Excitatory and indirect inhibitory actions of 5-hydroxytryptamine on sympathetic preganglionic neurones in the neonate rat spinal cord in vitro

The action of 5-hydroxytryptamine (5-HT) on sympathetic preganglionic neurones (SPN) was studied by intracellular recordings in thin slices of neonatal rat spinal cord in vitro. Superfusion of 5-HT (1–270 μM) to SPN caused a concentration dependent slow depolarization or inward current and an increase in synaptic activity consisting of both EPSPs and IPSPs. The slow depolarization was still present after superfusion with TTX. Similar effects were seen during superfusion with 5-carboxamidotryptamine (5-CT) or -methyl-5-hydroxytryptamine (-me-5-HT). A comparison with the potency of 5-HT was made for 5-CT or -me-5-HT on the same neurone by determining the magnitude of the slow depolarization to different concentrations of agonist. This showed that the apparent potency of the agonists was 5-CT> 5-HT> -me-5-HT even in the presence of fluoxetine, a 5-HT uptake inhibitor. The 5-HT-induced slow depolarization was partially blocked by ketanserin but full recovery was not observed. The results suggest that the excitatory action of 5-HT on SPN is mediated via an atypical 5-HT₂ receptor or a 5-HT_1C-like receptor. The 5-HT-induced IPSPs were reversibly blocked by superfusion with strychnine, suggesting they were mediated by glycine.     

A comparison of the effects of intrathecally administered 5-hydroxytryptamine and thyrotropin-releasing hormone on renal and muscle sympathetic nerve activity

Thyrotropin-releasing hormone (TRH) has been shown to coexist with 5-hydroxytryptamine (5-HT) in spinally projecting raphe neurones, some of which terminate in the sympathetic nuclei of the spinal cord. In an attempt to mimic the actions of these neurones, the effects of intrathecal administration of 5-HT was compared to that of TRH on activity in two sympathetic postganglionic nerves, the renal nerve and sympathetic fibres to skeletal muscle of the hind limb. In chloralose-urethane anaesthetised rats intrathecal infusion of TRH at the T9 level in doses of 5-30 micrograms increased activity in renal nerve as did low doses (20-100 micrograms) of 5-HT. In contrast, intrathecal infusion of TRH in doses of 5-30 micrograms and 5-HT in doses of 20-800 micrograms at L4 level decreased activity in sympathetic fibres to muscle. The results show that TRH, unlike 5-HT, has an excitatory but not an inhibitory action on renal sympathetic activity whereas both substances have only an inhibitory action on muscle sympathetic activity.     

Excitatory and inhibitory effects of epinephrine on neonatal rat sympathetic preganglionic neurons in vitro

Current and voltage recordings were made from antidromically identified sympathetic preganglionic neurons (SPNs) in transverse thoracolumbar spinal cord slices removed from neonatal rats. When applied by either pressure ejection or superfusion, epinephrine (Epi) caused a slow depolarization or an inward current in 62 SPNs (42%) and a slow hyperpolarization or an outward current in 21 SPNs (14%). The responses persisted in low calcium- or tetrodotoxin-containing media. The Epi-induced depolarization or inward current was associated with increased membrane resistance; it was reduced by membrane hyperpolarization and nullified at a membrane potential of about -100 mV; a clear reversal however was not observed at more negative potential levels. In a number of SPNs the Epi-induced depolarization was accompanied by small inhibitory postsynaptic potentials. The latter were eliminated by a low calcium solution and by the glycine antagonist strychnine, suggesting that they were caused by glycine or a glycine-like substance released from interneurons subsequent to activation by Epi. The Epi-induced hyperpolarization or outward current was associated with decreased membrane resistance, and nullified around -100 mV. The alpha-adrenergic antagonist, dihydroergotamine, and alpha 1-antagonist, prazosin, reversibly blocked the excitatory, whereas the alpha 2-antagonist, yohimbine, abolished the inhibitory response, respectively. It is concluded that Epi acting on alpha 1- and alpha 2-adrenergic receptors depolarizes and hyperpolarizes the rat SPNs by decreasing or increasing membrane conductances to potassium ions.     

