Bevyn Jarrott

Neurochemical modulation of cardiovascular control in the nucleus tractus solitarius.

The central control of cardiovascular function has been keenly studied for a number of decades. Of particular interest are the homeostatic control mechanisms, such as the baroreceptor heart-rate reflex, the chemoreceptor reflex, the Bezold-Jarisch reflex and the Breuer-Hering reflex. These neurally-mediated reflexes share a common termination point for their respective centrally-projecting sensory afferents, namely the nucleus tractus solitarius (NTS). Thus, the NTS clearly plays a critical role in the integration of peripherally initiated sensory information regarding the status of blood pressure, heart rate and respiratory function. Many endogenous neurochemicals, from simple amino acids through biogenic amines to complex peptides have the ability to modulate blood pressure and heart rate at the level of the NTS. This review will attempt to collate the current knowledge regarding the roles of neuromodulators in the NTS, the receptor types involved in mediating observed responses and the degree of importance of such neurochemicals in the tonic regulation of the cardiovascular system. The neural pathway that controls the baroreceptor heart-rate reflex will be the main focus of attention, including discussion of the identity of the neurotransmitter(s) thought to act at baroafferent terminals within the NTS. In addition, this review will provide a timely update on the use of recently developed molecular biological techniques that have been employed in the study of the NTS, complementing more classical research.

Release, spread and persistence of immunoreactive neurokinin A in the dorsal horn of the cat following noxious cutaneous stimulation. Studies with antibody microprobes

In barbiturate anaesthetized spinal cats antibody microprobes were used to examine release of immunoreactive neurokinin A following cutaneous thermal and mechanical stimulation. In the absence of peripheral stimuli, microprobes detected a diffuse basal presence of immunoreactive neurokinin A. Noxious mechanical and to a lesser extent noxious thermal stimuli increased the levels of immunoreactive neurokinin A diffusely throughout the dorsal horn which, in many cases, spread into the adjacent white matter. These diffuse stimulus-evoked increases contrast with previous experiments where the same stimuli produced discrete focal increases in levels of immunoreactive substance P. Evidence was obtained that released immunoreactive neurokinin A persisted in the spinal cord for at least 30 min beyond the period of stimulation. Neurokinin A needs consideration as the agent responsible for the long-lasting increases in excitability of some spinal neurons found by several laboratories to follow a brief input from unmyelinated primary afferents.

Release of immunoreactive substance P in the spinal cord during development of acute arthritis in the knee joint of the cat: a study with antibody microprobes

In anaesthetized spinal cats, the release of immunoreactive substance P in the spinal cord during development of an acute inflammation in one knee joint was studied with antibody microprobes. The microprobes bore antibodies directed to the C- or N-terminus of substance P. With the normal knee joint, innocuous mechanical stimuli (flexion, pressure) did not result in spinal release of immunoreactive substance P. Following injection of kaolin and carrageenan into a knee, evidence for release of substance P following joint stimulation was found in 7 of 10 cats. Such release did not occur for several hours after joint injection and was detected predominantly in the superficial dorsal horn, the dorsal columns and at the dorsal surface of the spinal cord. In some experiments release was detected in the deep dorsal horn and upper ventral horn. Release of immunoreactive substance P required periods of mechanical stimulation such as flexion of, or pressure to, the inflamed joint. The failure to detect central release of substance P from stimulation of normal joints, and the release of substance P, after a delay, from inflamed joints, suggest that the fibres releasing this compound require sensitization by inflammatory mediators before they are excited by joint stimuli.

