Search PubMed⌕ Search

Biomedical subjects

K M Spyer

Publications and source records attributed to K M Spyer.

At least 19 recordsLinked to original sources
PubMed2000

Sensing arterial CO(2) levels: a role for medullary P2X receptors.

ATP has been shown to act as an excitatory neurotransmitter in the central nervous system. In this review, evidence is presented to indicate that when ATP is micro-injected into the ventrolateral medulla (VLM) of the rat, changes in respiratory activity are elicited. These effects, and accompanying changes in heart rate and blood pressure are mediated by P2X purinoreceptors. Immunocytochemistry indicates a prevalence of P2X(2) and P2X(6) purinoreceptors in this region of the medulla. The P2 purinoceptor antagonists, suramin and PPADS blunt the respiratory responses to changes in arterial CO(2) levels when micro-injected into the VLM. This effect is shown electrophysiologically to be mediated by purinoreceptors located primarily on respiratory neurones of the VLM including the Bötzinger complex. As the effects of agonist activation of P2X(2) purinoceptors expressed in HEK293 cells and Xenopus oocytes are potentiated by lowering pH, these data imply that the central respiratory response to CO(2) depends in part on the pH sensitivity of purinoreceptors located on inspiratory neurones. The implications for respiratory activity and control are discussed.

Adenosine Triphosphate↗
PubMed2000

Coexpression of rat P2X2 and P2X6 subunits in Xenopus oocytes.

Transcripts for P2X(2) and P2X(6) subunits are present in rat CNS and frequently colocalize in the same brainstem nuclei. When rat P2X(2) (rP2X(2)) and rat P2X(6) (rP2X(6)) receptors were expressed individually in Xenopus oocytes and studied under voltage-clamp conditions, only homomeric rP2X(2) receptors were fully functional and gave rise to large inward currents (2-3 microA) to extracellular ATP. Coexpression of rP2X(2) and rP2X(6) subunits in Xenopus oocytes resulted in a heteromeric rP2X(2/6) receptor, which showed a significantly different phenotype from the wild-type rP2X(2) receptor. Differences included reduction in agonist potencies and, in some cases (e.g., Ap(4)A), significant loss of agonist activity. ATP-evoked inward currents were biphasic at the heteromeric rP2X(2/6) receptor, particularly when Zn(2+) ions were present or extracellular pH was lowered. The pH range was narrower for H(+) enhancement of ATP responses at the heteromeric rP2X(2/6) receptor. Also, H(+) ions inhibited ATP responses at low pH levels (<pH 6.3). The pH-dependent blocking activity of suramin was changed at this heteromeric receptor, although the potentiating effect of Zn(2+) on ATP responses was unchanged. Thus, the rP2X(2/6) receptor is a functionally modified P2X(2)-like receptor with a distinct pattern of pH modulation of ATP activation and suramin blockade. Although homomeric P2X(6) receptors function poorly, the P2X(6) subunit can contribute to functional heteromeric P2X channels and may influence the phenotype of native P2X receptors in those cells in which it is expressed.

Adenosine Triphosphate↗
PubMed2000

Inhibition of rat nucleus tractus solitarius neurones by activation of 5-HT2C receptors.

In vivo, nucleus tractus solitarius (NTS) neurones receiving monosynaptic vagal input and inactive intermediate neurones were inhibited by both DOI and a selective 5-HT2C receptor agonist, MK-212. Cells receiving a more polysynaptic input were excited by DOI and although MK-212 also excited a few of these cells, the majority of cells in these groups were unaffected by MK-212. The inhibitory, but not the excitatory actions of both MK-212 and DOI were prevented by a selective 5-HT2C receptor antagonist, RS-102221. In contrast, most dorsal vagal preganglionic neurones were unaffected by application of either DOI or MK-212, the few remaining cells being excited by both agonists. These data demonstrate that DOI-evoked inhibition of NTS cells activated by vagal afferent input and DOI-evoked excitation of vagal preganglionic neurones is mediated by 5-HT2C receptors.

Amphetamines↗
PubMed2000

P2X purinoceptor-mediated excitation of trigeminal lingual nerve terminals in an in vitro intra-arterially perfused rat tongue preparation.