Effect of intrathecally administered noradrenergic antagonists on nociception in the rat

Recent evidence suggests that some groups of noradrenergic neurons found in the brainstem have axonal connections in the spinal cord dorsal horn and may be involved in the control of pain sensitivity. Such evidence includes the demonstration that intrathecal injection of noradrenergic agonists increases nociceptive threshold (hypoalgesia). The present studies examined whether the descending noradrenergic system is tonically active and, if so, what noradrenergic receptor subtypes mediate the actions of endogenously-released norepinephrine. These studies involved the measurement of nociceptive threshold before and after the intrathecal injection of noradrenergic antagonists having different relative affinities for α-noradrenergic receptor subtypes.The intrathecal administration of α-noradrenergic antagonists produced a dose-dependent decrease in nociceptive threshold (hyperalgesia). This finding is consistent with the proposal that tonically-active bulbospinal noradrenergic neurons modulate the processing of nociceptive information in the spinal cord. The potency and duration of the hyperalgesia was correlated with the relative potency of the antagonists for the α-2 noradrenergic receptor. The relative potencies were as follows: yohimbine > phentolamine > WB 4101 > prazosin. Thus, endogenous norepinephrine which is tonically released from bulbospinal axon terminals may interact preferentially with noradrenergic receptors of the α-2 type.     

Intrathecally administered angiotensin II increases sympathetic activity in the rat

The possibility that angiotensin II (AII) functions as an excitatory transmitter on sympathetic preganglionic neurones was tested in anaesthetized rats. Drugs were administered intrathecally whilst recording blood pressure, heart rate and sympathetic activity in splanchnic or renal nerves. Intrathecal AII (20 microliters, 10(-5) M) caused a significant increase in blood pressure of 13% +/- 3 and in sympathetic activity of 15% +/- 5. Intrathecal AII (20 microliters, 10(-3) M) caused larger increases in blood pressure of 22% +/- 3 and in sympathetic activity of 25% +/- 3. The magnitude of the response was dependent on the location of the catheter tip within the subarachnoid space T9-T11 being best for the above changes. Preceding intrathecal AII with the AII antagonist Saralasin (20 microliters, 10(-3) M) also given intrathecally prevented the changes. An increase in sympathetic nerve activity brought about reflexly by a drop in blood pressure of 45-47 mm Hg was almost halved by intrathecal Saralasin. Peak plasma counts (less than 5% of total) of [3H]AII in the blood after intrathecal injection occurred 2 min after the peak changes in sympathetic activity and blood pressure. Counts of [3H]AII in the spinal cord showed that 81% of recovered label was within one segment on either side of the catheter tip. It is concluded that AII has an excitatory action on sympathetic neurones in the spinal cord.     

Intrathecal 5-hydroxytryptamine and electrical stimulation of the nucleus raphe magnus in rats both reduce the antinociceptive potency of intrathecally administered noradrenaline

The antinociceptive potency of noradrenaline (NA), as assessed by suppression of the spinal nocifensive tail flick and paw withdrawal reflexes was examined. The tail flick latency (TFL) was increased to the cut-off value for a period of approximately 120 min following the intrathecal microinjection of a standard 15 nmol dose of NA. A similar intrathecal dose of NA produced an increase in threshold to elicit the paw withdrawal reflex for a period of approximately 20 min. When preceded by a standard intrathecal microinjection of 260 nmol 5-hydroxytryptamine, the antinociceptive potency of NA was significantly reduced as reflected by both the tail flick and paw withdrawal tests. In addition, electrical stimulation of the posterior raphe complex immediately before and during the NA-induced increase in TFL, significantly reduced the antinociceptive potency of NA. It is concluded that spinal tryptaminergic activity can reduced the duration of the antinociceptive action of intrathecally applied NA.     

Excitatory and inhibitory effects of lingual nerve stimulation on reflexes controlling the activity of masseteric motoneurons

The masseteric reflex was suppressed for 200–400 msec following a single shock stimulus to low threshold fibers in either the ipsilateral or contralateral lingual nerve of decerebrate cats. There were two phases of suppression; the maximum reduction in the amplitude of the reflex occurred 5–10 msec after the stimulus in the first phase, and approximately 40 msec after the stimulus in the second phase. Between these two phases of suppression was a recovery period characterized by an increase in reflex amplitude.     

