Neurons In Rostral Ventrolateral Medulla Research Articles

Is the RVLM a key site for sex-related differences in blood pressure regulation?Focus on “Sex differences in angiotensin signaling in bulbospinal neurons in the rat rostral ventrolateral medulla,” by Wang et al.

it is well known that arterial blood pressure and the prevalence of hypertension and other cardiovascular diseases is higher in men than in age-matched premenopausal women, although after menopause these differences are reduced or even reversed ([10][1]). It is likely that many factors contribute to

Sex differences in angiotensin signaling in bulbospinal neurons in the rat rostral ventrolateral medulla

Sex differences may play a significant role in determining the risk of hypertension. Bulbospinal neurons in the rostral ventrolateral medulla (RVLM) are involved in the tonic regulation of arterial pressure and participate in the central mechanisms of hypertension. Angiotensin II (ANG II) acting on angiotensin type 1 (AT(1)) receptors in RVLM neurons is implicated in the development of hypertension by activating NADPH oxidase and producing reactive oxygen species (ROS). Therefore, we analyzed RVLM bulbospinal neurons to determine whether there are sex differences in: 1) immunolabeling for AT(1) receptors and the key NADPH oxidase subunit p47 using dual-label immunoelectron microscopy, and 2) the effects of ANG II on ROS production and Ca(2+) currents using, respectively, hydroethidine fluoromicrography and patch-clamping. In tyrosine hydroxylase-positive RVLM neurons, female rats displayed significantly more AT(1) receptor immunoreactivity and less p47 immunoreactivity than male rats (P < 0.05). Although ANG II (100 nM) induced comparable ROS production in dissociated RVLM bulbospinal neurons of female and male rats (P > 0.05), an effect mediated by AT(1) receptors and NADPH oxidase, it triggered significantly larger dihydropyridine-sensitive long-lasting (L-type) Ca(2+) currents in female RVLM neurons (P < 0.05). These observations suggest that an increase in AT(1) receptors in female RVLM neurons is counterbalanced by a reduction in p47 levels, such that ANG II-induced ROS production does not differ between females and males. Since the Ca(2+) current activator Bay K 8644 induced larger Ca(2+) currents in females than in male RVLM neurons, increased ANG II-induced L-type Ca(2+) currents in females may result from sex differences in calcium channel densities or dynamics.

Role of the caudal pressor area in the regulation of sympathetic vasomotor tone

It is well known that the ventrolateral medulla contains neurons involved in the tonic and reflex control of the cardiovascular system. Two regions within the ventrolateral medulla were initially identified: the rostral ventrolateral medulla (RVLM) and the caudal ventrolateral medulla (CVLM). Activation of the RVLM raises arterial blood pressure and sympathetic nerve activity, and activation of the CVLM causes opposite effects. The RVLM premotor neurons project directly to sympathetic preganglionic neurons and are involved in the maintenance of resting sympathetic vasomotor tone. A significant proportion of tonic activity in the RVLM sympathetic premotor neurons is driven by neurons located in a third region of the ventrolateral medulla denominated caudal pressor area (CPA). The CPA is a pressor region located at the extreme caudal part of the ventrolateral medulla that appears to have an important role controlling the activity of RVLM neurons. In this brief review, we will address the importance of the ventrolateral medulla neurons for the generation of resting sympathetic tone related to arterial blood pressure control focusing on two regions, the RVLM and the CPA.

Lesions of the Ventral Lamina Terminalis Prevent Salt‐Induced Changes in the Responsiveness of Neurons in the Rostral Ventrolateral Medulla (RVLM)

