Biomedical subjects

T S Nowak

Publications and source records attributed to T S Nowak.

At least 37 records · Page 2Linked to original sources

PubMed1994

The heat shock/stress response in focal cerebral ischemia.

Focal ischemia results in striking changes in gene expression. Induction of hsp72, a member of the family of 70 kDa heat shock/stress proteins is a widely studied component of the generalized cellular response to injury known as the 'stress response' that is detected in brain after ischemia and other insults. This overview summarizes observations on hsp72 expression in models of focal cerebral ischemia, considering its cellular distribution, factors affecting its transcriptional and translational expression, and its potential relevance to post-ischemic pathophysiology. Hsp72 expression is essentially limited to regions in which cerebral blood flow falls below 50% of control levels, provided that residual perfusion allows synthesis of the induced mRNA and protein. The cellular distribution of hsp72 depends on the nature of the ischemic insult, with preferential vascular expression in severely ischemic territory that is destined to necrose, pronounced neuronal expression throughout the ischemic 'penumbra', and limited glial involvement in a narrow zone immediately surrounding the infarct. Together with results in other injury models, these observations indicate that hsp72 induction identifies discrete populations of surviving cells that are metabolically compromised, but not irreversibly damaged after focal ischemia. Available evidence suggests that the stress response is an important component of cellular defense mechanisms, and that successful accumulation of hsp72 is critical to survival following ischemia. Its expression may also contribute to mechanisms of induced ischemic tolerance. Future studies may be expected to more fully characterize the range of altered gene expression in response to focal ischemic injury and to establish specific roles for hsp72 and other induced proteins in the progression of injury and recovery following such insults.

Animals↗

PubMed1993

Kynurenine 3-hydroxylase in brain: species activity differences and effect of gerbil cerebral ischemia.

We have developed a rapid and highly sensitive assay for brain kynurenine 3-hydroxylase activity. The present study determined some characteristics and species differences of kynurenine 3-hydroxylase activity in brain and evaluated the response of this enzyme to cerebral ischemia. The kynurenine 3-hydroxylase assay is based on the conversion of L-kynurenine to 3-hydroxykynurenine in vitro and the quantification of 3-hydroxykynurenine by high-performance liquid chromatography. Kynurenine 3-hydroxylase activity was detected in human, macaque, rat, mouse, and gerbil brain. Regional gerbil brain activities ranged from 20 to 50 nmol/g/h, while kynurenine 3-hydroxylase activities in other species were one order of magnitude lower. Kynurenine 3-hydroxylase was also detected in lung, kidney, spleen, intestine, and liver of gerbils, with activities larger than in brain. Delayed increases in the activity of kynurenine 3-hydroxylase occur in several brain regions following transient ischemia in gerbils. These changes are particularly marked in regions that showed the most extensive brain damage (hippocampus and striatum). Kynurenine 3-hydroxylase may have an important role in determining the flux of kynurenine in brain.

Animals↗

PubMed1993

Synthesis of heat shock/stress proteins during cellular injury.

Several conclusions can be drawn from available data on the expression of stress proteins in brain with respect to their utility as markers of cellular injury. First, it is evident that all cell types in brain are capable of expressing stress proteins, although there is striking specificity in the population responding to a given insult. The apparent hierarchy of responsiveness indicated by hsp72 expression correlates well with the relative vulnerability of specific cell populations in a given model. With increasing severity of injury there can be an attenuation of the translational component of the stress response, in that hsp72 immunoreactivity fails to accumulate even though its mRNA is abundantly expressed. For this reason, hsp72 immunoreactivity provides an index of cell populations that have responded to an insult with a functional stress response. Such a response is not sufficient to guarantee survival, since many CA1 neurons that show significant hsp72 staining are eventually lost after global ischemia in the rat. However, brief insults that result in expression of hsp72 and other proteins encoded by induced mRNAs do result in tolerance to subsequent insults. Future studies may be expected to reveal the contributions of specific gene products to the tolerant state. Meanwhile, complementary evaluations of hsp72 mRNA and protein expression provide practical means of identifying cell populations responding to diverse injuries.

Animals↗

PubMed1993

Stress protein and proto-oncogene expression as indicators of neuronal pathophysiology after ischemia.

