Cardiac nociceptors and ischemia: role of sympathetic afferents in cat

In total, 86 units were recorded from T2 and T3 left thoracic rami of cat. These receptors were located on the circumflex coronary artery, anterior descending coronary artery, and its adjacent myocardial regions. The conduction velocity of these fibres was in the range of "C" (0.5 to 1.8 m/s) and "A delta" (5.96 to 17 m/s) fibres. Out of these 86 units, only 28 units were activated by coronary occlusion. The average resting frequency of the spontaneous unit was 0.8 +/- 0.06 impulses/s which increased to 35 +/- 4.8 impulses/s on mechanical probing. In order to examine whether the units sensitive to coronary occlusion were also responsive to algesic agents, some of these units were studied applying lactic acid, bradykinin, prostaglandins, and nicotine. It was observed that these ischemia-sensitive units are also sensitive to lactic acid (10 units), bradykinin (16 units), prostaglandins (12 units), and nicotine (15 units). These ischemia-sensitive units are presumably nociceptors and activated by algestic agents that cause cardiac ischemic pain.     

20-HETE increases renal sympathetic nerve activity via activation of chemically and mechanically sensitive muscle afferents

Arachidonic acid and its metabolites produced via cyclooxygenase (COX) and lipoxygenase pathways have been reported to contribute to the cardiovascular reflexes evoked by stimulating thin fibre muscle afferents during muscle contraction. 20-Hydroxyeicosatetraenoic acid (20-HETE), a primarily metabolized product of arachidonic acid by cytochrome P450 enzymes, can be accumulated in contracting muscles. Thus, the purpose of this study was to determine the role of 20-HETE in modulating the reflex sympathetic responses to activation of chemically and mechanically sensitive muscle afferents. The renal sympathetic nerve activity (RSNA) and cardiovascular responses were examined after injections of 20-HETE into the arterial blood supply of the hindlimb muscles of decerebrated rats. This induced a dose-dependent increases in RSNA and mean arterial pressure (MAP). We also tested the hypothesis that 20-HETE would sensitize muscle afferents and, thereby, augment the RSNA and blood pressure response to muscle stretch. The results show that arterial infusion of 20-HETE significantly enhanced the RSNA and MAP responses to muscle stretch. In contrast, N -hydroxy- N '-(4-butyl-2-methylphenyl)formamidine, a potent inhibitor of 20-HETE production, attenuated the reflex muscle responses. Furthermore, the sensitizing effect of 20-HETE on the muscle reflex was significantly attenuated after blocking COX activity with indomethacin. Our data suggest that 20-HETE plays a role in modulating muscle afferent-mediated sympathetic responses, probably through engagement of a COX-dependent mechanism.     

Tyramine-induced endogenous noradrenaline efflux from in situ cardiac sympathetic nerve ending in cats

With the use of dialysis technique, the effects of tyramine on in situ cardiac sympathetic nerve endings were examined in anaesthetized cats. Dialysis probes were implanted in the left ventricular myocardium, and the concentration of dialysate noradrenaline (NA) served as an indicator of NA output at the cardiac sympathetic nerve ending. Locally applied tyramine (600 microM) increased dialysate NA levels from 17 +/- 1 (pg mL-1) to 3466 +/- 209 (pg mL-1). Pretreatment with reserpine (vesicle transport NA blocker 1 microM) did not affect tyramine-induced NA efflux. The tyramine-induced NA efflux was augmented by pretreatment with pargyline (1 mM) but suppressed by pargyline (10 mM). Pretreatment with alpha-methyl-tyrosine suppressed NA efflux evoked by tyramine. These pretreatments did not affect the time course of NA efflux but only altered peak height of NA efflux. The efflux of NA evoked by tyramine was not associated with any reduction of dihydroxyphenylglycol (DHPG). In contrast, in the pretreatment with reserpine, the efflux of NA was associated with a reduction of DHPG. This result suggests that NA graduation between axoplasm and stored vesicle contributes to maintaining the axoplasmic NA level during carrier-mediated outward NA transport. The tyramine-induced NA efflux provides a close reflection of the NA content at the nerve ending. With the use of dialysis, this experimental model is suitable for studying the mechanism of sympathomimetic amine-induced neurotransmitter efflux.     

Responses of medullary raphespinal neurons to electrical stimulation of thoracic sympathetic afferents, vagal afferents, and to other sensory inputs in cats.