Sympathetic nerve activity (SNA) and arterial blood pressure (ABP) responses to injection of L‐glutamate or GABA into the RVLM are enhanced during elevated dietary salt intake. The present study determined whether sodium‐sensing neurons of the forebrain lamina terminalis mediate these changes. Male Sprague‐dawley rats with or without lesions of the ventral lamina terminalis were given access to normal chow and either water or 0.9% NaCl for 14 days. Injection of L‐glutamate into the RVLM of control rats drinking 0.9% NaCl (57±2 ml/day, n=13) versus water (35±1 ml/day, n=11) produced greater changes in renal SNA (0.1nmol: 81±5 vs 52±5%; 1nmol: 123±8 vs 82±6%) and ABP (0.1nmol: 23±2 vs 10±1; 1nmol: 44±3 vs 24±1 mmHg). In marked contrast, injection of L‐glutamate in lesioned rats drinking 0.9% NaCl (56±7 ml/day, n=5) versus water (42±2 ml/day, n=5) showed similar increases in renal SNA (0.1nmol: 62±9 vs 57±11%; 1nmol: 87±4 vs 85±17%) and ABP (0.1nmol: 15±4 vs 15±3; 1nmol: 29±4 vs 31±5 mmHg). Similarly, injection of GABA produced greater depressor responses in control rats drinking 0.9% NaCl versus water (0.1nmol: −15±1 vs −7±1; 10nmol: −31±3 vs −15±1 mmHg, n&gt;7), and this effect was prevented in lesioned rats (0.1nmol: −6±1 vs −7±1; 10nmol: −12±2 vs −15±1 mmHg, n=3). These findings suggest that increased dietary salt activates neurons in the forebrain lamina terminalis to enhance the responsiveness of RVLM neurons.

Catecholamine neurons in the rostral ventrolateral medulla (RVLM) contribute to the cardiovascular effects of endotoxin challenge

The rostral ventrolateral medulla (RVLM) is critical to neural control of cardiovascular function. Spinally‐projecting sympathoexcitatory RVLM neurons can be divided into two groups: C1 neurons that express catecholamine biosynthetic enzymes, and non‐C1 neurons that do not. Selective lesioning of RVLM‐C1 neurons in rats with saporin conjugated to an antibody against dopamine beta‐hydroxylase has provided evidence that C1 neurons contribute to cardiovascular reflexes, homeostatic responses to glucoprivation, pressor responses to restraint stress, and other sympathoexcitatory responses. We hypothesized that C1 neurons also play a role in cardiovascular responses to interoceptive stress induced by injection of endotoxin (lipopolysaccharide; LPS), a fragment of gram‐negative bacterial cell walls that presents an immune challenge by stimulating the release of pro‐inflammatory cytokines. Radiotelemetry transmitters were used to record blood pressure (BP) and heart rate (HR) in conscious rats after LPS challenge (200 ug/kg, i.p.). In rats with substantial loss of C1 neurons, BP was attenuated throughout the biphasic pressor response to LPS, and both HR and HR variability response patterns were altered. These data provide further support for an important role of RVLM‐C1 neurons in mediating sympathoexcitatory cardiovascular responses to homeostatic challenges. (Supported by HL55786 and MH59911).

Monosynaptic Excitatory Connection from the Rostral Ventrolateral Medulla to Sympathetic Preganglionic Neurons Revealed by Simultaneous Recordings

To directly investigate whether a monosynaptic connection exists between neurons in the rostral ventrolateral medulla (RVLM) and sympathetic preganglionic neurons (SPNs), we used simultaneous extracellular recordings of RVLM neurons and whole-cell patch-clamp recordings of SPNs at the Th2 level and analyzed them by spike-triggered averaging. We averaged 200 sweeps of membrane potentials in SPN triggered by the spikes in the RVLM neuron. No clear postsynaptic potentials were detected in the averaged wave of SPNs before angiotensin II (Ang II) superfusion, whereas during superfusion with Ang II (6 micromol/L) on the medulla oblongata side alone excitatory postsynaptic potentials (EPSPs) were clearly found in the SPN of 3 out of 10 pairs at 40 +/- 1 ms after the averaged triggering spike in the RVLM neuron. We consider them to be monosynaptic EPSPs, because 1) the averaged EPSPs exhibited a sharp rise time, 2) the onset latency of the averaged EPSPs in the SPNs after the trigger spike in the RVLM was the same as the latency of the antidromic action potentials in the RVLM neurons in response to electrical stimulation of the SPNs, and 3) the amplitude of the averaged EPSPs was over 2 mV. In summary, combining simultaneous recording and spike-triggered averaging allowed us to demonstrate a monosynaptic excitatory connection between a single RVLM neuron and a single SPN in the thoracic spinal cord. Such connections provide the basis for the maintenance of sympathetic tone and the integrative reflex that relays through the RVLM. The results explain the mechanism by which Ang II in the RVLM area increases peripheral sympathetic activity and blood pressure.