Induction of hsp70 mRNA and protein appear to provide useful markers for delineating stages in the progression of neuronal pathophysiology after ischemia. Detection of hsp70 encoded by the induced mRNA is dependent on complex interactions between the time course of mRNA expression and recovery of protein synthesis in a given neuron population, and perhaps other factors relating to specific aspects of hsp70 physiology, during recirculation intervals of hours to days. Transient mRNA expression and subsequent detection of immunoreactive hsp70 protein appear to identify neurons more likely to survive ischemia and other insults, while prolonged expression of hsp70 mRNA is associated with more severe neuronal injury. Fos and Jun immunoreactivities are also increased after ischemia, and provide indexes of functional gene expression during earlier recirculation periods. The accumulation of Fos immunoreactivity in particular designates neurons in which rapid recovery of protein synthesis during 1-3 h recirculation has allowed translation of the very transiently expressed c-fos mRNA. Jun-like immunoreactivity allows an evaluation of events at later recirculation intervals, and provides a clear demonstration of synthesis and accumulation of induced protein in CA1 neurons at 6 h following 2 min ischemia. Detailed understanding of the significance of such interactions between transcriptional and translational events will continue to evolve as information accumulates regarding the expression of additional mRNAs and proteins after ischemia. The present demonstration that Jun-like immunoreactivity accumulates in CA1 neurons after brief ischemia indicates that widespread changes in gene expression, expected as a consequence of such primary effects on transcription factor activity, are likely to contribute to the phenomenon of induced ischemic tolerance and to other persistent changes in the brain following diverse insults.

Animals↗

PubMed1993

Cardiac arrest-induced complete cerebral ischaemia in the rat: dynamics of postischaemic in vivo calcium uptake and protein synthesis.

Dynamics of pathological changes following cardiac arrest induced cerebral ischaemia were related to the findings of double tracer autoradiography of 45Ca and 3H leucine uptake as respective indicators of ischaemic injury and metabolic disturbance. Abnormal calcium accumulation, determined by 45Ca uptake, was related to injured but still living neurons and to reactive glial elements. 45Ca autoradiography confirmed a high sensitivity to neuronal injury of the nucleus reticularis thalami (NRT), hippocampal CA1 pyramidal layer, inferior colliculus, ventral thalamic nucleus (VTN), caudate nucleus, and parietal cortex. 3H leucine incorporation revealed that an initially widespread inhibition of protein synthesis was followed by its considerable recovery. Observations concerning hippocampal CA1 sector and VTN suggested that a significant degree of protein synthesis, maintained at the late stage after postischaemic recovery, was related to survival and regeneration of neurons and not to the presence of glial elements.

Animals↗

PubMed1993

Mechanism of delayed increases in kynurenine pathway metabolism in damaged brain regions following transient cerebral ischemia.

Delayed increases in the levels of an endogenous N-methyl-D-aspartate receptor agonist, quinolinic acid (QUIN), have been demonstrated following transient ischemia in the gerbil and were postulated to be secondary to induction of indoleamine-2,3-dioxygenase (IDO) and other enzymes of the L-tryptophan-kynurenine pathway. In the present study, proportional increases in IDO activity and QUIN concentrations were found 4 days after 10 min of cerebral ischemia, with both responses in hippocampus > striatum > cerebral cortex > thalamus. These increases paralleled the severity of local brain injury and inflammation. IDO activity and QUIN concentrations were unchanged in the cerebellum of postischemic gerbils, which is consistent with the preservation of blood flow and resultant absence of pathology in this region. Blood QUIN and L-kynurenine concentrations were not affected by ischemia. Brain tissue QUIN levels at 4 days postischemia exceeded blood concentrations, minimizing a role for breakdown of the blood-brain barrier. Marked increases in the activity of kynureninase, kynurenine 3-hydroxylase, and 3-hydroxyanthranilate-3,4-dioxygenase were also detected in hippocampus but not in cerebellum on day 4 of recirculation. In vivo synthesis of [13C6]QUIN was demonstrated, using mass spectrometry, in hippocampus but not in cerebellum of 4-day postischemic animals 1 h after intracisternal administration of L-[13C6]tryptophan. However, accumulation of QUIN was demonstrated in both cerebellum and hippocampus of control gerbils following an intracisternal injection of 3-hydroxyanthranilic acid, which verifies the availability of precursor to both regions when administered intracisternally. Notably, although IDO activity and QUIN concentrations were unchanged in the cerebellum of ischemic gerbils, both IDO activity and QUIN content were increased in cerebellum to approximately the same degree as in hippocampus, striatum, cerebral cortex, and thalamus 24 h after immune stimulation by systemic pokeweed mitogen administration, demonstrating that the cerebellum can increase IDO activity and QUIN content in response to immune activation. No changes in kynurenic acid concentrations in either hippocampus, cerebellum, or cerebrospinal fluid were observed in the postischemic gerbils compared with controls, in accordance with the unaffected activity of kynurenine aminotransferase activity. Collectively, these results support roles for IDO, kynureninase, kynurenine 3-hydroxylase, and 3-hydroxyanthranilate-3,4-dioxygenase in accelerating the conversion of L-tryptophan and other substrates to QUIN in damaged brain regions following transient cerebral ischemia. Immunocytochemical results demonstrated the presence of macrophage infiltrates in hippocampus and other brain regions that parallel the extent of these biochemical changes.(ABSTRACT TRUNCATED AT 400 WORDS)