Excitatory and inhibitory actions of ibotenic acid on frog spinal motoneurones in vitro

Ibotenic acid (IBO), a conformationally-restricted analogue of the putative excitatory transmitter glutamate, produced biphasic effects on frog spinal motoneurones recorded with DC-coupled Ag/AgCl electrodes placed on the central end of ventral roots. The initial effect consisted of a motoneuronal depolarization with an increase in excitability tested with low-intensity dorsal root stimulation (the depolarizing activity of IBO was about 6 times greater than that of glutamate). On washout, the depolarizing effect of IBO subsided and complete motoneuronal repolarization occured; however, motoneuronal responses to glutamate and to trans-synaptic stimulation remained depressed for prolonged periods. This depressant effect was not reproduced following prolonged administration of depolarizing doses of glutamate but it was similar to that produced by muscimol, an agonist on inhibitory GABA receptors. Unlike the muscimol-evoked depression, the IBO-induced depression was insensitive to bicuculline and was antagonized by local anaesthetics or high Mg²⁺ solutions. It is suggested that a substantial part of the IBO depressant action is likely due to release of long-acting inhibitory transmitters onto motoneurones.     

Effects of activating spinal alpha-adrenoreceptors on sympathetic nerve activity in the rat

The effects of several alpha-adrenoreceptor agonists and antagonists administered intrathecally at T10 level on renal sympathetic nerve activity (RSNA) were examined, in chloralose-urethane anaesthetised rats. Intrathecal noradrenaline (NA, 0.5-500 micrograms) produced one of 3 responses depending on dose, an inhibition of RSNA at low doses, an excitation of RSNA at high doses, or a biphasic effect. Intrathecal adrenaline (5-200 micrograms) was inhibitory in the main but some doses elicited poorly repeatable brief excitation followed by prolonged inhibition. Intrathecal methoxamine (ME; 2.5 ng-25 micrograms) caused a dose-dependent increase in RSNA (mean maximum response 27 +/- 0.5%). The excitatory effects of NA and ME were blocked (72% +/- 12%) by pretreatment with intrathecal prazosin (PRA, 20-200 ng) but not by yohimbine (YOH, 200 ng). Intrathecal guanabenz (GUA 3-15 micrograms) caused a dose-dependent inhibitory effect on RSNA (mean maximum 32% +/- 5%). The inhibitory effects of NA, adrenaline and GUA were blocked by pretreatment with intrathecal YOH (200 ng-2 micrograms). Intrathecal PRA (200 ng) had no effect on the inhibitory effects of NA and GUA. Intravenous administration of each of the adrenoreceptor agonists (apart from adrenaline), at similar doses to those given intrathecally, in most cases had no significant effect on RSNA; in a few cases the opposite effects to those produced by intrathecal administration were seen.     

Depression of C fiber-evoked activity by intrathecally administered reactive red 2 in rat thalamic neurons

To investigate the possible role of spinal purinoceptors in nociception, the potent P₂-purinoceptor antagonist reactive red 2 was studied in rats under urethane anesthesia in which nociceptive activity was elicited by electrical stimulation of afferent C fibers in the sural nerve and recorded from single neurons in the ventrobasal complex of the thalamus. Intrathecal (i.t.) application of reactive red 2 (6–200 μg) caused a dose-dependent reduction of the evoked activity in thalamic neurons. The estimated ED₅₀ was 30 μg, and the maximum depression of nociceptive activity amounted to about 70% of the control activity at a dose of 100 μg. Morphine, administered i.t. at a maximally effective dose (80 μg), inhibited the evoked nociceptive activity by only up to 55% of the control activity. An i.t. co-injection of reactive red 2 (100 μg) and morphine (80 μg) caused a maximum reduction of the evoked thalamic activity by up to 85% of the control activity, thus, exceeding significantly the effect elicited by either drug alone. Similarly, i.t. co-injection of almost equipotent dosages of reactive red 2 (30 μg) and morphine (30 μg) caused a maximum reduction of the evoked activity by up to 72% of the control activity, which again exceeded significantly the effect of either drug alone. The results suggest that in rats reactive red 2 exerts antinociception by blockade of P₂-purinoceptors in the spinal cord and, hence, support the idea that ATP may play an important role in spinal transmission of nociceptive signals. An activation of the spinal opioid system does not seem to contribute to the effect of reactive red 2 but might act additive or even synergistically with its antinociceptive action.     

