly acting as circumventricular organ-intrinsic modulators. Employing [ 125I]endothelin-1 as radioligand, quantitative autoradiography demonstrated specific binding sites throughout the rat organum vasculosum laminae terminalis and subfornical organ, and competitive displacement studies revealed expression of both ET A and ET B receptor subtypes for either circumventricular organ. ET B receptor binding prevailed for the whole brain and ET A receptors could be labelled in the peripheral vascular system. To characterize endothelin-specific receptors in astrocytes of both circumventricular organs, alterations in the intracellular calcium concentration due to endothelin-1/endothelin-3 stimulation were studied in primary culture of subfornical organ and organum vasculosum laminae terminalis cells obtained from early postnatal rat pups. Endothelin-1 and endothelin-3 induced Ca 2+ transients in 9–13% of either subfornical organ or organum vasculosum laminae terminalis astrocytes, respectively, and some glial cells (subfornical organ: 2%, organum vasculosum laminae terminalis: 5%) responded to both endothelin analogues. The antagonistic action of BQ123 specific for ET A receptors (74% of all astrocytes tested), and the pronounced responsiveness to the ET B receptor agonist [4Ala]ET-1 (subfornical organ: 27%, organum vasculosum laminae terminalis: 35%) demonstrated glial expression of both endothelin receptor subtypes. Agonist-induced elevations in the intracellular calcium concentration proved to be independent of extracellular Ca 2+. In summary, the results indicate that endothelin(s) interact(s) with circumventricular organ astrocytes. Competitive receptor binding techniques using brain tissue sections as well as a fura-2 loaded primary cell culture system of the subfornical organ and organum vasculosum laminae terminalis demonstrate glial expression of functional ET A and ET B receptors, with calcium as intracellular messenger emerging primarily from intracellular stores. Endothelin(s) of both circulating and circumventricular organ-intrinsic origin may afferently transfer information important for cardiovascular homeostasis to circumventricular organs serving as “windows to the brain”.

Involvement of angiotensinergic and noradrenergic systems in the hypothalamic paraventricular nucleus in the drinking response induced by activation of the subfornical organ in rats

Involvement of angiotensinergic and noradrenergic systems in the hypothalamic paraventricular nucleus in the drinking response induced by activation of the subfornical organ in rats

Neuronal actions of oxytocin on the subfornical organ of male rats.

The aim of this study was to investigate effects of oxytocin (OT) on electrical neuronal activities in rat subfornical organ (SFO) and compare its action with the well-described excitatory effects of blood-borne angiotensin II (ANG II) on the same SFO neurons. With the use of extracellular recordings from spontaneously active neurons in slice preparations of the SFO of male rats, 11.7% of tested neurons (n = 206) were excited and 9.7% were inhibited by superfusion with 10(-6) M OT. Both excitatory and inhibitory effects of OT were dose dependent with similar threshold concentrations and were blocked by a specific OT-receptor antagonist but not by a vasopressin receptor antagonist. Blocking synaptic transmission with low calcium medium suppressed only inhibitory effects of OT. All but one of the OT-sensitive neurons were also excited by superfusion with ANG II at a concentration much lower than required for OT, suggesting that synaptically released OT rather than blood-borne OT alters the activity of SFO neurons in vivo. The results support the hypothesis that neurally released OT may modulate SFO-mediated functions by acting on OT-sensitive neurons.

Functional evidence for subfornical organ-intrinsic conversion of angiotensin I to angiotensin II.

Using extracellular electrophysiological recording in an in vitro slice preparation, we investigated whether ANG I can be locally converted to the functionally active ANG II within the rat subfornical organ (SFO). ANG I and ANG II (10(-8)-10(-7) M) excited approximately 75% of all neurons tested with both peptides (n = 25); the remainder were insensitive. The increase in firing rate and the duration and the latency of the responses of identical neurons, superfused with equimolar concentrations of ANG I and ANG II, were not different. The threshold concentrations of the ANG I- and ANG II-induced excitations were both 10(-9) M. Inhibition of the angiotensin-converting enzyme by captopril (10(-4) M; n = 8) completely blocked the ANG I-induced excitation, a 10-fold lower dose was only effective in two of four neurons. The AT1-receptor antagonist losartan (10(-5) M; n = 6) abolished the excitation caused by ANG I and ANG II. Subcutaneous injections of equimolar doses of ANG I and ANG II (200 microliters; 2 x 10(-4) M) in water-sated rats similarly increased water intake by 2.4 +/- 0.5 (n = 16) and 2. 7 +/- 0.4 ml (n = 20) after 1 h, respectively. Control rats receiving saline drank 0.07 +/- 0.06 ml under these conditions. Pretreatment with a low dose of captopril (2.3 x 10(-3) M) 10 min before the injection of ANG I caused a water intake of 2.8 +/- 0.5 ml (n = 10), whereas a high dose of captopril (4.6 x 10(-1) M) suppressed the dipsogenic response of ANG I entirely (n = 11). These data provide direct functional evidence for an SFO-intrinsic renin-angiotensin system (RAS) and underline the importance of the SFO as a central nervous interface connecting the peripheral with the central RAS.