A novel in vitro intra-arterially perfused adult rat tongue-nerve preparation was used to explore the possible actions of P2X purinoceptor agonists (ATP and alpha,beta-methylene ATP (alpha, beta-meATP)) on sensory nerve terminals innervating the rat tongue. We made whole-nerve recordings of the trigeminal branch of the lingual nerve (LN), which conducts general sensory information (pain, temperature, touch, etc.), and the chorda tympani (CT), which conducts taste information. Changes in LN and CT activity following intra-arterial application of P2X agonists were compared. In seven preparations, bolus close-arterial injection of ATP (30-3000 microM, 0.1 ml) or alpha,beta-meATP (10-300 microM, 0.1 ml) induced a rapid (< 1 s after injection), dose-related increase in LN activity that decayed within a few seconds. The minimal concentration of ATP (100 microM) required to elicit a response was about 10-fold higher than that of alpha,beta-meATP (10 microM). Bolus injection of ATP or alpha,beta-meATP induced a moderate decrease in firing frequency in three of seven CT preparations. LN responses to P2X agonists showed signs of rapid desensitisation with the peak frequency of discharge being smaller when the agonists were applied at short intervals. Suramin (200 microM) or PPADS (200 microM) applied by intra-arterial perfusion each antagonised the rapid increase in LN activity following application of alpha,beta-meATP (100 microM). Capsaicin (10 microM, 0.1 ml, n = 5 preparations) was injected intra-arterially to desensitise nociceptive fibres. This was found to block (n = 2) or greatly reduce (n = 3) the excitatory effects of alpha,beta-meATP (100 microM, 0.1 ml) on LN activity, implying that only capsaicin-sensitive nociceptive fibres in LN were responsive to P2X agonists. In contrast to the consistent excitatory responses in LN activity following fast application of P2X agonists as bolus, a variable and moderate change in discharge rate of LN and no change in CT activity (n = 5) was observed after applying ATP (100-300 microM, n = 21) or alpha,beta-meATP (100-300 microM, n = 14) by intra-arterial perfusion. The variable responses in LN activity to slow perfusion in contrast to close-arterial bolus injection are consistent with activation of the rapidly desensitising P2X3 receptors. In summary, ATP and alpha,beta-meATP preferentially activate general sensory afferent fibres (LN) but not taste fibres (CT). We suggest that the increase in whole-nerve activity of LN following application of P2X agonists represents activation of nociceptive fibres which possess P2X3 receptors. Our data indicate that ATP and P2X3 receptors may play a role in nociception, rather than taste sensation in the tongue.

Adenosine Triphosphate↗
PubMed2000

ATP as a mediator of mammalian central CO2 chemoreception.

1. A role for P2 purinoceptors in the chemosensory response of respiratory neurones localised in the ventrolateral medulla to changes in arterial CO2 levels was investigated in the anaesthetised rat. Extracellular recordings were made from different classes of respiratory neurone and the effects of P2 receptor blockade on CO2-evoked changes in activity investigated. 2. Increasing inspired CO2 excited 85 % of inspiratory neurones in the pre-Botzinger complex. In all cases, CO2-evoked excitation was blocked by ionophoretic application of the P2 receptor antagonists suramin (0.02 M) and pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS; 100 microM), but not the adenosine receptor antagonist 8-phenyltheophylline (8-PT; 100 microM). Suramin and PPADS often reduced ongoing activity, and blocked the excitatory effects of ATP. Inspiratory neurones were also excited by the P2X receptor agonist alphabeta-methyleneATP, suggesting a specific role for P2X receptors. 3. Sixty-six per cent of pre-inspiratory neurones were also excited by CO2. This effect was reduced or abolished by prior application of P2 receptor antagonists. Although post-inspiratory and expiratory neurones were excited by increasing levels of CO2, and also by ionophoretically applied ATP, the CO2-evoked effects were unaffected by P2 receptor blockade. 4. We suggest that ATP, possibly acting via P2X purinoceptors localised within the ventral respiratory group, is involved in central chemoreception. Specifically, these distinctive CO2-P2X-mediated actions were observed only in inspiratory neurones (incrementing inspiratory neurones and pre-inspiratory neurones), which appear to have purinoceptors with pH sensitivity that can account for the actions of CO2 in modifying ventilatory activity.