Neuropeptide Y: A putative neurotransmitter

Since its isolation in 1982, neuropeptide Y (NPY) has received considerable interest. This 36 amino acid peptide has been identified widely throughout the central and peripheral nervous systems, and within the autonomic system it appears in close association but not exclusively within catecholamine containing nerves. NPY begins to meet some of the criteria required to be established as a neurotransmitter. Thus, the peptide has been localised exclusively within nerves, and electron microscopy has shown NPY within nerve terminals. High affinity, saturable binding sites for NPY have been demonstrated in rat brain membranes, and the peptide has been reported to be released into the circulation during sympathetic nerve stimulation. The peptide is pharmacologically active both within the central nervous system by altering blood pressure, feeding and anterior pituitary function and in the periphery where NPY acts as a vasoconstrictor.

Differences in regional brain concentrations of neuropeptide Y in spontaneously hypertensive (SH) and Wistar-Kyoto (WKY) rats

Regional brain concentrations of neuropeptide Y immunoreactivity (NPY) were measured in age-matched Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rats using a sensitive and specific radioimmunoassay developed within our laboratory. In 5 of the 9 brain regions examined the SH rats had significantly lower NPY levels compared to the WKY strain. The largest differences occurred within the cortex (-43%), and cervical (-30%) and thoracic spinal cord (-30%), whilst smaller differences were observed in the midbrain (-11%) and medulla oblongata-pons (-18%). The concentrations of NPY in the hypothalamus and hippocampus did not vary between the strains. The SH rats contained significantly greater (+18%) NPY levels in the striatum compared to the WKY rats.

Nitric oxide increases interstitial excitatory amino acid release in the rat dorsomedial medulla oblongata

In vivo microdialysis was employed to measure release of the neuroactive amino acids L-glutamate (Glu) and L-aspartate (Asp) in the dorsomedial medulla oblongata of the anaesthetised rat. Basal levels of endogenous extracellular Glu and Asp were increased over 5-fold and 3-fold, respectively, following perfusion with a depolarising stimulus of KCl. Intracerebral administration of the nitric oxide (NO) donor S-nitroso-N-acetylpenicillamine (SNAP, 30 microM) caused a 5-fold increase in extracellular Asp and a 2-fold increase in extracellular Glu, which was blocked by Methylene blue. These data suggest that NO, acting through guanylate cyclase, can affect excitatory amino acid neurotransmission in the dorsomedial medulla oblongata.

Inflammatory Cell Infiltration after Endothelin-1-Induced Cerebral Ischemia: Histochemical and Myeloperoxidase Correlation with Temporal Changes in Brain Injury

Accumulation of neutrophils in brain after transient focal stroke remains controversial with some studies showing neutrophils to be deleterious, whereas others suggest neutrophils do not contribute to ischemic injury. Myeloperoxidase (MPO) has been used extensively as a marker for quantifying neutrophil accumulation, but is an indirect method and does not detect neutrophils alone. To elucidate the interaction of macrophages in the neutrophil inflammatory response, we conducted double-label immunofluorescence in brain sections at 0, 1, 2, 3, 7, and 15 days after ischemia. Each of these results was obtained from the same animal to determine correlations between neutrophil infiltration and ischemic damage. It was found that MPO activity increased up to 3 days after cerebral ischemia. Dual-staining revealed that macrophages engulf neutrophils in the brain and that this engulfment of neutrophils increased with time, with 50% of neutrophils in the brain engulfed at 3 days and approximately 85% at 15 days (N=5, P < 0.05). Interestingly, at 7 days the amount of dual-staining was decreased to 20% (N=5, P < 0.05). Neutrophil infiltration was positively correlated with ischemic damage in both the cortex and striatum (r2 = 0.86 and 0.80, respectively, P < 0.01). The results of this study indicate that the MPO from neutrophils phagocytized by macrophages may continue to contribute to the overall MPO activity, and that previous assessments that have utilized this marker to measure neutrophil accumulation may have miscalculated the number of neutrophils within the ischemic territory and hence their contribution to the evolution of the infarct at later time points. Thus any biphasic infiltration of neutrophils may have been masked by the accumulation of macrophages.

