Richard C. Rogers

Cell death in models of spinal cord injury

Current treatments for acute spinal cord injury are based on animal models of human spinal cord injury (SCI). These models have shown that the initial traumatic injury to cord tissue is followed by a long period of secondary injury that includes a number of cellular and biochemical cascades. These secondary injury processes are potential targets for therapies. Continued refinement of rat and mouse models of SCI, along with more detailed analyses of the biology of the lesion in these models, points to both necrotic and apoptotic mechanisms of cell death after SCI. In this chapter, we review recent evidence for long-term apoptotic death of oligodendrocytes in long tracts undergoing Wallerian degeneration following SCI. This process appears to be related closely to activation of microglial cells. It is has been thought that microglial cells might be the source of cytotoxic cytokines, such as tumor necrosis factor-alpha (TNF-alpha), that kill oligodendrocytes. However, more recent evidence in vivo suggests that TNF-alpha by itself may not induce necrosis or apoptosis in oligodendrocytes. We review data that suggests other possible pathways for apoptosis, such as the neurotrophin receptor p75 which is expressed in both neurons and oligodendrocytes after SCI in rats and mice. In addition, it appears that microglial activation and TNF-alpha may be important in acute SCI. Ninety minutes after a moderate contusion lesion, microglia are activated and surround dying neurons. In an atraumatic model of SCI, we have now shown that TNF-alpha appears to greatly potentiate cell death mediated by glutamate receptors. These studies emphasize that multiple mechanisms and interactions contribute to secondary injury after SCI. Continued study of both contusion models and other new approaches to studying these mechanisms will be needed to maximize strategies for acute and chronic therapies, and for neural repair.

Oxytocin, oxytocin antagonist, TRH, and hypothalamic paraventricular nucleus stimulation effects on gastric motility.

The roles of thyrotropin releasing hormone (TRH) and oxytocin as central regulators of gastric motility were investigated. Picomolar (4 picomoles) quantities of TRH injected into the dorsal motor nucleus of the vagus (DMN) elicited a significant increase in gastric motility while the same quantity of oxytocin elicited a reduction in phasic contractile activity and tone. The action of these peptides mimics the excitatory and inhibitory effects of stimulating the paraventricular nucleus of the hypothalamus (PVN); it is likely that this hypothalamic structure regulates gastric function through its peptidergic connections with medullary vagal structures. This hypothesis is supported by our observations that injections of an oxytocin antagonist into the DMN produced a disinhibition of gastric motility and an increase in the motility evoked by subsequent PVN stimulation. Vagotomy eliminated all subsequent central effects on motility of these peptides.

Brainstem pathways responsible for oesophageal control of gastric motility and tone in the rat

1 Previous anatomical studies indicate that the nucleus of the solitary tract, pars centralis (NSTc) contains the neurones which receive vagal afferent input from the oesophagus. The purpose of the present study was to characterize the NSTc circuits in the medulla that may be responsible for oesophageal control of gastric motility. 2 Moderate balloon distension of the oesophagus of the rat (14–18 mmHg) provoked a significant reduction in gastric motility and tone recorded with strain gauges. This receptive relaxation effect was eliminated by bilateral lesions centred on the NSTc. 3 NSTc cells activated by oesophageal distension were labelled extracellularly and juxtacellularly with neurobiotin. NSTc neurones send axonal projections throughout the entire rostral‐caudal extent of the dorsal motor nucleus of the vagus (DMN). These NSTc‐DMN connections were confirmed by retrograde transport of neurobiotin from DMN to NSTc. NSTc neurones were observed with dendrites arborizing within the ependymal lining of the fourth ventricles. Thus, NSTc neurones may be in position to monitor blood‐borne or ventricular agents and to alter the function of gastric‐vago‐vagal reflexes in response to these stimuli. 4 Neurophysiological recordings identified two subpopulations of DMN neurones which may be either activated or inhibited by oesophageal distension. Neurones excited by oesophageal distension were located mainly lateral and caudal in the DMN; neurones inhibited by oesophageal stimulation were located in medial and rostral DMN. 5 Our neurobiotin tracing results verified earlier studies showing that the NSTc projects to the intermediate reticular nucleus and the compact division of the nucleus ambiguus. Additionally, we found that the NSTc may be involved in reciprocal connections with the anterior, rostrolateral NST. 6 These results suggest that the gastric relaxation evoked by oesophageal distension is critically dependent on intact brainstem vago‐vagal circuits. The NSTc, the recipient of oesophageal afferent projections from the vagus nerve, sends axons to the entire DMN, the source of parasympathetic control of the stomach. DMN neurones respond differentially to oesophageal distension, reinforcing the view that oesophageal afferents may provoke gastric relaxation by activating a vagal inhibitory pathway while simultaneously inhibiting a vagal excitatory pathway.