Adenosine Triphosphate↗
PubMed2000

Localization and action of adenosine A2a receptors in regions of the brainstem important in cardiovascular control.

In vitro autoradiography and central microinjections of a P1 adenosine A2a receptor antagonist have been employed to investigate a possible role for centrally located adenosine A2a receptors in modulation of the baroreceptor reflex. In vitro autoradiography using [125I]4-(2-[7-amino-2-[2-furyl][3,2,4]triazolol[2,3-a][1,3,5]tr iazin-5-yl-amino]ethyl)phenol ([125I]ZM241385), the high-affinity adenosine A2a receptor antagonist, revealed a heterogeneous distribution of adenosine A2a binding sites within the lower brainstem of the rat. Image analysis showed high levels of binding in rostral regions of both the nucleus tractus solitarius and the ventrolateral medulla. Intermediate levels of binding were observed in the commissural nucleus tractus solitarius and the dorsal vagal motor nucleus, with low levels of binding in caudal regions of the nucleus tractus solitarius and the ventrolateral medulla, and the hypoglossal nucleus. Unilateral microinjections of unlabelled ZM241385 into the nucleus tractus solitarius had no effect on baseline levels of arterial pressure, heart rate and phrenic nerve activity recorded in anaesthetized, artificially ventilated rats. However, microinjections of ZM241385 reduced the bradycardia evoked by stimulation of the ipsilateral aortic nerve. In contrast, ZM241385 had no effect on the depressor response or the reduction in phrenic nerve activity evoked by aortic nerve stimulation. Our results indicate that adenosine A2a binding sites are located in a number of brainstem regions involved in autonomic function, consistent with the idea that adenosine acts as a neuromodulator of a variety of cardiorespiratory reflexes. Specifically, the data support the hypothesis that adenosine A2a receptors located within the nucleus tractus solitarius are activated during baroreceptor stimulation and have an important modulatory role in the pattern of cardiovascular changes associated with this reflex.

Adenosine↗
PubMed1999

Central CO2 chemoreception: a mechanism involving P2 purinoceptors localized in the ventrolateral medulla of the anaesthetized rat.

1. The involvement of P2 purinoceptors in chemosensory function in the ventrolateral regions of the medulla oblongata was investigated in the anaesthetized rat. We have investigated the effect of antagonizing, or desensitizing, P2 receptors in the retrofacial area of the ventrolateral medulla on factors modifying respiratory activity. 2. Bilateral microinjection of suramin (50 nl, 0.02 M), a P2 purinoceptor antagonist, into the retrofacial area in the artificially ventilated rat reduced resting phrenic nerve discharge. It also markedly affected the response of the phrenic nerve to increases in arterial CO2. Under conditions of hyperoxic, hypocapnic apnoea, the mean threshold for inducing phrenic nerve activity was raised significantly (from an end-tidal CO2 of 2.5 % to 4.5 %, n = 9). 3. In addition, the slope of the respiratory response curve to increases in CO2 was reduced after suramin. A similar effect was observed after desensitization of certain P2X receptors with alphabeta-methyleneATP. As arterial levels of O2 were greater than 100 mmHg, and an equivalent pattern of response was observed in sino-aortically denervated and vagotomized animals, we believe any contribution of the peripheral chemoreceptors to be minimal. 4. Our data suggest that respiratory neurones within the retrofacial area (Botzinger complex) represent part of the central site of action of CO2 on respiration. Moreover, our observations lead us to suggest that CO2-evoked changes in respiration are mediated at least in part by P2X purinoceptors.

Adenosine Triphosphate↗
PubMed1999

A novel influence of adenosine on ongoing activity in rat rostral ventrolateral medulla.

