Effect of thermal preconditioning on Hsp70 expression in the medulla oblongata and on hemodynamics during passive hyperthermia.

A short-term episode of elevated core body temperature that induces Hsp70 expression (thermal preconditioning) may protect against heatstroke during subsequent hyperthermia. The protective effects of thermal preconditioning may involve several cellular and immunological mechanisms and improvements in baroreflex sensitivity. To substantiate the hypothesis that the protective effect of thermal preconditioning also occurs in conditions with intact thermoregulation, we examined the evolution of spontaneous cardiovagal baroreflex sensitivity and the protective effect of Hsp70 expression after thermal preconditioning in nonanesthetized Wistar-Kyoto rats with implanted telemetric transmitters. In the baroreflex centers of the medulla oblongata, thermal preconditioning induced Hsp70 in perineuronal and perivascular oligodendrocytes, microglia, and endothelial cells but not in neurons. The maximal Hsp70 expression was detected 4 h after preconditioning, but a significant number of Hsp70-positive cells was still present 72 h after preconditioning. Increased c-Fos expression in the neurons of baroreflex centers was detectable only 4 h after preconditioning. The mean values of cardiovagal baroreflex sensitivity did not show significant differences during the 72-hour follow-up period after thermal preconditioning. Similarly, cardiovascular variability measures of the autonomic nervous system activity were also not significantly affected by thermal preconditioning. During passive hyperthermia, thermal preconditioning had no statistically significant effect on thermoregulation and the onset of arterial pressure decline. Our data suggest that thermal preconditioning induces a glial type of Hsp70 expression in the baroreflex centers of the medulla oblongata. However, this response was not associated with cardiovagal baroreflex sensitization and protection against hemodynamic instability during passive hyperthermia.

NMDA preconditioning and neuroprotection in vivo: delayed onset of kainic acid-induced neurodegeneration and c-Fos attenuation in CA3a neurons.

Intraventricular (i.c.v.) kainic acid (KA) causes an acute excitotoxic lesion to the CA3 region of rodent hippocampus. Recent evidence implicated c-fos gene in regulating neuron survival and death following an excitotoxic insult. In this study we attempted to prevent KA-induced damage in CA3 neurons with NMDA preconditioning, which produced a marked expression of c-fos in the hippocampus. NMDA (0.6-6 microg, i.c.v.) was injected to anesthetized rats alone or 1 h before KA (0.15 microg, i.c.v.). Following KA injection, vibratome sections were processed for immunohistochemistry/electron microscopy. c-Fos and Nissl staining were used to estimate the extent of neuronal excitation and damage, respectively. Quantitative evaluation of c-Fos-labeled cells showed significantly less c-Fos in CA3a than in neighboring CA3b and CA2 from 1 to 4 h after KA alone. Attenuation of expressed c-Fos in CA3a was accompanied by damage of neurons with more apoptotic than necrotic signs. NMDA preconditioning elevated CA3a c-Fos expression and at 1 and 2 h exceeded markedly that after KA alone. However, at 4 h after KA, NMDA-preconditioned c-Fos induction in CA3a diminished to the same level as that seen after KA alone. The onset of neuronal degeneration was delayed in similar way. While NMDA-induced c-Fos expression in CA3a could be blocked by MK-801 completely, MK-801 and CNQX were both without significant effect on KA-induced c-Fos expression and neuronal damage. In conclusion, inhibition of c-Fos expression and onset of neuronal damage in CA3a following icv KA injection might be transiently delayed by i.c.v. NMDA preconditioning.

Cerebral neurons and glial cell types inducing heat shock protein Hsp70 following heat stress in the rat.

In this chapter, the distribution of Hsp70 in brain cell types following whole body hyperthermia is reviewed. The prevalence of Hsp70 expression in oligodendrocytes, microglia, and vascular cells in this type of stress contrasts with scarcity of Hsp70 induction in astrocytes and most neurons of the hyperthermic brain. However, a similarity between hyperthermic- and arsenite-induced brain patterns of Hsp70 expression supports the view that denaturation of specific proteins plays a major role in the selectivity of glial/vascular expression also during hyperthermia in vivo. The mechanism of neuronal Hsp70 non-responsiveness in heat stress despite their ability to use Hsc70 in a partial heat stress response remains to be elucidated.

Propentofylline rapidly normalizes mitochondrial respiration in a gerbil low flow unilateral forebrain ischemia.

The effect of propentofylline on mitochondrial respiratory activity was assessed in gerbils after 30 min of unilateral forebrain ischemia and 5 min of incomplete reperfusion. Cerebral blood flow (CBF) measured by hydrogen clearance was reduced to 12.6+/-2.1 ml/100 g per min in the ischemic hemisphere. Propentofylline at 10 mg/kg applied intraperitoneally raised CBF to 26.7+/-3.4 ml/100 g per min (P<0.05) in ischemic areas, however, CBF remained substantially reduced in comparison to the flow in the non-ischemic hemisphere (52.9+/-4.1 ml/100 g per min). Mitochondrial ADP-stimulated as well as uncoupled respiration was significantly reduced by approximately 60% after 30 min of ischemia. Since ADP-unstimulated respiration was not reduced, the respiratory control ratio declined markedly. Five minutes after application of propentofylline all indices of the mitochondrial respiratory capacity were normalized despite persisting reduction in CBF.

Identification of cerebral neurons and glial cell types inducing heat shock protein Hsp70 following heat stress in the rat.

Heat shock proteins were recently recognized as molecular chaperones that besides their chaperoning function were also involved in processes of cell death and survival. Many types of neural cells were reportedly capable of expressing heat shock protein Hsp70 following heat stress in vitro. However, identification of cell types inducing Hsp70 protein in the hyperthermic brain is not clear. In this study, cerebral Hsp70 distribution was evaluated in anesthetized adult rats (urethane, 1.5 g/kg, i.p.) subjected to short-term hyperthermia (41.5 degrees C for 30 min). Detection of Hsp70 was achieved by an ABC technique in vibratome or paraffin sections combined with specific markers of glial cell types. Hsp70 appeared by 90 min, mainly in glial and vascular cells, with enhanced immunostaining by 4 h following hyperthermia. Higher numbers of Hsp70-positive cells were detected in the white matter and diencephalic region than in the cerebral cortex, especially over the shorter interval. Hsp70 was localized in many oligodendrocytes, double-labeled with lectin GSII, and some vessels. Microglia showed apparently less Hsp70/OX-42 double-labeled cells than the previous two cell types. In contrast, only a few Hsp70-stained cells were positive for astrocyte marker GFAP. In addition to glial/vascular Hsp70 staining, neuronal Hsp70 induction was observed only in discrete regions including the paraventricular, supraoptic, suprachiasmatic and other hypothalamic nuclei, and in amygdala. Prevailing heat-stress expression of Hsp70 in oligodendrocytes and vascular cells might render them less susceptible to the consequences of other types of cell stress and could be exploited to increase selectively their survival in pathological situations.