3-Hydroxyanthranilic Acid↗

PubMed1993

Kynurenine pathway enzymes in brain: responses to ischemic brain injury versus systemic immune activation.

Accumulation of L-kynurenine and quinolinic acid (QUIN) in the brain occurs after either ischemic brain injury or after systemic administration of pokeweed mitogen. Although conversion of L-[13C6]tryptophan to [13C6]-QUIN has not been demonstrated in brain either from normal gerbils or from gerbils given pokeweed mitogen, direct conversion in brain tissue does occur 4 days after transient cerebral ischemia. Increased activities of enzymes distal to indoleamine-2,3-dioxygenase may determine whether L-kynurenine is converted to QUIN. One day after 10 min of cerebral ischemia, the activities of kynureninase and 3-hydroxy-3,4-dioxygenase were increased in the hippocampus, but local QUIN levels and the activities of the indoleamine-2,3-dioxygenase and kynurenine-3-hydroxylase were unchanged. By days 2 and 4 after ischemia, however, the activities of all these enzymes in the hippocampus as well as QUIN levels were significantly increased. Kynurenine aminotransferase activity in the hippocampus was unchanged on days 1 and 2 after ischemia but was decreased on day 4, at a time when local kynurenic acid levels were unchanged. A putative precursor of QUIN, [13C6]anthranilic acid, was not converted to [13C6]QUIN in the hippocampus of either normal or 4-day post-ischemic gerbils. Gerbil macrophages stimulated by endotoxin in vitro converted L-[13C6]tryptophan to [13C6]QUIN. Kinetic analysis of kynurenine-3-hydroxylase activity in the cerebral cortex of postischemic gerbils showed that Vmax increased, without changes in Km. Systemic administration of pokeweed mitogen increased indoleamine-2,3-dioxygenase and kynureninase activities in the brain without significant changes in kynurenine-3-hydroxylase or 3-hydroxyanthranilate-3,4-dioxygenase activities. Increases in kynurenine-3-hydroxylase activity, in conjunction with induction of indoleamine-2,3-dioxygenase, kynureninase, and 3-hydroxyanthranilate-3,4-dioxygenase in macrophage infiltrates at the site of brain injury, may explain the ability of postischemic hippocampus to convert L-[13C6]tryptophan to [13C6]QUIN.

3-Hydroxyanthranilate 3,4-Dioxygenase↗

PubMed1992

Evidence for increased calcium influx across the choroid plexus following brief ischemia of gerbil brain.

Following 5 min ischemia of gerbil brain, unidirectional transfer of calcium from plasma to cerebrospinal fluid was measured quantitatively with 45CaCl2 at 4 days postischemic recirculation. 45Ca influx across the choroid plexus increased significantly from 0.0101 +/- 0.001 min-1 measured in sham-operated animals (n = 15) to 0.0294 +/- 0.002 min-1 determined in ischemic animals (n = 21; P less than 0.05). Histological examination of choroid plexus was carried out in Cresyl violet-stained sections from gerbils subjected to 5 min ischemia followed by 4 days (n = 6) and 7 days (n = 7) postischemic recirculation. Increased calcium transfer to cerebrospinal fluid was associated with cell damage of choroid plexus observed at 4 days postischemia. Endothelial choroid plexus injury was still detectable at 7 days after the ischemic insult suggesting a long-lasting pathomorphological process. Postischemic alterations in choroid plexus functions apparently expose brain tissue to much higher calcium influx into cerebrospinal fluid which, in turn, may contribute to calcium-related cell damage of the central nervous system.

Animals↗

PubMed1992

Expression of heat shock protein, HSP72, in the guinea pig and rat cochlea after hyperthermia: immunochemical and in situ hybridization analysis.