Vagal control of digestion: Modulation by central neural and peripheral endocrine factors

Vago-vagal reflex control circuits in the dorsal vagal complex of the brainstem provide overall coordination over digestive functions of the stomach, small intestine and pancreas. The neural components forming these reflex circuits are under significant descending neural control. By adjusting the excitability of the different components of the reflex, alterations in digestion control can be produced by the central nervous system. Additionally, the dorsal vagal complex is situated within a circumventricular region without an effective blood-brain barrier. As a result, vago-vagal reflex circuitry is also exposed to humoral influences which profoundly alter digestive functions by acting directly on brainstem neurons. Behavioral and endocrine physiological observations suggest that this humoral afferent pathway may significantly alter the regulation of food intake.

Thyrotropin-releasing hormone: effects on identified neurons of the dorsal vagal complex

Previous reports have demonstrated that intraventricular administration of thyrotropin-releasing hormone (TRH) markedly elevates parasympathetic efferent activity. The following study determined if this response could be attributed to an effect of TRH on the neurons in the dorsal motor nucleus of the vagus (DMN) and/or the nucleus tractus solitarius (NTS), the nuclei that comprise the dorsal vagal complex (DVC). Individual DMN or NTS units were identified electrophysiologically by using stimulating electrodes placed on the cervical vagus. Alterations in firing rate of identified cells in response to pressure injection of TRH (10-40 fmol in 10-40 pl) or equal volumes of artificial cerebrospinal fluid (ACSF) were monitored. Of the DMN cells that were responsive to TRH, all were excited, whereas all responsive NTS cells were inhibited by this peptide. TRH was characterized as potent and had long-lasting effects on cells in DMN and NTS. The action of TRH on both nuclei in the dorsal vagal complex may explain the powerful effects of this peptide on vagally mediated functions.

EFFECTS OF PREGNANCY AND PROGESTERONE METABOLITES ON REGULATION OF SYMPATHETIC OUTFLOW

1. Pregnancy is characterized by a 40% increase in blood volume and cardiac output, a decrease in arterial blood pressure and thus a substantial decrease in total peripheral resistance. The aims of the experiments described in this manuscript were: (i) to determine if pregnancy resulted in alterations in baroreflex control of sympathetic outflow; and (ii) to evaluate possible mechanisms for pregnancy‐induced changes in control of sympathetic outflow.

Intramedullary connections of the gastric region in the solitary nucleus: a biocytin histochemical tracing study in the rat

The local circuit neurons in the solitary nucleus that form part of a gastro-gastric vago-vagal reflex were examined using a biocytin/avidin-peroxidase histochemical tracing method in the male Long-Evans rat. Iontophoretic deposits of very small amounts of biocytin were made into the ventral commissural nucleus of the solitary tract (vcNTS) where the excitatory neuronal response to antral distension was recorded. The tracing study revealed substantial axonal projections from the vcNTS to the immediately subjacent dorsal motor nucleus of the vagus (DMN) as well as the parvocellular reticular formation (pcRF). Some axons also appeared to terminate on neurons of the nucleus retroambiguus (nRAm) in the same coronal plane as the injection site. Labeled NTS neurons in the immediate area of the injection site revealed a clear horizontally-oriented pattern of dendrites, some of which extended from the midline to the solitary tract. Some of these dendrites could be found within the walls of arterioles, the central canal or in the area postrema. This finding suggests that vcNTS neurons activated by antral inflation are probably influenced by a number of other neural and chemical afferent signals.