We have investigated whether exogenously applied adenosine modulates neuronal activity in a region of the central nervous system crucial for cardiovascular regulation. Extracellular recordings were made from neurons in the rostral ventrolateral medulla of the anaesthetized rat. Ionophoretic application of adenosine altered ongoing activity in 91% of neurons, evoking either a long-lasting depression or a short-lasting increase in firing rate. Both responses were blocked by application of the broad spectrum adenosine receptor antagonist 8-sulphophenyltheophylline, indicating that the responses were mediated by specific cell surface receptors. The adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine blocked the increase, and partially blocked the decrease in firing rate in response to adenosine. The GABA(A) receptor antagonist bicuculline also blocked the increase in firing rate in response to adenosine, suggesting that adenosine may inhibit release of GABA from axon terminals in this region. The adenosine A2a receptor agonist CGS 21680 produced a long-lasting depression of ongoing activity. These results suggest that A1 receptors mediate an increase in firing rate, whilst A1 and A2a receptors mediate decreases in firing rate in some rostral ventrolateral medulla neurons. Thus, adenosine has been shown to modulate the ongoing activity of neurons in the rostral ventrolateral medulla by acting at both A1 and A2a receptors. Accordingly, we suggest, and provide some evidence to support the idea, that adenosine acts as an important neuromodulator in this region of the central nervous system, possibly by modulating the presynaptic release of neurotransmitters such as GABA.

Adenosine↗
PubMed1999

Late vagal inhibition in neurons of the ventrolateral medulla oblongata in the rat.

Stimulation of cervical vagal afferents elicits long-lasting inhibitory effects in a variety of neuronal populations, although little is known concerning the cellular mechanisms that are involved in these effects. In the present study, the electrophysiological characteristics of responses elicited by cumulative activation of vagal afferents were examined in neurons of the rostral ventrolateral medulla oblongata, which play an important role in the coordination of cardiovascular and other visceral activities. The study has focused on the late-onset, slow inhibitory component of vagal responses, which is likely to affect the temporal modulation of postsynaptic effects. Vagal stimulation elicited four distinct response patterns in intracellularly penetrated neurons (n = 78): excitation, inhibition, excitation-inhibition and inhibition-inhibition. The late inhibitory component was encountered in 43 (55%) of the cells, including five putative medullospinal neurons. It was due to a postsynaptic hyperpolarization which reversed at potentials more negative than -83 mV. The voltage dependency, as well as the average onset latency (93+/-3.0 ms), duration (270+/-16.5 ms) and amplitude (1.3+/-0.2 mV as measured at resting membrane potentials), of late inhibition were clearly different from those of the short-latency inhibitory response. The differences in the voltage dependency and time-course of the short-latency responses and the late inhibition indicate that they are mediated by different central relays. In the majority of neurons, late inhibition could be elicited by stimulating only myelinated vagal afferents. The magnitude of the response was, however, significantly enhanced in 63% of the examined cells when the intensity of stimulation was raised to recruit further myelinated and non-myelinated fibres. This indicates that late vagal inhibition is often elicited by a cumulative activation of convergent afferent inputs. The intracellularly labelled vagally responsive neurons were present at all rostrocaudal levels of the rostral ventrolateral medulla, with an accumulation in the region of the lateral paragigantocellular nucleus. Neurons that exhibited late vagal inhibition were dominant in the juxtafacial region of this nucleus. Due to its slow time-course, late vagal inhibition may contribute to a tonic modulation of the activity of neurons in the rostral ventrolateral medulla oblongata. It is proposed that late vagal inhibition plays an important role in the temporal integration of sensory inputs in neurons of the rostral ventrolateral medulla oblongata. The time-course and strength of this modulatory effect are related to the level of activity in those visceral sensory inputs that converge onto the inhibitory interneurons that mediate late inhibition to rostral ventrolateral medulla oblongata neurons.

Animals↗
PubMed1999

Vagal modulation of responses elicited by stimulation of the aortic depressor nerve in neurons of the rostral ventrolateral medulla oblongata in the rat.