Medial prefrontal cortical lesions modulate baroreflex sensitivity in the rat

Previous neuroanatomical studies in rats have demonstrated that the medial prefrontal cortex sends projections to the nucleus of the solitary tract which also receives the bulk of baroreceptor information from primary afferents within the IXth and Xth cranial nerves. The present study examines the influence of the prefrontal cortex on baroreceptor heart rate reflex in conscious rats. Baroreceptor reflex activity was examined in rats with bilateral excitotoxin (N-methyl-D-aspartate)-induced lesions of the medial prefrontal cortex and in control rats (artificial cerebrospinal fluid). Seventeen to eighteen days after lesioning, reflex heart rate responses were recorded following intravenous bolus doses of the pressor agent phenylephrine and the depressor agent sodium nitroprusside. Baroreceptor reflex parameters i.e., maximum and average baroreceptor reflex gain (or sensitivity): minimum and maximum heart rate plateaus; heart rate range; upper and lower reflex thresholds, were determined by sigmoidal computerized curve-fitting. Lesioning the medial prefrontal cortex did not affect resting mean arterial pressure and heart rate. However, the lesion reduced maximum and average baroreceptor reflex gain and produced a small reduction in lower reflex threshold. The other parameters were unaffected by the lesion. These observations suggest that although the medial prefrontal cortex does not exert a tonic influence on brainstem vasomotor neurons, there may be a descending excitatory projection from this brain region to medullary neurones involved in the baroreceptor reflex arc.

L-glutamate as a neurotransmitter at baroreceptor afferents: Evidence from in vivo microdialysis

In vivo microdialysis was employed to measure release of endogenous L-glutamate and L-aspartate in the region of the dorsomedial medulla oblongata including the medial nucleus tractus solitarius of the anaesthetized rat. Basal extracellular levels of these amino acids were stable and increased over two-fold when the perfusate was changed to a high KCl (80 mM) artificial cerebrospinal fluid. This high K(+)-evoked release was calcium-dependent, while basal levels were insensitive to removal of calcium ions from the perfusate. An intravenous infusion of phenylephrine, which elevated blood pressure, caused a marked increase of both spontaneous and evoked release of glutamate. In contrast aspartate efflux was not significantly altered. The present data provide evidence that the excitatory amino acids, glutamate and aspartate, serve a neurotransmitter function in the nucleus tractus solitarius of the rat. Furthermore, the increase in glutamate release following baroreceptor activation with phenylephrine suggests that glutamate may be a neurotransmitter at baroreceptor afferent nerve terminals within the nucleus tractus solitarius. On the other hand, aspartate appears to be possibly an inter-neuronal transmitter in this brain region.

The detection of β-adrenoceptors on murine lymphocytes

Abstract The β-adrenoceptor antagonist ( 125 I)-iodohydroxybenzyl pindolol was used to detect and characterise β-adrenoceptors on the surface of mouse lymphocytes by a direct radioligand receptor assay using intact viable cells. The binding was stereoselective and appeared to be with a β 2 -type receptor as shown by competition with a number of adrenergic reagents. Scatchard analysis of the ligand binding demonstrated approximately 3000 binding sites per cell with an average dissociation constant ( K d ) of 0.92 nM. Little variation in binding characteristics was observed between lymphocytes from individual mice of either sex or in 24 different inbred mouse strains. In addition no significant differences were noted between T and B enriched lymphocyte populations, i.e. there was no functional polymorphism of the β-adrenoceptor on mouse lymphocytes. Thymocytes carried a lower affinity receptor which may be related to their functional immaturity. By using antisera specific for murine cell surface alloantigens, the β-adrenoceptor was shown not to be associated with the H-2K, H-2D, Ia, Ly−1, −2, −3, −4, or −5, Tla or Thy antigens. However, specific antisera for the Ly-7 antigen did partially inhibit IHYP binding. The results show that lymphocytes have a β 2 -adrenoceptor which closely resembles the receptor on other tissues.