Tumor Necrosis Factor-Alpha in the Dorsal Vagal Complex Suppresses Gastric Motility

Gastric hypomotility, loss of appetite, nausea, and vomiting frequently accompany critical infectious illness, radiation sickness, and carcinogenesis. The present studies examined the possibility that the pro-inflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), may be responsible for provoking some of these autonomic signs associated with illness. Gastric motility of urethane-anesthetized rats was prestimulated with intracisternal applications of thyrotropin-releasing hormone (TRH), a peptide known to activate parasympathetic vagal excitatory pathways to the stomach. Microinjection of TNF-alpha (as low as 0.02 fmol) directly into the dorsal vagal comples (DVC) suppressed TRH-stimulated gastric motility for prolonged periods of time. Duration of suppression ranged from 5 min to more than an hour, dependent on both the dose of TNF-alpha and accuracy of placement of the microinjection within the DVC. This suppression demonstrated a dose-dependent effect of TNF-alpha that required an intact vagal pathway. These studies indicate that TNF-alpha may represent a unique cytokine afferent signal which directly regulates the excitability of vago-vagal reflex circuits resulting in altered gastric motility during disease states.

Descending spinal projections from the rostral gigantocellular reticular nuclei complex

Electrophysiological and physiological studies have suggested that the ventral medullary gigantocellular reticular nuclei (composed of the gigantocellular ventralis and pars alpha nuclei as well as the adjacent lateral paragigantocellular nucleus; abbreviated Gi‐LPGi complex) provide descending control of pelvic floor organs (Mackel [1979] J. Physiol. (Lond.) 294:105–122; Hubscher and Johnson [1996] J. Neurophysiol. 76:2474–2482; Hubscher and Johnson [1999] J. Neurophysiol. 82:1381–1389; Johnson and Hubscher [1998] Neuroreport 9:341–345). Specifically, this complex of paramedian reticular nuclei has been implicated in the inhibition of sexual reflexes. In the present study, an anterograde fluorescent tracer was used to investigate direct descending projections from the Gi‐LPGi complex to retrogradely labeled pudendal motoneurons (MN) in the male rat. Our results demonstrated that, although a high density of arborizations from Gi‐LPGi fibers appears to be in close apposition to pudendal MNs, this relationship also applies to other MNs throughout the entire spinal cord. The Gi‐LPGi also projects to spinal autonomic regions, i.e., both the intermediolateral cell column and the sacral parasympathetic nucleus, as well as to regions of the intermediate gray, which contain interneurons involved in the organization of pelvic floor reflexes. Lastly, throughout the length of the spinal cord, numerous neurons located primarily in laminae VII–X, were retrogradely labeled with Fluoro‐Ruby after injections into the Gi‐LPGi. The diffuse descending projections and arborizations of this pathway throughout the spinal cord suggest that this brainstem area is involved in the direct, descending control of a variety of spinal activities. These results are in contrast with our observations of the discrete projections of the caudal nucleus raphe obscurus, which target the autonomic and somatic MNs involved specifically in sexual and eliminative functions (Hermann et al. [1998] J. Comp. Neurol. 397:458–474). J. Comp. Neurol. 455:210–221, 2003.

Nucleus raphe obscurus (nRO) influences vagal control of gastric motility in rats

Because the nucleus raphe obscurus (nRO) maintains a direct connection with the dorsal vagal complex in the medulla, this nucleus has the potential to influence vagal control of gastric function. Both electrical- and glutamate-induced activation of the nRO were found to enhance gastric motility and tone in the rat. The gastric responses to nRO stimulation were abolished by peripheral muscarinic blockade.