Inhibition of Nischarin Expression Attenuates Rilmenidine-Evoked Hypotension and Phosphorylated Extracellular Signal-Regulated Kinase 1/2 Production in the Rostral Ventrolateral Medulla of Rats

Imidazoline (I(1))-evoked hypotension is linked to enhanced phosphorylated extracellular signal-regulated kinase (pERK)1/2 production in the rostral ventrolateral medulla (RVLM). Recent cell culture findings suggest that nischarin is a candidate for the I(1) receptor. In the present study, nischarin antisense oligodeoxynucleotide (ODN) (AS1 or AS2), designed according to nischarin cDNA sequence, was administered intracisternally (i.c., 2 nmol/rat for 2 days) to knockdown central nischarin expression; control rats received the corresponding mismatched ODN (MM1 or MM2) or artificial cerebrospinal fluid (aCSF). We investigated the effects of AS1 or AS2 on nischarin expression in the RVLM, and on the hypotension and RVLM pERK1/2 production elicited by the I(1)-selective agonist rilmenidine (25 mug/rat i.c.). Compared with aCSF, the mismatched ODN (MM1 or MM2) had no significant effect on RVLM nischarin expression or the cardiovascular and cellular (RVLM pERK1/2) responses elicited by rilmenidine. However, either antisense ODN substantially (>80%) reduced nischarin expression in the RVLM (AS1/MM1, 3 +/- 1 versus 32 +/- 2 positive cells; AS2/MM2, 4 +/- 1 versus 31 +/- 2 positive cells) and abrogated rilmenidine (I(1))-evoked hypotension (AS1/MM1, -4.1 +/- 0.9 versus -10.8 +/- 1.9 mm Hg; AS2/MM2, -2.1 +/- 1.1 versus -15.3 +/- 2.5 mm Hg) and ERK1/2 activation in the RVLM (AS1/MM1, 10 +/- 1 versus 15 +/- 2 positive cells; AS2/MM2, 9 +/- 1 versus 18 +/- 2 positive cells). Finally, pERK1/2 generated by central I(1) receptor activation is colocalized with nischarin in the RVLM neurons. This is the first evidence in vivo that nischarin plays a critical role in I(1) receptor-mediated pERK1/2 production in the RVLM and the subsequent hypotension.

Anatomical evidence of regional specific effects of acupuncture on gastric motor function in rats

To obtain the anatomical evidences of possible neural pathways in mediating acupuncture-induced gastric motor responses, we studied c-Fos immunohistochemistry of the brain stem in response to acupuncture in rats. Acupuncture needles were inserted at the bilateral acupoints of ST-36 (lower limb) or ST-25 (abdomen) for 30 min. After acupuncture, the brainstem was removed for c-Fos immunohistochemistry. The total number of c-Fos immunopositive cells was counted in the dorsal motor nucleus of the vagus (DMV), the nucleus tractus solitarius (NTS) and the rostral ventrolateral medulla (RVLM). Acupuncture at ST-36, but not ST-25, significantly increased the number of c-Fos immunopositive cells at the DMV to 6.7 +/− 0.4 cells/section, compared to that of controls (1.7 +/− 0.2 cells/section) ( n = 5, P < 0.05). Acupuncture at ST-25, but not ST-36, significantly increased the number of c-Fos immunopositive cells at the RVLM to 12.6 +/− 0.8 cells/section, compared to that of controls (4.2 +/− 0.7 cells/section) ( n = 5, P < 0.05). Acupuncture at ST-36 also increased the number of c-Fos immunopositive cells at the medio-caudal and caudal NTS. On the other hand, acupuncture at ST-25 increased the number of c-Fos immunopositive cells at the medio-caudal NTS. It is suggested that somatic afferents activated by acupuncture at ST-36 is conveyed to the medio-caudal and caudal NTS and stimulates the DMV neurons. In contrast, somatic afferents activated by acupuncture at ST-25 is conveyed to the medio-caudal NTS and stimulates the RVLM neurons. The RVLM neurons are known as premotor sympatho-excitatory neurons that provide drive to the sympathetic preganglionic neurons in the intermediolateral nucleus of the spinal cord. Thus, acupuncture at ST-36 stimulates gastric motility via vagal efferents, while acupuncture at ST-25 inhibits gastric motility via sympathetic efferents in rats.