The induction of the heat shock protein, HSP72, was studied in the cochlea of guinea pigs and rats subjected to a hyperthermic stress. Analyses were done by immunoblotting and immunocytochemistry at 6 and 12 h after heat shock, using a commercially available monoclonal antibody (Amersham), and by in situ hybridization 1 h after heat shock using an oligonucleotide probe. In guinea pig immunoblots of the cochlea, HSP72 was present in both unstressed and heat stressed animals and immunocytochemistry did not reveal any difference of staining between them. As opposed to guinea pig, HSP72 was not found in unstressed rat cochlea. Heat shock induced HSP72 expression in most inner ear tissues of the rat examined by immunoblotting. Immunocytochemistry and in situ hybridization localized HSP72 synthesis in ganglion neurons, Schwann cells, spiral limbus, spiral ligament and stria vascularis. The strongest immunoreactivity and highest density of silver grains were seen in the stria vascularis. All blood vessels were strongly immunoreactive and were outlined with silver grains. These results show that HSP72 synthesis can be induced by hyperthermia in rat cochlea and suggest that this protein could be a useful marker for assessment of the effects of specific stresses in this organ.

Animals↗

PubMed1992

Global cerebral ischemia associated with cardiac arrest in the rat: I. Dynamics of early neuronal changes.

Light microscopic neuronal changes were studied in rats subjected to 10 min of global ischemia produced by compression of the major cardiac vessels. Observations of cresyl violet-stained sections revealed early changes involving predominantly GABAergic neurons in various locations. In rats killed 15 min after recirculation, the changes were characterized by the appearance of a clear peripheral zone with condensation of the remaining neuronal cytoplasm. After 1 h, these zones appeared to be compartmentalized into individual pearl-like vacuoles, especially prominent in the nucleus reticularis thalami. After 3 h, the cytoplasmic vacuoles disappeared and the neuronal changes, particularly in the cerebral cortex, striatum, hippocampus, and pars reticulata of the substantia nigra, consisted mainly of hyperchromasia or loss of Nissl substance. After 2 days, the cerebral cortex and thalamus contained occasional neurons with conspicuously large nucleoli. After 7 days, the hippocampus revealed an approximately 50% loss of CA1 pyramidal neurons, associated with intense microglial reactivity in the stratum radiatum, whereas the neuronal destruction was more complete in the nucleus reticularis thalami. Our observations suggest a possibility that early changes in GABAergic neurons may provide a period of neuronal disinhibition and thus contribute to an excitatory ischemic damage in regions connected by GABAergic circuitry.

Animals↗

PubMed1991

Localization of 70 kDa stress protein mRNA induction in gerbil brain after ischemia.

Induction of mRNA encoding the 70 kDa stress/heat shock protein, hsp70, was evaluated in post-ischemic gerbil brain by in situ hybridization using an oligonucleotide probe selective for stress-inducible members of this gene family. Expression of hsp70 sequences was most pronounced in hippocampal CA1 neurons that fail to accumulate immunoreactive hsp70 protein, and that are selectively lost following ischemia. Hybridizable RNA continued to be expressed in CA1 through at least 48 h, essentially until the onset of cell death in this model. In contrast, dentate granule cells and CA2 neurons destined to survive the insult showed transient induction of hsp70 mRNA during the first 24 h of recirculation that disappeared prior to the detection of maximal hsp70 immunoreactivity in these cell populations. Pretreatment with a single injection of MK-801 (10 mg/kg) considerably attenuated the induction of hsp70 mRNA in hippocampus at 6 h of recirculation, an effect apparently mediated by persistent drug-induced hypothermia. The drug did not prevent the later, selective appearance of hsp70 hybridization in CA1 neurons at 24 h, nor did it protect against the subsequent loss of these cells. These results demonstrate a prolonged postischemic stress response at the transcriptional level in vulnerable hippocampal neurons, and suggest its utility as a marker for neuronal pathophysiology associated with mechanisms mediating delayed neuronal death.

Animals↗

PubMed1990

Role of neuroexcitation in development of blood-brain barrier and oedematous changes following cerebral ischaemia and traumatic brain injury.