1. Medullary raphespinal neurons antidromically activated from the T2-T5 segments were tested for responses to electrical stimulation of cervical vagal and thoracic sympathetic afferents (by stimulating the left stellate ganglion), somatic probing, auditory stimuli, and visual stimuli in cats anesthetized with alpha-chloralose. A total of 99 neurons in the raphe nuclei were studied; the locations of 76 cells were histologically confirmed. Neurons were located in raphe magnus (RM, 65%), raphe obscurus (RO, 32%), and raphe pallidus (RPa, 4%). The mean conduction velocity of these neurons was 62 +/- 2.9 (SE) m/s with a range of 1.1-121 m/s. 2. A total of 60/99 tested neurons responded to electrical stimulation of sympathetic afferents. Quantitation of responses was obtained for 55 neurons. With one exception, all responsive neurons were excited and exhibited an early burst of spikes with a mean latency of 16 +/- 1.2 ms. From a spontaneous discharge rate of 5.2 +/- 1.2 spikes/s, neuronal activity increased by 2.9 +/- 0.3 spikes/stimulus. In addition to an early peak, 15 neurons (25%) exhibited a late burst of spikes with a latency of 182 +/- 12.9 ms; neuronal activity increased by 5.0 +/- 1.3 spikes/stimulus. Duration of the late peak (130 +/- 18.5 ms) was longer than for the early peak (18 +/- 0.7 ms), but threshold voltages for eliciting each peak were comparable. Sixteen of 29 spontaneously active neurons exhibited a postexcitatory depression of activity that lasted for 163 +/- 19.1 ms. All but one tested neuron in RO responded to stimulation of sympathetic afferents, but 65% of neurons in RM responded to this stimulus. 3. In response to vagal afferent stimulation, 19% of 57 neurons exhibited inhibition only, 11% were only excited, and 9% were either excited or inhibited, depending on the stimulus paradigm used; the remaining 61% of neurons were unresponsive. From a spontaneous rate of 7.9 +/- 3.8 spikes/s, excited cells increased their discharge rate by 1.6 +/- 0.3 spikes/stimulus. Activity of inhibited cells was reduced from 21.3 +/- 5.8 to 7.8 +/- 3.1 spikes/s. The conditioning-test (CT) technique was used to assess 11 neurons' responses. Stellate ganglion stimulation was the test stimulus, and vagal stimulation the conditioning stimulus. Vagal stimulation reduced the neuronal responses to stellate ganglion stimulation by an average of 50% with a CT interval of 60-100 ms, and cell responses returned to control after 300 ms. With spontaneous cell activity, low frequencies of vagal stimulation were generally excitatory, and high frequencies (10-20 Hz) inhibitory.(ABSTRACT TRUNCATED AT 400 WORDS)     

Types of afferents from the knee joint evoking sympathetic reflexes in cat inferior cardiac nerves

In anesthetized cats electrical stimulation of the medial articular nerve of the knee joint evoked sympathetic reflex discharges in inferior cardiac nerves. Low intensity single stimuli elicited early reflex discharges (A-reflexes, latency 70-90 ms, duration 110-200 ms) whereas short tetanic stimulation at higher intensities evoked, in addition, late reflexes (C-reflexes, latency 390-480 ms, duration 230-400 ms). An analysis of the relation between the conduction velocity and the electrical threshold of 231 single medial articular nerve fibers revealed that the A-reflex is mainly due to activation of Group II units, whereas the C-reflex is evoked by activity in unmyelinated Group IV fibers.     

Undiscovered role of endogenous thromboxane A2 in activation of cardiac sympathetic afferents during ischaemia