Increased Dietary Salt Enhances Sympathoexcitatory and Sympathoinhibitory Responses From the Rostral Ventrolateral Medulla

Increased dietary salt exaggerates arterial blood pressure (ABP) responses evoked from the rostral ventrolateral medulla (RVLM). The present study determined whether these enhanced pressor responses were directly attributable to a greater increase in sympathetic nerve activity (SNA) and whether these enhanced responses were balanced by a greater responsiveness of RVLM neurons to inhibitory input. Male Sprague-Dawley rats were fed normal chow and given access to either water or a 1% NaCl solution for 14 days. Injection of l-glutamate (0.03, 0.1, 1.0, and 3.0 nmol) into the RVLM produced a significantly greater increase in renal SNA, splanchnic SNA, and ABP in rats drinking 1% NaCl versus water. Conversely, injection of the inhibitory amino acid gamma-aminobutyric acid (0.1, 1.0, and 10 nmol) into the RVLM produced significantly greater decreases in renal SNA, splanchnic SNA, and ABP of rats drinking 1% NaCl versus water. These enhanced SNA and ABP responses to l-glutamate and gamma-aminobutyric acid were not observed in rats drinking 1% NaCl for 1 or 7 days but were present in rats drinking 1% NaCl for 21 days. Moreover, the dietary salt-induced enhancement of both sympathoexcitatory and sympathoinhibitory responses from the RVLM persisted after the 1% NaCl solution was replaced with water for 1, but not 7, days. These findings indicate that the potentiated ABP responses observed previously are mediated by parallel changes in SNA, and these responses depend on a slowly developing and reversible form of neuronal plasticity.

Transcriptional up-regulation of nitric oxide synthase II by nuclear factor-kappaB at rostral ventrolateral medulla in a rat mevinphos intoxication model of brain stem death.

As the origin of a 'life-and-death' signal that reflects central cardiovascular regulatory failure during brain stem death, the rostral ventrolateral medulla (RVLM) is a suitable neural substrate for mechanistic delineation of this vital phenomenon. Using a clinically relevant animal model that employed the organophosphate pesticide mevinphos (Mev) as the experimental insult, we evaluated the hypothesis that transcriptional up-regulation of nitric oxide synthase I or II (NOS I or II) gene expression by nuclear factor-kappaB (NF-kappaB) on activation of muscarinic receptors in the RVLM underlies brain stem death. In Sprague-Dawley rats maintained under propofol anaesthesia, co-microinjection of muscarinic M2R (methoctramine) or M4R (tropicamide), but not M1R (pirenzepine) or M3R (4-diphenylacetoxy-N-dimethylpiperidinium) antagonist significantly reduced the enhanced NOS I-protein kinase G signalling ('pro-life' phase) or augmented NOS II-peroxynitrite cascade ('pro-death' phase) in ventrolateral medulla, blunted the biphasic increase and decrease in baroreceptor reflex-mediated sympathetic vasomotor tone that reflect the transition from life to death, and diminished the elevated DNA binding activity or nucleus-bound translocation of NF-kappaB in RVLM neurons induced by microinjection of Mev into the bilateral RVLM. However, NF-kappaB inhibitors (diethyldithiocarbamate or pyrrolidine dithiocarbamate) or double-stranded kappaB decoy DNA preferentially antagonized the augmented NOS II-peroxynitrite cascade and the associated cardiovascular depression exhibited during the 'pro-death' phase. We conclude that transcriptional up-regulation of NOS II gene expression by activation of NF-kappaB on selective stimulation of muscarinic M2 or M4 subtype receptors in the RVLM underlies the elicited cardiovascular depression during the 'pro-death' phase in our Mev intoxication model of brain stem death.