Potential involvement of neuroexcitatory mechanisms was studied in: 1) repetitive forebrain ischaemia in gerbils, 2) global cerebral ischaemia in rats and 3) cryogenic injury to the cerebral cortex in rats and gerbils. Uptake of 45Ca was used as a marker of injury, whereas ultrastructural localization of calcium was assessed with an oxalate-pyroantimonate method. The blood-brain barrier was evaluated with immunostaining for serum albumin. Changes in extracellular glutamate were estimated by microdialysis and an enzymatic cycling assay. Changes in water content were assessed by specific gravity measurements. Repetitive ischaemia of 3 x 5 min carotid occlusions produced a cumulative effect with regard to development of oedema and neuronal injury. This was associated with several-fold increments in glutamate release after repeated insults, whereas there was no apparent correlation with energy metabolism disturbances. Other studies revealed in all models a development of secondary foci distant to the primary impact of ischaemia or cold lesions, which were characterized by calcium accumulation in swollen dendrites, chronic neuronal changes and intraneuronal uptake of serum proteins, all of these changes being potentially compatible with involvement of neuroexcitatory mechanisms.

Animals↗

PubMed1990

In vivo hyperthermia induces expression of HSP70 mRNA in brain regions controlling the neuroendocrine response to stress.

Regional localization of HSP70 expression in brain of rats exposed to increased ambient temperatures was examined using in situ hybridization. In addition to the cerebellar granule cell layer and choroid plexus, selective hybridization was observed in the hippocampal dentate gyrus, paraventricular and dorsomedial hypothalamic nuclei, median eminence, and medial habenula. Apparently, cells in brain regions coordinating the neuroendocrine response to stress show a preferential induction of cellular stress proteins in response to heat.

Animals↗

PubMed1990

Delayed increases in regional brain quinolinic acid follow transient ischemia in the gerbil.

Excessive activity or release of excitatory amino acids has been implicated in the neuronal injury that follows transient cerebral ischemia. To investigate the metabolism of the endogenous excitotoxin, quinolinic acid, and its potential for mediating cell loss following ischemia, the concentrations of quinolinic acid, L-tryptophan, 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid were quantified in gerbil brain regions at different times after 5 or 15 min of ischemia induced by bilateral carotid artery occlusion. Significant elevation of brain tryptophan levels, accompanied by increased 5-hydroxyindoleacetic acid concentrations, occurred during the first several hours of recirculation, but regional brain quinolinic acid concentrations were found either to decrease or remain unchanged during the first 24 h after the ischemic insult. However, significant increases in quinolinic acid concentrations occurred in striatum and hippocampus at 2 days of recirculation after 5 min of ischemia. After a further 4 and 7 days, strikingly large increases in quinolinic acid concentrations were observed in all regions examined, with the highest levels observed in the hippocampus and striatum, regions that also show the most severe ischemic injury. These delayed increases in brain quinolinic acid concentrations are suggested to reflect the presence of activated macrophages, reactive astrocytes, and/or microglia in vulnerable regions during and subsequent to ischemic injury. While the results do not support a role for increased quinolinic acid concentrations in early excitotoxic neuronal damage, the role of the delayed increases in brain quinolinic acid in the progression of postischemic injury and its relevance to postischemic brain function remain to be established.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1990

70-kilodalton heat shock protein induction in cerebellar astrocytes and cerebellar granule cells in vitro: comparison with immunocytochemical localization after hyperthermia in vivo.

Induction of the 70-kDa heat shock protein, hsp70, was evaluated in cultured cerebellar astrocytes and granule cell neurons subjected to a hyperthermic stress, using a monoclonal antibody and an oligonucleotide probe that selectively recognize stress-inducible species of hsp70-related proteins and RNAs, respectively. Immunoblots of cultures enriched in either granule cells or astrocytes, and immunocytochemical localization studies in cocultures of these cell types, demonstrated that hsp70 induction was restricted to the astrocyte population. Amino acid incorporation experiments showed little difference in the loss and recovery of overall protein synthesis activity in these two cell types following transient hyperthermic stress. RNA blot hybridizations confirmed the preferential glial induction of hsp70. In vivo immunocytochemical studies in brains of adult rats following hyperthermia were consistent with earlier observations that suggested a primarily glial and vascular localization of the heat shock response in most brain regions, although the intense immunoreactivity in the cerebellar granule cell layer suggests that there is induction of hsp70 in these neurons under in vivo conditions. These results suggest the potential value of such defined cell cultures in identifying mechanisms responsible for differences in the heat shock response of various cell types in vitro, and in revealing factors that may account for the apparent absence of the stress response in cultured cerebellar granule cell neurons.