Myocardial ischaemia activates blood platelets, which in turn stimulate cardiac sympathetic afferents, leading to chest pain and sympathoexcitatory reflex cardiovascular responses. Previous studies have shown that activated platelets stimulate ischaemically sensitive cardiac sympathetic afferents, and that thromboxane A(2) (TxA(2)) is one of the mediators released from activated platelets during myocardial ischaemia. The present study tested the hypothesis that endogenous TxA(2) stimulates cardiac afferents during ischaemia through direct activation of TxA(2) (TP) receptors coupled with the phospholipase C-protein kinase C (PLC-PKC) cellular pathway. Nerve activity of single unit cardiac sympathetic afferents was recorded from the left sympathetic chain or rami communicantes (T(2)-T(5)) in anaesthetized cats. Single fields of 39 afferents (conduction velocity = 0.27-3.65 m s(-1)) were identified in the left or right ventricle initially with mechanical stimulation and confirmed with a stimulating electrode. Five minutes of myocardial ischaemia stimulated all 39 cardiac afferents (8 Adelta-, 31 C-fibres) and the responses of these 39 afferents to chemical stimuli were further studied in the following four protocols. In the first protocol, 2.5, 5 and 10 microg of the TxA(2) mimetic, U46619, injected into the left atrium (LA), stimulated seven ischaemically sensitive cardiac afferents in a dose-dependent manner. Second, BM13,177, a selective TxA(2) receptor antagonist, abolished the responses of six afferents to 5 microg of U46619 injected into the left atrium and attenuated the ischaemia-related increase in activity of seven other afferents by 44%. In contrast, cardiac afferents, in the absence of TP receptor blockade responded consistently to repeated administration of U46619 (n = 6) and to recurrent myocardial ischaemia (n = 7). In the fourth protocol, administration of PKC-(19-36), a selective PKC inhibitor, attenuated the responses of six other cardiac afferents to U46619 by 38%. Finally, using an immunohistochemical staining approach, we observed that TP receptors were expressed in cardiac sensory neurons in thoracic dorsal root ganglia. Taken together, these data indicate that endogenous TxA(2) contributes to the activation of cardiac afferents during myocardial ischaemia through direct stimulation of TP receptors probably located in the cardiac sensory nervous system and that the stimulating effect of TxA(2) on cardiac afferents is dependent, at least in part, upon the PLC-PKC cellular pathway.     

Myocardial nerve fibers are preserved in MPTP-treated mice, despite cardiac sympathetic dysfunction

Recently, we reported a profound depletion of cardiac sympathetic nerve fibers in Parkinson's disease (PD). This cardiac sympathetic denervation is a characteristic hallmark of PD. Cardiac sympathetic dysfunction was also observed in 1-methyl-4-phenyl-1,2,3,6-tetrahydroxypyridine (MPTP)-treated mice, a model of PD. Although binding assay showed a decreased density of norepinephrine transporter (NET) in the hearts of the mice, their histopathological alterations have not been demonstrated. In this study, we investigated hearts of MPTP-treated mice with immunohistochemical method and Western blot analyses. MPTP-treated mice showed significant decreases in the contents of cardiac noradrenaline and dopamine, suggesting the sympathetic dysfunction. Synaptophysin-, tyrosine hydroxylase- or NET-immunoreactive nerve fibers were abundant in the hearts of control mice and MPTP-treated mice, without apparent differences between the two groups. Western blot analyses also showed no difference in the amounts of these proteins. Myocardial nerve fibers were well preserved in MPTP-treated mice, despite apparent cardiac sympathetic dysfunction.     

Tyramine‐induced endogenous noradrenaline efflux from in situ cardiac sympathetic nerve ending in cats

With the use of dialysis technique, the effects of tyramine on in situ cardiac sympathetic nerve endings were examined in anaesthetized cats. Dialysis probes were implanted in the left ventricular myocardium, and the concentration of dialysate noradrenaline (NA) served as an indicator of NA output at the cardiac sympathetic nerve ending. Locally applied tyramine (600 μM ) increased dialysate NA levels from 17 ± 1 (pg mL^?1) to 3466 ± 209 (pg mL^?1). Pretreatment with reserpine (vesicle transport NA blocker 1 μM ) did not affect tyramine‐induced NA efflux. The tyramine‐induced NA efflux was augmented by pretreatment with pargyline (1 m M ) but suppressed by pargyline (10 m M ). Pretreatment with α‐methyl‐tyrosine suppressed NA efflux evoked by tyramine. These pretreatments did not affect the time course of NA efflux but only altered peak height of NA efflux. The efflux of NA evoked by tyramine was not associated with any reduction of dihydroxyphenylglycol (DHPG). In contrast, in the pretreatment with reserpine, the efflux of NA was associated with a reduction of DHPG. This result suggests that NA graduation between axoplasm and stored vesicle contributes to maintaining the axoplasmic NA level during carrier‐mediated outward NA transport. The tyramine‐induced NA efflux provides a close reflection of the NA content at the nerve ending. With the use of dialysis, this experimental model is suitable for studying the mechanism of sympathomimetic amine‐induced neurotransmitter efflux.     