Obesity‐induced hypertension depends on neurons of the rostral ventrolateral medulla (RVLM)

Obesity‐induced hypertension depends on neurons of the rostral ventrolateral medulla (RVLM)

Central chemoreceptors and sympathetic vasomotor outflow

The present study explores how elevations in brain P(CO(2)) increase the sympathetic nerve discharge (SND). SND, phrenic nerve discharge (PND) and putative sympathoexcitatory vasomotor neurons of the rostral ventrolateral medulla (RVLM) were recorded in anaesthetized sino-aortic denervated and vagotomized rats. Hypercapnia (end-expiratory CO(2) from 5% to 10%) increased SND (97 +/- 6%) and the activity of RVLM neurons (67 +/- 4%). Injection of kynurenic acid (Kyn, ionotropic glutamate receptor antagonist) into RVLM or the retrotrapezoid nucleus (RTN) eliminated or reduced PND, respectively, but did not change the effect of CO(2) on SND. Bilateral injection of Kyn or muscimol into the rostral ventral respiratory group (rVRG-pre-Botzinger region, also called CVLM) eliminated PND while increasing the stimulatory effect of CO(2) on SND. Muscimol injection into commissural part of the solitary tract nucleus (commNTS) had no effect on PND or SND activation by CO(2). As expected, injection of Kyn into RVLM or muscimol into commNTS virtually blocked the effect of carotid body stimulation on SND in rats with intact carotid sinus nerves. In conclusion, CO(2) increases SND by activating RVLM sympathoexcitatory neurons. The relevant central chemoreceptors are probably located within or close to RVLM and not in the NTS or in the rVRG-pre-Botzinger/CVLM region. RVLM sympathoexcitatory neurons may be intrinsically pH-sensitive and/or receive excitatory synaptic inputs from RTN chemoreceptors. Activation of the central respiratory network reduces the overall sympathetic response to CO(2), presumably by activating barosensitive CVLM neurons and inhibiting RTN chemoreceptors.

Role of ventrolateral medulla in generating the 10-Hz rhythm in sympathetic nerve discharge

We recorded changes in right inferior cardiac and either left inferior cardiac or left vertebral sympathetic nerve discharge (SND) produced by unilateral microinjections of GABA-A and excitatory amino acid (EAA) receptor antagonists into the ventrolateral medulla (VLM) of urethane-anesthetized, baroreceptor-denervated cats. Unilateral microinjections of GABA-A receptor antagonists, SR-95531 or bicuculline, into single tracks in VLM anywhere between 1 and 5 mm rostral to the obex eliminated or markedly reduced 10-Hz power in SND on both sides of the body. Low-frequency components (<6 Hz) of SND were unaffected. Complete blockade of the 10-Hz rhythm occurred with a dose of SR-95531 as low as 6.25 pmol in a 50-nl volume. Unilateral microinjections of the nonselective EAA receptor antagonist, kynurenate (KYN; 7.5 nmol), into the caudal or rostral VLM significantly reduced, but did not eliminate, 10-Hz SND ipsilateral to the injection sites, while 10-Hz SND contralateral to the injection sites was not significantly changed. These observations suggest that 1) GABAergic transmission in VLM is critical for generation of the 10-Hz rhythm, 2) the caudal and rostral portions of VLM act together to generate the 10-Hz rhythm, and 3) 10-Hz rhythm generation depends, at least in part, on tonic or phasic excitatory drive to GABAergic interneurons in caudal VLM and presympathetic neurons in rostral VLM. The data also suggest that pathways interconnecting the two halves of the brain stem play an important role in promoting 10-Hz rhythm generation.