Animals↗

PubMed1990

Heat shock RNA levels in brain and other tissues after hyperthermia and transient ischemia.

A number of studies have demonstrated increased synthesis of heat shock proteins in brain following hyperthermia or transient ischemia. In the present experiments we have characterized the time course of heat shock RNA induction in gerbil brain after ischemia, and in several mouse tissues after hyperthermia, using probes for RNAs of the 70-kilodalton heat shock protein (hsp70) family, as well as ubiquitin. A synthetic oligonucleotide selective for inducible hsp70 sequences proved to be the most sensitive indicator of the stress response whereas a related rat cDNA detected both induced RNAs and constitutively expressed sequences that were not strongly inducible in brain. Considerable polymorphism of ubiquitin sequences was evident in the outbred mouse and gerbil strains used in these studies when probed with a chicken ubiquitin cDNA. Brief hyperthermic exposure resulted in striking induction of hsp70 and several-fold increases in ubiquitin RNAs in mouse liver and kidney peaking 3 h after return to room temperature. The oligonucleotide selective for hsp70 showed equivalent induction in brain that was more rapid and transient than observed in liver, whereas minimal induction was seen with the ubiquitin and hsp70-related cDNA probes. Transient ischemia resulted in 5- to 10-fold increases in hsp70 sequences in gerbil brain which peaked at 6 h recirculation and remained above control levels at 24 h, whereas a modest 70% increase in ubiquitin sequences was noted at 6 h. These results demonstrate significant temporal and quantitative differences in heat shock RNA expression between brain and other tissues following hyperthermia in vivo, and indicate that hsp70 provides a more sensitive index of the stress response in brain than does ubiquitin after both hyperthermia and ischemia.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1990

70-kDa heat shock protein and c-fos gene expression after transient ischemia.

Induction of messenger RNA encoding the 70-kDa heat shock or stress protein, hsp70, and the product of the proto-oncogene c-fos was evaluated in gerbil hippocampus by in situ hybridization at various recirculation intervals after 5 minutes of ischemia. Striking increases in c-fos RNA were observed in dentate granule cells within 15 minutes of recirculation and remained evident through 1 hour, returning to undetectable control levels by 3 hours. Modest c-fos hybridization was seen in CA1 and CA3 neurons during the same time course. These results are consistent with the rapid and transient stimulation-induced c-fos expression observed in many experimental systems. Hsp70 expression showed a longer time course, being strongly induced in all major hippocampal neuron populations within 3 hours and persisting for approximately 12 hours in dentate granule cells and through 24 hours in CA3 pyramidal neurons. Notably, the most prolonged expression of hsp70 RNA was observed in vulnerable CA1 neurons that minimally accumulate the immunoreactive protein, with hybridization detected essentially until the death of this cell population at 3-4 days. These studies demonstrate an overlapping distribution of hsp70 and c-fos expression in gerbil hippocampus after ischemia, although there are differences in time course and in the relative induction observed in different neuron populations. The transient increase in c-fos hybridization in dentate granule cells is identical to that seen in various seizure paradigms and provides further support for activation of hippocampal circuitry after ischemia. The prolonged time course of hsp70 messenger RNA expression in vulnerable CA1 neurons may provide a molecular correlate of proposed excitotoxic mechanisms mediating delayed neuronal death.

Animals↗

PubMed1990

Putative neuroexcitation in cerebral ischemia and brain injury.

Involvement of neuroexcitatory mechanisms in cerebral ischemia and brain injury was explored in experimental models of repetitive forebrain ischemia by temporary occlusion of carotid arteries in gerbils and cryogenic injury to the cerebral cortex in rats and gerbils. Our observations in these models revealed a pattern of injury that involved some anatomic structures outside the areas of direct ischemic or traumatic insult. Such foci of injury revealed conspicuously abnormal uptake of 45Ca associated with slight or moderate neuronal alteration, whereas severely injured areas showed no 45Ca uptake. Electron microscopic observations revealed a characteristic presence of calcium in swollen dendrites, closely resembling pictures obtained in neuroexcitatory conditions such as epileptic seizures. Abnormal uptake of 45Ca was associated with apparent blood-brain barrier changes characterized by intracytoplasmic uptake of extravasated albumin into the neurons. Protein synthesis assayed by in vivo [3H]leucine incorporation was reduced in regions showing calcium accumulation. Our observations suggest that neuroexcitation may play an important role in development of secondary and chronic changes after ischemic or traumatic brain insults.

Animals↗