鞘内注射催产素对大鼠神经痛反应的影响

目的 :观察不同剂量催产素对神经痛大鼠热痛敏的影响。方法 :在脊神经结扎致坐骨神经损伤大鼠模型上采用辐射热缩腿反射的方法 ,以抬脚潜伏期作为观察指标。结果 :蛛网膜下腔注射催产素 (1ng ,2 5ng ,5ng)对神经痛大鼠有镇痛作用 ,呈剂量相关关系。结论 :鞘内注射催产素对神经痛大鼠有镇痛作用。     

Chemical stimulation of the locus coeruleus: inhibitory effects on hemodynamics and renal sympathetic nerve activity

We examined the role of the locus coeruleus (LC) in the regulation of the hemodynamics and sympathetic nerve activity in anesthetized rats. Unilateral microinjection into the LC of the excitatory amino acid, L-glutamate (Glu), elicited dose-dependent decreases in arterial pressure (AP) and heart rate (HR). The bradycardic response was partially attenuated after intravenous injection of atropine sulfate, but the greater part of this response still remained. Interruption of the ascending projections of the LC by midbrain transection did not affect the depressor and bradycardic responses elicited by chemical stimulation. The renal sympathetic nerve activity showed transient but strong inhibition with this stimulation. Cardiac output was measured using an electromagnetic flowmeter implanted in the ascending aorta. The stroke volume and total peripheral resistance (TPR) were calculated. Microinjection of Glu elicited a significant decrease in TPR and slight decreases in cardiac output and stroke volume. Microinjection of the inhibitory amino acid, gamma-aminobutyric acid (GABA), or the alpha 2-adrenergic agonist, clonidine, exerted no effect on AP and HR. The present results therefore suggest that: (1) the LC neurons have an inhibitory influence on the sympathetic nervous system, and stimulation of these neurons can elicit depressor and bradycardic responses; (2) the depressor response was produced predominantly as a result of a decrease in vascular resistance, rather than a decrease in cardiac output; (3) these inhibitory responses may be provided not via the ascending projections of the LC; and (4) the LC neurons do not have a tonic influence on the cardiovascular system.     

Effects of centrally administered angiotensin on sympathetic nerve activity and blood flow to the kidney in conscious rats

The effects of intracerebroventricular (i.c.v.) administration of different doses (10 pg-100 ng) of angiotensin II (AII) on renal sympathetic nerve activity (RSNA), mean arterial blood pressure (MAP), heart rate (HR), renal blood flow and femoral blood flow have been examined in conscious rats. Administration of AII (10 ng) through a chronically implanted cannula induced an increase in MAP (20-22 mmHg), a decrease in HR (24 bpm), a decrease in RSNA by 57%, a decrease of femoral blood flow by 21% but no change in renal blood flow. The effects on MAP, HR and RSNA are greatly attenuated by the prior i.c.v. injection of an AII-antagonist saralasin. In anesthetized rats, renal denervation significantly attenuated an increase in urinary sodium excretion induced by i.c.v. injection of AII. Since activation of the renal nerve is known to induce sodium reabsorption from the renal tubule and renin release, the relevance of the present finding is discussed in relation to the effect of AII on sodium excretion.     

Excitatory and inhibitory influences on bladder activity elicited by electrical stimulation in the pontine micturition center in the rat

Electrical stimulation at various sites in the dorsal pontine tegmentum in urethane anesthetized rats modulated activity of the urinary bladder as well as efferent firing on bladder postganglionic nerves. Electrical stimulation (0.2 ms 50 Hz, 5-20 V or 30-150 microA, 2-5 s train duration) using a microelectrode (tip diameter, 10-20 microns) in an excitatory area located rostral and medial to the locus coeruleus evoked short latency (less than 2 s) large amplitude (greater than 20 cm H2O) bladder contractions and increased firing on the bladder postganglionic nerves. Stimulation at sites adjacent to the excitatory area inhibited bladder postganglionic nerve firing and bladder activity. Inhibitory responses were evident as either a decrease in intravesical pressure, an increased interval between bladder contractions, or an interruption or elimination of bladder contractions. The threshold intensity for excitation using a large electrode (2-4 V) was slightly higher than that for inhibition (1.5-2 V). The optimum sites for evoking bladder contractions were located in and close to the laterodorsal tegmental nucleus (LDT) and in the periaqueductal gray just dorsal or dorsolateral to the LDT. The extent of the area that induced bladder contractions was 0.5-1.2 mm in diameter in each rat when a microelectrode was employed for electrical stimulation. Electrical stimulation in the optimum site for evoking bladder contractions induced relatively little striated muscle activity and produced no short-latency blood pressure changes. The longer latency blood pressure changes associated with a spontaneous bladder contraction were still present following a stimulation of the dorsolateral pons. These data are consistent with the view that neurons in the dorsal pontine tegmentum play an important role in the regulation of urine storage as well as urine release.