Gerlinda E. Hermann

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.

Kinetics of glucocorticoid response to restraint stress and/or experimental influenza viral infection in two inbred strains of mice

The murine model of influenza viral infection was used to evaluate the effects of restraint stress on pathogenesis and survival in inbred strains of mice. We recently reported that restraint stress was associated with an enhanced probability of survival in one strain of inbred mouse, DBA/2, and not in another, C57BL/6. Those studies suggested that the protective mechanism(s) of stress on mortality in the DBA/2 mice might be attributable to elevated levels of circulating glucocorticoids. Therefore, daily levels of plasma glucocorticoids were measured during influenza viral infection in both these strains. The present studies demonstrated that influenza infection itself is perceived as a stressor in both C57BL/6 and DBA/2 mice as evidenced by elevated plasma glucocorticoid levels within 48 h of infection. However, augmentation of glucocorticoid levels was not seen in the DBA/2 mice that were also subjected to restraint stress during the course of infection. Thus, corticosterone levels alone did not account for the enhanced survival seen in this group of animals.

Stress-induced glucocorticoid response modulates mononuclear cell trafficking during an experimental influenza viral infection

The migration, distribution, and localization of lymphoid cells throughout the body is critical to the efficiency and development of the immune response. This study examined the role of endogenous glucocorticoids in mononuclear cell (MNC) trafficking during the development of an immune response to infection by influenza A/PR8 virus. Accumulation of MNC in the draining lymph nodes and at the site of virus replication (lungs) was studied in infected mice, and infected mice subjected to a stressor (physical restraint). The glucocorticoid antagonist, RU486, was used to block the activity of endogenous corticosterone during development of the immune response. PR8-infected mice demonstrated an elevation in circulating corticosterone regardless of whether they were treated with RU486 or a placebo. Thus, some afferent signal associated with the infection, and/or the immune response to infection, activated the hypothalamic-pituitary-adrenal axis (HPA) and was not subject to negative feedback regulation. The initial accumulation of MNC in the draining lymph nodes and lungs during infection, however, was independent of the glucocorticoid response. Our previous studies demonstrated that virally infected animals subjected to physical restraint had highly elevated plasma corticosterone levels, suppressed lymphadenopathy, and reduced accumulation of MNC in the lungs. In the present study, RU486 treatment restored cellularity to the draining lymph nodes and enhanced accumulation of MNC in lungs of stressed, A/PR8 virus-infected mice.

Restraint stress differentially affects the pathogenesis of an experimental influenza viral infection in three inbred strains of mice

Genetic variation in the response to stress may play a critical role in susceptibility to inflammatory diseases and development of the immune response. Experimental influenza viral infection was used to study the effects of restraint stress (RST) on pathogenesis and development of the immune response. Three inbred strains of mice (C57BL/6, DBA/2, and C3H/HeN) were infected with influenza A/PR8 and subjected to repetitive cycles of RST during development of the immune response. RST diminished cellular immune and inflammatory responses in all three strains; yet only the DBA/2 strain demonstrated RST-associated reduction in influenza viral-induced mortality.

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.

STRESS-INDUCED CHANGES ATTRIBUTABLE TO THE SYMPATHETIC NERVOUS SYSTEM DURING EXPERIMENTAL INFLUENZA VIRAL INFECTION IN DBA/2 INBRED MOUSE STRAIN

The murine model of influenza viral infection was used to evaluate the effects of restraint stress on pathogenesis and survival in the DBA/2 inbred strain of mice. Restraint stress has been associated with an enhanced probability of survival during influenza infection in this strain of mouse. Previous studies suggested that the protective mechanism(s) of stress on mortality might be attributable to elevated levels of circulating glucocorticoids. Subsequent work demonstrated that corticosterone levels alone could not account for the enhanced survival seen in the DBA/2 mice. The present studies examined the role of catecholamines in behavioral stress during influenza infection. It appears that glucocorticoids may play a primary role in trafficking and restriction of inflammation, while catecholamines may play role in limiting activation of virus-specific effector cells. The studies presented here suggest that the interplay between these two physiological response mechanisms needs to be coordinated to optimize development of the immune response to an infection.