Direct Evidence for Imidazoline (I1) Receptor Modulation of Ethanol Action on Norepinephrine‐Containing Neurons in the Rostral Ventrolateral Medulla in Conscious Spontaneously Hypertensive Rats

Enhancement of the rostral ventrolateral medulla (RVLM) presympathetic (norepinephrine, NE) neuronal activity represents a neurochemical mechanism for the pressor effect of ethanol. In this study, we tested the hypothesis that ethanol action on RVLM presympathetic neurons is selectively influenced by the signaling of the local imidazoline (I1) receptor. To support a neuroanatomical and an I1-signaling selectivity of ethanol, and to circumvent the confounding effects of anesthesia, the dose-related neurochemical and blood pressure effects of ethanol were investigated in the presence of selective pharmacological interventions that cause reduction in the activity of RVLM or nucleus tractus solitarius (NTS) NE neurons via local activation of the I1 or the alpha2-adrenergic receptor in conscious spontaneously hypertensive rats. Local activation of the I1 receptor by rilmenidine (40 nmol) or by the I1/alpha2 receptor mixed agonist clonidine (1 nmol), and local activation of the alpha2-adrenergic receptor (alpha2AR) by the pure alpha2AR agonist alpha-methylnorepinephrine (alpha-MNE, 10 nmol) caused reductions in RVLM NE, and blood pressure. Intra-RVLM ethanol (1, 5, or 10 microg), microinjected at the nadir of the neurochemical and hypotensive responses, elicited dose-dependent increments in RVLM NE and blood pressure in the presence of local I1--but not alpha2-receptor activation. Only intra-NTS alpha-MNE, but not rilmenidine or clonidine, elicited reductions in local NE and blood pressure; ethanol failed to elicit any neurochemical or blood pressure responses in the presence of local activation of the alpha2AR within the NTS. The findings support the neuroanatomical selectivity of ethanol, and support the hypothesis that the neurochemical (RVLM NE), and the subsequent cardiovascular, effects of ethanol are selectively modulated by I1 receptor signaling in the RVLM.

Central Command Regulation of Circulatory Function Mediated by Descending Pontine Cholinergic Inputs to Sympathoexcitatory Rostral Ventrolateral Medulla Neurons

Central command is a feedforward neural mechanism that evokes parallel modifications of motor and cardiovascular function during arousal and exercise. The neural circuitry involved has not been elucidated. We have identified a cholinergic neural circuit that, when activated, mimics effects on tonic and reflex control of circulation similar to those evoked at the onset of and during exercise. Central muscarinic cholinergic receptor (mAChR) activation increased splanchnic sympathetic nerve activity (SNA) as well as the range and gain of the sympathetic baroreflex via activation of mAChR in the rostral ventrolateral medulla (RVLM) in anesthetized artificially ventilated Sprague-Dawley rats. RVLM mAChR activation also attenuated and inhibited the peripheral chemoreflex and somatosympathetic reflex, respectively. Cholinergic terminals made close appositions with a subpopulation of sympathoexcitatory RVLM neurons containing either preproenkephalin mRNA or tyrosine hydroxylase immunoreactivity. M2 and M3 receptor mRNA was present postsynaptically in only non-tyrosine hydroxylase neurons. Cholinergic inputs to the RVLM arise only from the pedunculopontine tegmental nucleus. Chemical activation of this region produced increases in muscle activity, SNA, and blood pressure and enhanced the SNA baroreflex; the latter effect was attenuated by mAChR blockade. These findings indicate a novel role for cholinergic input from the pedunculopontine tegmental nucleus to the RVLM in central cardiovascular command. This pathway is likely to be important during exercise where a centrally evoked facilitation of baroreflex control of the circulation is required to maintain blood flow to active muscle.