Stimulation of cervical vagal afferents inhibits central sympathetic outflows in part by inhibiting the ongoing activity of putative baroreceptive neurons in the rostral ventrolateral medulla oblongata. The aim of the present study was to examine the electrophysiological characteristics of vagal responses and their interactions with responses elicited by stimulation of the aortic nerve in neurons there. The study focused on the role of the long-lasting, late-onset vagal inhibition, which is likely to play an important role in the tonic inhibitory effects of vagal afferent stimulation. In vivo intracellular recordings were obtained from 33 neurons that received convergent inputs from aortic and vagal afferents. Sixty-four percent of these neurons exhibited a late inhibition following electrical stimulation of myelinated vagal afferents (mean onset latency of 100+/-5 ms). The average duration of late inhibition (294+/-19 ms) exceeded the duration of the cardiac cycle. As a consequence of this, sustained vagal stimulation diminished the effect of rhythmic baroreceptor inputs in neurons that exhibited late vagal inhibition. Simultaneous activation of aortic and vagal afferents significantly increased the magnitude of late inhibition, even in those neurons where stimulation of the aortic nerve alone did not elicit a response (n = 15). This suggested that the convergence between vagal and aortic afferent inputs occurred in inhibitory inteneurons antecedent to the recorded rostral ventrolateral medulla oblongata neurons. Focal stimulation of the caudal part of the nucleus of the solitary tract also elicited a late-onset inhibition in 73% of the neurons that responded to stimulation of the aortic nerve. This inhibition appeared to be similar to the late vagal inhibition, except for its shorter average onset latency (64+/-7 ms). Based on this observation, it is proposed that inhibitory inteneurons that mediate late inhibition to rostral ventrolateral medulla oblongata neurons may lie within the caudal part of the nucleus of the solitary tract. The present study established that activation of myelinated vagal afferents exerts a complex modulation over the ongoing and evoked activity of neurons that respond to stimulation of the aortic nerve. The complex interaction that occurs between aortic and vagal inputs in neurons of the rostral ventrolateral medulla may be implicated in long-term modulation of sympathetic outflows in response to changes in the activation of visceral receptors supplied by vagus afferents. The modulation elicited by late vagal inhibition may help to adjust cardiovascular outflows according to requirements set by the thoraco-abdominal visceral environment.

Animals↗
PubMed1999

Characterization of P2 receptors modulating neural activity in rat rostral ventrolateral medulla.

This study investigated the effects of ATP, and related compounds, on the activity of neurons within the rostral ventrolateral medulla, an area of fundamental importance in reflex control of the cardiovascular system. Extracellular recordings were made from single neurons in anaesthetized, paralysed and artificially ventilated rats. Ionophoretic application of alpha,beta-methylene-ATP, adenosine 5'-O-(2-thiodiphosphate), UTP, 2-methylthio-ATP and ATP altered the ongoing activity in the majority of neurons (>74% of neurons), generally causing increases in the firing rate. Nine of 11 cells with presumed spinal projection were excited by ATP and/or the P2X-selective agonist alpha,beta-methylene-ATP. Desensitization of the excitatory responses to alpha,beta-methylene-ATP was observed in four of 20 rostral ventrolateral medulla neurons. For the remainder of the rostral ventrolateral medulla neurons, the increase in firing rate evoked by alpha,beta-methylene-ATP, and by the other purine compounds tested, did not undergo desensitization. Suramin, a P2 receptor antagonist, blocked excitatory responses to adenosine 5'-O-(2-thiodiphosphate) or alpha,beta-methylene-ATP in five of 16 neurons. These results indicate that ATP can modulate the activity of neurons in the rostral ventrolateral medulla via actions at P2 purine receptors. The data suggest that both P2X and P2Y receptors are involved, and that the functional expression of these receptors within the rostral ventrolateral medulla is not uniform.

Adenosine Diphosphate↗
PubMed1998

Convergence properties of solitary tract neurons responsive to cardiac receptor stimulation in the anesthetized cat.