Actions of amylin on subfornical organ neurons and on drinking behavior in rats

Amylin, a peptide hormone secreted by pancreatic beta-cells after food intake, contributes to metabolic control by regulating nutrient influx into the blood, whereas insulin promotes nutrient efflux and storage. We now report that amylin activates neurons in the subfornical organ (SFO), a structure in which the lack of a functional blood-brain barrier and the presence of a high density of amylin receptors may render it accessible and sensitive to circulating amylin. In an in vitro slice preparation of the rat SFO, 73% of 78 neurons were excited by superfusion with rat amylin (10(-8)-10(-7) M); the remainder were insensitive. The threshold concentration for the excitatory response of amylin was <10(-8) M and thus similar in potency to a previously reported excitatory effect of ANG II on the same neurons. The excitatory effect of amylin was completely blocked by coapplication of the selective amylin receptor antagonist AC-187 (10(-6)-10(-5) M) but was not affected by losartan (10(-5) M). Subcutaneous injections of 40 nmol of amylin significantly increased water intake in euhydrated rats, as did an equimolar dose of ANG II, which is a well-described SFO-mediated effect of circulating ANG II. These results point to the SFO as a sensory central nervous target for amylin released systemically in response to metabolic changes. Furthermore, we suggest that amylin release during food intake may stimulate prandial drinking.

Differential regulation of angiotensinogen and AT 1A receptor mRNA within the rat subfornical organ during dehydration

The present study describes the differential rostro-caudal patterning of angiotensinogen (AoGen) and AT 1A receptor mRNAs in the rat SFO using specific and validated oligodeoxynucleotide probes for in situ hybridization. Highest levels of AoGen-specific gene expression were observed in the rostral region of the SFO with gradually decreasing intensity towards the caudal region of this sensory circumventricular organ lacking blood–brain barrier function. AoGen-related hybridization signals proved to be specifically prominent above cells in lateral aspects of the SFO, surrounding septal venules. Maximal expression of the AT 1A receptor-specific gene, on the other hand, could be detected in the neuron-enriched, ventro-medial core region and dorsal annulus of the SFO, with low-intensity hybridization signals in its rostral and caudal parts. Water deprivation for 48 h, leading to extracellular hypertonic hypovolemia with elevated circulating AngII concentrations within the physiological range, caused a significant increase in AoGen-specific hybridization signals in the rostral and medial SFO regions. AT 1A receptor gene expression and AngII receptor binding were markedly stimulated in the medial and caudal regions of the SFO (core and annulus) as compared to euhydrated animals. These data indicate, that mild dehydration differentially up-regulates AoGen- and AT 1A receptor-specific mRNA formation as well as AT 1 receptor binding in distinct regions of the SFO, and supports the involvement of different cellular subgroups in the expression of two major components of the central nervous renin-angiotensin system in this sensory circumventricular organ.

Multiple receptor subtypes mediate the effects of serotonin on rat subfornical organ neurons.

The subfornical organ (SFO) receives significant serotonergic innervation. However, few reports have examined the functional effects of serotonin on SFO neurons. This study characterized the effects of serotonin on spontaneously firing SFO neurons in the rat brain slice. Of 31 neurons tested, 80% responded to serotonin (1-100 microM) with either an increase (n = 15) or decrease (n = 10) in spontaneous activity. Responses to serotonin were dose dependent and persisted after synaptic blockade. Excitatory responses could also be mimicked by the 5-hydroxytryptamine (5-HT)2A/2C receptor agonist 2,5-dimethoxy-4-iodoamphetamine (DOI; 1-10 microM) and could be blocked by the 5-HT2A/2C-receptor antagonist LY-53,857 (10 microM). LY-53,857 unmasked inhibitory responses to serotonin in 56% of serotonin-excited cells tested. Serotonin-inhibited cells were also inhibited by the 5-HT1A-receptor agonist 8-hydroxy-2(di-n-propylamino)tetralin (8-OH-DPAT; 1-10 microM; n = 7). The data indicate that SFO neurons are responsive to serotonin via postsynaptic activation of multiple receptor subtypes. The results suggest that excitatory responses to serotonin are mediated by 5-HT2A or 5-HT2C receptors and that inhibitory responses may be mediated by 5-HT1A receptors. In addition, similar percentages of serotonin-excited and -inhibited cells were also sensitive to ANG II. As such the functional relationship between serotonin and ANG II in the SFO remains unclear.

Effect of calcitonin on the activity of ANG II-responsive neurons in the rat subfornical organ.