Neurons of the rostral ventrolateral medulla (RVLM) contribute to sympathoexcitatory and pressor responses during acute stimulation of forebrain osmoreceptors

Acute stimulation of forebrain osmoreceptors increases sympathetic nerve activity (SNA) and arterial blood pressure (ABP). The present study sought to determine whether the sympathetic nervous system contributes to this pressor response and whether sympathetic-regulatory neurons of the RVLM mediate the increase in SNA and ABP. In urethane/chloralose anesthetized rats (n=5), intracarotid injection of hypertonic NaCl (150, 375, and 750 mM in 150 μl) dose-dependently increased mean ABP, and this was eliminated by administration of the ganglionic blocker chlorisondamine (2 mg/kg, iv) at 150mM (1±1 vs 1±0 mmHg), 375mM (16±3 vs 1±0 mmHg, p<0.05), and 750mM (27±4 vs 2±1 mmHg, p<0.01). In a second group (n=5), inhibition of the RVLM with microinjection of the GABAA receptor agonist muscimol (250 pmol/100 nl) significantly reduced the increase in mean ABP (150 mM: −3±2 vs 1±1 mmHg; 375mM: 14±5 vs 0±4 mmHg; 750mM: 31±5 vs 2±8 mmHg, p<0.05), renal SNA (150 mM: −1±2 vs −2±2%; 375mM: 50±18 vs −17±6%; 750mM: 80±13 vs −13±16%, p<0.05), and splanchnic SNA (150mM: −6±4 vs −1±1%; 375mM: 54±14 vs −5±14%; 750mM: 99±12 vs 14±20%, p<0.05). The present findings demonstrate that stimulation of forebrain osmoreceptors produces a sympathetically-mediated increase in ABP that depends on the tonic activity of RVLM neurons.

Transneuronal tracing of neural pathways that regulate hindlimb muscle blood flow

Despite considerable interest in the neural mechanisms that regulate muscle blood flow, the descending pathways that control sympathetic outflow to skeletal muscles are not adequately understood. The present study mapped these pathways through the transneuronal transport of two recombinant strains of pseudorabies virus (PRV) injected into the gastrocnemius muscles in the left and right hindlimbs of rats: PRV-152 and PRV-BaBlu. To prevent PRV from being transmitted to the brain stem via motor circuitry, a spinal transection was performed just below the L2 level. Infected neurons were observed bilaterally in all of the areas of the brain that have previously been shown to contribute to regulating sympathetic outflow: the medullary raphe nuclei, rostral ventrolateral medulla (RVLM), rostral ventromedial medulla, A5 adrenergic cell group region, locus coeruleus, nucleus subcoeruleus, and the paraventricular nucleus of the hypothalamus. The RVLM, the brain stem region typically considered to play the largest role in regulating muscle blood flow, contained neurons infected following the shortest postinoculation survival times. Approximately half of the infected RVLM neurons were immunopositive for tyrosine hydroxylase, indicating that they were catecholaminergic. Many (47%) of the RVLM neurons were dually infected by the recombinants of PRV injected into the left and right hindlimb, suggesting that the central nervous system has a limited capacity to independently regulate blood flow to left and right hindlimb muscles.

DIFFERENTIAL CONTROL OF CARDIAC AND SYMPATHETIC VASOMOTOR ACTIVITY FROM THE DORSOMEDIAL HYPOTHALAMUS

1. The dorsomedial hypothalamus (DMH) plays a crucial role in mediating the cardiovascular responses to different stressors, including acute psychological stress and cold stress. Activation of neurons in the DMH evokes increases in arterial pressure and in the activity of sympathetic nerves innervating the heart, blood vessels and brown adipose tissue. The descending pathways from the DMH to the spinal sympathetic outflow include synapses with neurons in medullary nuclei and possibly other brain stem regions. 2. Recent studies from our and other laboratories have indicated that neurons in the rostral ventrolateral medulla (RVLM) and in the region of the raphe pallidus (RP) in the medulla are important components of the descending pathways that mediate the cardiovascular response to activation of the DMH. Neurons in the RP primarily mediate the sympathetic cardiac components of the DMH-evoked response, whereas the RVLM neurons primarily mediate the sympathetic vasomotor component. 3. Activation of DMH neurons not only increases heart rate and sympathetic vasomotor activity, but also resets the baroreceptor reflex such that it remains effective, without any decrease in sensitivity, over a higher operating range of arterial pressure. 4. Activation of 5-hydroxytryptamine 5-HT(1A) receptors in the medulla oblongata leads to a selective suppression of cardiac and sympathetic vasomotor components of the DMH-evoked response, but does not affect sympathetic reflex responses evoked from baroreceptors or chemoreceptors. Thus, central 5-HT(1A) receptors modulate cardiovascular responses evoked from the DMH in a highly potent but selective fashion.