The convergence pattern of cardiac receptors, pulmonary C-fibers, carotid chemoreceptor, and baroreceptor afferents onto neurons within the nucleus of the solitary tract (NTS) was studied in the anesthetized (pentobarbitone sodium, 40 mg/kg,) paralyzed and artificially ventilated cat. Extra- and intracellular recordings were made from NTS neurons while stimulating both cardiac receptors by aortic root injections of veratridine (1-3 micrograms/kg) and pulmonary C-fibers by a right atrial injection of phenylbiguanide (10-20 micrograms/kg). The ipsilateral carotid body was stimulated by using arterial injection of CO2-saturated bicarbonate solution, whereas inflation of the ipsilateral carotid sinus was used to activate baroreceptors. The ipsilateral cardiac vagal branch, cervical vagus, and carotid sinus nerves were stimulated electrically (1 Hz, 0.2-1 ms, 1-35 V). In 78 NTS neurons recorded either extracellularly (n = 47) or intracellularly (n = 31), electrical stimulation of the cardiac branch of the vagus nerve evoked synaptic potentials (spikes and/or excitatory postsynaptic potentials) with an onset latency between 4 and 220 ms. Some neurons displayed both short and long latency inputs(15.5 +/- 1.8 and 160.0 +/- 8.5 ms; n = 14). Of these 78 neurons, 24 responded to veratridine stimulation of cardiac receptors (i.e., cardioreceptive neurons) by exhibiting an augmenting-decrementing discharge of 37 +/- 4 s in duration with a peak frequency of 30 +/- 5 Hz. Convergence from other cardiorespiratory receptors was noted involving either carotid chemoreceptors (n = 7) or pulmonary C-fibers (n = 4) or from both carotid chemoreceptors and pulmonary C-fibers (n = 6). In contrast, only one cardioreceptive NTS neuron was activated by distension of the carotid sinus. Recording sites recovered were confined to the medial NTS at the level of the area postrema and extended caudally into the commissural subnucleus. Our results indicate a convergence of carotid chemoreceptor and pulmonary C-fiber afferent inputs to cardioreceptive NTS neurons. With the paucity of baroreceptor inputs to these neurons it is suggested that sensory integration within the NTS may reflect regulatory versus defensive or protective reflex control.

Afferent Pathways↗
PubMed1997

Hypoxic response of hypoglossal motoneurones in the in vivo cat.

1. In current and voltage clamp, the effects of hypoxia were studied on resting and synaptic properties of hypoglossal motoneurones in barbiturate-anaesthetized adult cats. 2. Twenty-nine hypoglossal motoneurones with a mean membrane potential of -55 mV responded rapidly to acute hypoxia with a persistent membrane depolarization of about +17 mV. This depolarization correlated with the development of a persistent inward current of 0.3 nA at holding potentials close to resting membrane potential. 3. Superior laryngeal nerve (SLN) stimulation-evoked EPSPs were reduced in amplitude by, on average, 46% while IPSP amplitude was reduced by 31% SLN stimulation-evoked EPSCs were reduced by 50-70%. 4. Extracellular application of adenosine (10 mM) hyperpolarized hypoglossal motoneurones by, on average, 5.6 mV, from a control value of -62 mV. SLN stimulation-evoked EPSPs decreased by 18% and IPSPs decreased by 46% during adenosine application. 5. Extracellular application of the KATP channel blocker glibenclamide led to a blockade of a persistent outward current and a significant increase of SLN stimulation-evoked EPSCs. 6. We conclude that hypoglossal motoneurones have a very low tolerance to hypoxia. They appear to be under metabolic stress even in normoxia and their capacity to activate protective potassium currents is limited when compared with other brainstem neurones. This may help to explain the rapid disturbance of hypoglossal function during energy depletion.

Adenosine↗
PubMed1997

Demonstration of glutamate immunoreactivity in vagal sensory afferents in the nucleus tractus solitarius of the rat.

To investigate whether glutamate is a neurotransmitter in vagus nerve sensory afferents terminating in the nucleus tractus solitarius, these terminals were identified by the anterograde transport and their glutamate content examined using the post-embedding immunogold technique. After injection of horseradish peroxidase into the nodose ganglion anterogradely labelled axonal boutons were visualized throughout the nucleus of the solitary tract (nTS), the dorsal motonucleus of the vagus nerve (DVN), predominantly ipsilateral to the injection, and to a lesser extent in the area postrema. Electron microscopic analysis of 47 anterogradely labelled boutons in the nTS following post-embedding immunocytochemistry for glutamate revealed that 43 of these boutons (> 91%) contained a level of glutamate immunoreactivity significantly greater (P < 0.001%) than that observed in the surrounding tissue. The observed enrichment of glutamate immunoreactivity in boutons identified as vagus nerve sensory afferents indicate that glutamate may be a transmitter in these neurones.