Modulatory effects of ketamine on catecholamine efflux from in vivo cardiac sympathetic nerve endings in cats

With the use of the microdialysis technique, we examined the modulatory effect of ketamine on catecholamine efflux from in vivo cardiac sympathetic nerve endings. A dialysis probe was implanted in the left ventricular myocardium, and dialysate norepinephrine (NE) levels in anesthetized cats were measured with liquid chromatogram-electrical detection. A 60-min occlusion of the left anterior descending coronary artery caused increases in dialysate NE levels. Through the dialysis probe, locally applied ketamine (10 mM) augmented the dialysate NE responses to coronary occlusion in the presence and absence of desipramine (membrane NE transport blocker). Thus, the ketamine-induced NE increment is not mediated through the neuronal NE transporter. The sympathomimetic action of ketamine may augment the NE efflux evoked by myocardial ischemia.     

High threshold aortic baroreceptor afferents in the sympathetic nerve

In anaesthetized cats, 40 sympathetic sensory units in the bracheocephalic artery (30 units) and the descending aorta (10 units) were recorded by means of single-unit preparation. Direct evidence is available for the mechanosensitive nature of the receptors in the sympathetic afferents at the level of T3 and T4. Two distinct types of receptors were found (Type I and Type II). Type I receptors, which were fast adapting, gave a spike discharge at each systolic height of pressure (70-110 mmHg). However, they sometimes failed to appear even at such systolic pressure. When the systemic pressure was increased by occluding the descending aorta or by infusing adrenaline solution intravenously, the frequency of discharge of Type I receptors increased and they behaved much the same as the typical sinoaortic baroreceptors. Type II receptors were activated by mechanical probing and at high systemic pressure, though they did not fire always synchronously with heart beat. On the basis of the study it may be suggested that Type I receptors are high threshold baroreceptors and like other systemic baroreceptors, play a role in homeostatic control presumably in a state of high blood pressure, but on the other hand Type II receptors do not play such a role.     

Fundamental rhythm of cardiac sympathetic nerve activity in awake cats at rest and during body movement

Cardiac sympathetic nerve activity (CSNA in imp/s) was measured in the postganglionic fibers of awake cats at rest, during body movement, and with excitement. The CSNA showed synchronized discharges with various periodicities. Rhythms of the synchronized CSNA were analyzed by an interval histogram (IIH). The IIH showed a multimodal distribution. The first model interval (Tc) was in a range of 75 to 125 ms. An 8-12 cycle/s Tc rhythm, i.e., inverse value of Tc, was always observed in the awake cat at rest and during body movements. Probability of the 8-12 cycle/s Tc rhythm was smallest at rest, increased during body movement, and was largest with excitement. These results suggested that the 8-12 cycle/s Tc rhythm, observed in all states in the conscious cat, is a fundamental rhythm of central cardiovasomotor origin. The subsequent model distribution (Tb = 2 x Tc, 3 x Tc, 4 x Tc, or 5 x Tc) ranged from 150 to 700 ms, mostly 200 to 500 ms. A 2-5 cycle/s Tb rhythm, i.e., inverse value of Tb, appeared more frequently at rest than that during body movement or with excitement. A new model concerning a mechanism to cause the 2-5 cycle/s Tb rhythm is suggested.     

Morphology of single afferents of the saccular macula in cats

The morphology of single saccular afferents was studied by the intracellular horseradish peroxidase (HRP) method. Four neurons were sufficiently stained to allow reconstruction of their axonal arborizations. The main axon of these neurons bifurcated into an ascending and a descending branch at the level of the lateral nucleus. The ascending branches of two axons gave off collaterals with boutons in the caudal part of the superior nucleus, while the other two ascending branches lacked such terminations. By contrast, characteristics of the descending axonal arborization patterns of all the four neurons were substantially the same. The descending branches coursed caudally through the lateral part of the descending nucleus, and gave off up to 14 collaterals with boutons that extended throughout this nucleus. These collaterals also reached the ventral part of the lateral nucleus, the lateral border of the medial nucleus, and group f. A few axon collaterals ramified even outside the border of the vestibular nuclei into the spinal trigeminal nucleus and the reticular formation surrounding it. Axon collaterals from the stem axon also terminated in the interstitial nucleus of the vestibular nerve. There was a noticeable absence of any projection to the y group.