Central chemoreceptors and sympathetic vasomotor outflow

The present study explores how elevations in brain P(CO(2)) increase the sympathetic nerve discharge (SND). SND, phrenic nerve discharge (PND) and putative sympathoexcitatory vasomotor neurons of the rostral ventrolateral medulla (RVLM) were recorded in anaesthetized sino-aortic denervated and vagotomized rats. Hypercapnia (end-expiratory CO(2) from 5% to 10%) increased SND (97 +/- 6%) and the activity of RVLM neurons (67 +/- 4%). Injection of kynurenic acid (Kyn, ionotropic glutamate receptor antagonist) into RVLM or the retrotrapezoid nucleus (RTN) eliminated or reduced PND, respectively, but did not change the effect of CO(2) on SND. Bilateral injection of Kyn or muscimol into the rostral ventral respiratory group (rVRG-pre-Bötzinger region, also called CVLM) eliminated PND while increasing the stimulatory effect of CO(2) on SND. Muscimol injection into commissural part of the solitary tract nucleus (commNTS) had no effect on PND or SND activation by CO(2). As expected, injection of Kyn into RVLM or muscimol into commNTS virtually blocked the effect of carotid body stimulation on SND in rats with intact carotid sinus nerves. In conclusion, CO(2) increases SND by activating RVLM sympathoexcitatory neurons. The relevant central chemoreceptors are probably located within or close to RVLM and not in the NTS or in the rVRG-pre-Bötzinger/CVLM region. RVLM sympathoexcitatory neurons may be intrinsically pH-sensitive and/or receive excitatory synaptic inputs from RTN chemoreceptors. Activation of the central respiratory network reduces the overall sympathetic response to CO(2), presumably by activating barosensitive CVLM neurons and inhibiting RTN chemoreceptors.

Efferent projections of rat rostroventrolateral medulla C1 catecholamine neurons: Implications for the central control of cardiovascular regulation

A replication-defective lentivirus vector that expresses enhanced green fluorescent protein (EGFP) under the control of a synthetic dopamine-beta-hydroxylase (DbetaH) promoter was used to define efferent projections of C1 catecholamine neurons in rat rostral ventrolateral medulla (RVLM). EGFP expression was restricted to C1 neurons and filled their somatodendritic compartments and efferent axons 7-28 days after vector injection. This included the descending projections to thoracic spinal cord and a network in brainstem, midbrain, and diencephalon. In caudal brainstem, restricted terminal fields were present in the dorsal motor vagal complex, A1, raphe pallidus and obscurus, and marginal layer of ventrolateral medulla. Innervation of raphe nuclei was most dense at the level of RVLM, but rostral levels of pallidus were devoid of innervation. A sparse commissural projection to contralateral RVLM was observed, and pericellular arbors were present in the dorsal reticular formation among the projection pathway of catecholamine axons. Rostral brainstem contained a dense innervation of locus coeruleus and the nucleus subcoeruleus. A restricted innervation of the ventrolateral column of the periaqueductal gray distinguished the midbrain. Forebrain labeling was restricted to the diencephalon, where distinctive terminal fields were observed in the paraventricular thalamic nucleus; the lateral hypothalamic area; and the paraventricular, dorsomedial, supraoptic, and median preoptic nuclei of hypothalamus. Projection fibers also coursed through the tuberal hypothalamus into the median eminence. Collectively, these data demonstrate that RVLM C1 neurons modulate the activity of other central cell groups known to participate in the regulation of cardiovascular and autonomic function.