Animals↗
PubMed1997

A source of adenosine involved in cardiovascular responses to defense area stimulation.

We have investigated the source of central adenosine important in modulating the cardiovascular response to hypothalamic defense area (HDA) stimulation in alpha-chloralose-anesthetized rats. Microinjections of an ecto-5'-nucleotidase inhibitor, alpha,beta-methylene ADP (alpha,beta-meADP), were made into caudal nucleus of the solitary tract (cNTS) and rostral ventrolateral medulla (RVLM), and its effects on HDA-evoked responses were observed. Stimulation of HDA evoked an increase in arterial pressure and a secondary rise in arterial pressure after stimulation ceased. There was also an increase in heart rate and hindlimb blood flow. alpha,beta-meADP had no effect on resting levels of arterial pressure, heart rate, and hindlimb blood flow when injected into the cNTS or RVLM. alpha,beta-meADP also had no effect on the HDA-evoked tachycardia and increase in muscle blood flow. However, alpha,beta-meADP reduced the primary increase in arterial pressure evoked by HDA stimulation when microinjected into the cNTS. In contrast, alpha,beta-meADP reduced the secondary increase in arterial pressure when microinjected into the RVLM. From these results, we suggest that at least part of the adenosine released centrally during HDA stimulation is derived extracellularly from ATP metabolism.

5'-Nucleotidase↗
PubMed1997

Stimulation within the rostral ventrolateral medulla can evoke monosynaptic GABAergic IPSPs in sympathetic preganglionic neurons in vitro.

The inhibitory responses of identified sympathetic preganglionic neurons (SPNs) to stimulation within the rostral ventrolateral medulla (RVLM) were studied to determine their nature and pharmacology. Whole cell patch-clamp recordings were made from 36 SPNs in the upper thoracic segments of the spinal cord in a neonatal rat brain stem-spinal cord preparation. Neurons were identified as SPNs on the basis of their antidromic activation after stimulation of the ipsilateral segmental ventral root and their morphology and location in the intermediolateral cell column and intercalated nucleus. In all SPNs, electrical stimulation of the RVLM evoked fast excitatory postsynaptic potentials (EPSPs) that were mediated by non-N-methyl-D-aspartate (NMDA) and NMDA receptors. These excitatory responses were the most prominent response in control artificial cerebrospinal fluid and have been studied previously. In 22 of the SPNs, RVLM stimulation also elicited fast inhibitory postsynaptic potentials (IPSPs), which increased in amplitude as the membrane was depolarized. Five of these neurons were not studied further as they responded occasionally with IPSPs that had highly variable onset latencies indicating the involvement of a polysynaptic pathway. In the remaining SPNs (n = 17), the evoked IPSPs persisted in the presence of the excitatory amino acid antagonists 6-cyano-7-nitroquinoxaline-2,3,-dione and D,L-2-amino-5-phosphonopentanoic acid. In eight of these SPNs, it was necessary to block the EPSPs to reveal the IPSPs. In the 7 SPNs tested, the onset latencies of the IPSPs were not significantly different from the onset latencies of the fast EPSPs. The low sweep-to-sweep fluctuations in onset latency of individual IPSPs (absolute average deviation: 0.4 ms) indicated that the IPSPs were elicited by activation of a monosynaptic pathway. The amplitudes of the IPSPs decreased in amplitude as the membrane was hyperpolarized and reversed in polarity at -70.3 +/- 1.7 mV (mean +/- SD), which was close to the equilibrium potential for chloride ions. In addition, in seven SPNs, bath applications of 5 microM bicuculline, a gamma-aminobuturic acid-A (GABAA) antagonist, abolished or reduced the evoked IPSPs. Five SPNs also were studied that displayed ongoing IPSPs. The amplitudes of these IPSPs increased with membrane depolarization and were blocked by bath applications of 5 microM bicuculline, suggesting that they also were mediated by activation of GABAA receptors. These results demonstrate the existence of a bulbospinal GABAergic pathway impinging directly onto SPNs. This pathway may be tonically active in the neonatal rat brain stem-spinal cord preparation.

Adrenergic Fibers↗