In addition to the well-documented ability of calcitonin to lower blood calcium levels, blood-borne calcitonin may also affect neurons located outside the blood-brain barrier, e.g., in the subfornical organ (SFO), where numerous receptors for this peptide have been described. In an in vitro preparation of the rat SFO, calcitonin activated 61% of 36 neurons, only 1 neuron was inhibited, and the remainder were unresponsive. All but two of the neurons excited by 10(-7) M calcitonin were also stimulated by 10(-7) M ANG II. The threshold concentration for the excitatory effects of calcitonin was 10(-9) M and was thus similar to ANG II. Like ANG II, subcutaneous injection of calcitonin stimulated water intake, although to a lower extent. These results suggest that blood-borne calcitonin could stimulate drinking by its excitatory effect on neurons in the SFO. Calcitonin, which is released during food intake, might be involved in prandial drinking, which is presently considered an acquired behavior.

Effect of glutamate and angiotensin II on whole cell currents and release of nitric oxide in the rat subfornical organ.

Blood-borne angiotensin II (AngII) is known to mediate water-intake by its excitatory effect on neurons in the subfornical organ (SFO). Conversely, nitric oxide (NO) has exclusively inhibitory effects on rat SFO-neurons and on SFO-mediated water-intake. Extracellular and patch-clamp recordings from freshly dissociated rat SFO-neurons showed that glutamate activates AngII-sensitive SFO-neurons by opening ligand-gated cation channels. An immunocytochemical study showed that activation of glutamate receptors increased the concentration of the inhibitory second messenger cGMP in the SFO. A model is proposed suggesting that NO protects SFO-neurons from overexcitability by excitatory neurotransmitters.

Ascending noradrenergic pathways from the nucleus of the solitary tract to the subfornical organ in rats

These studies examined the role of spinal N-methyl-D-aspartic acid (NMDA) receptors in mediating Sympathoexcitation evoked by stimulation of neurons in the rostral ventrolateral medulla (RVLM). In urethane-anesthetized rats, blood pressure, heart rate, and splanchnic sympathetic nerve activity (SNA) were recorded. The NMDA receptor antagonist D-2-amino-7-phosphonoheptanoic acid (D-AP7) was administered to the spinal cord via intrathecal (IT) catheter. Blockade of spinal NMDA receptors reduced arterial blood pressure, heart rate, and SNA. Spinal administration of D-AP7 markedly attenuated the pressor and sympathoexcitatory responses evoked by L-glutamate stimulation of the RVLM. The small increases in heart rate evoked by stimulation of the RVLM were not affected by IT administration of D-AP7. These results indicate that NMDA receptors in the spinal cord mediate the pressor and sympathoexcitatory responses evoked by activation of a bulbospinal pathway originating from the RVLM. Moreover, these data suggest that excitatory amino acid neurotransmitters and NMDA receptors in the spinal cord play an important role in the maintenance and regulation of SNA and cardiovascular function.

Heterogeneous actions of vasopressin on ANG II-sensitive neurons in the subfornical organ of rats

The aim of this study was to investigate the effects of the antidiuretic hormone arginine vasopressin (AVP), which is released in vivo during dehydration and hypovolemia to prevent further water loss, on the activity of neurons in the subfornical organ (SFO). The SFO is a brain structure with an open blood-brain barrier and is critically involved in angiotensin II (ANG II)-dependent water intake. SFO neurons were recorded extracellularly in tissue slices of the rat brain and were tested for responsiveness to AVP and ANG II. About one-half of 159 neurons tested with an AVP concentration of 10(-6) M in the superfusion medium were responsive, and approximately equal proportions were excited and inhibited. Neurons exhibiting the different response types did not differ from each other with respect to spontaneous discharge rate, latency, and duration of the response. Excitatory and inhibitory responses to AVP were dose dependent and reversible, and their threshold concentrations (10(-8) to 10(-9) M) were similar. Superfusion with a medium low in Ca2+ and high in Mg2+ showed that the excitatory effect is most likely direct, whereas the inhibitory effect largely depends on inhibitory synaptic interaction. About one-half of the SFO neurons excited by ANG II (10(-7) M) were responsive to AVP (10(-6) M), and equal proportions were inhibited and excited. Both excitatory and inhibitory AVP actions were blocked by the V1-receptor antagonist, Manning compound, and neurons responsive to AVP did not respond to the V2-receptor agonist [deamino-Cys1,D-Arg8]vasopressin. It is concluded that AVP, probably released from synaptic terminals, may increase or decrease the activity of neurons in the SFO, many of which are activated by ANG II. In contrast to previous experiments on ducks, in which the exclusively excitatory effect of the avian antidiuretic hormone arginine vasotocin on ANG II-sensitive SFO neurons correlates well with the dipsogenic effect of both peptides, a greater functional heterogeneity exists among AVP-responsive neurons in the rat SFO.