Biomedical subjects

P Gass

Publications and source records attributed to P Gass.

At least 73 records · Page 4Linked to original sources

PubMed1995

Correlation between seizure intensity and stress protein expression after limbic epilepsy in the rat brain.

Induction of heat shock/stress proteins is a key feature of a universal mechanism of cellular defence to injury known as the "stress response". The present study investigated whether heat shock protein expression correlates with the extent of neuronal injury inflicted by increasingly intense seizure activity. Limbic epilepsy was elicited by injecting intraperitoneally 8, 10 or 12 mg/kg kainic acid in adult Sprague-Dawley rats, resulting in graded degrees of seizure intensity and duration that closely correlated with the respective dose. Stress protein expression was investigated by immunocytochemistry and western blot analysis of microdissected brain areas with specific antibodies directed against representative members of three major classes of stress proteins, i.e. heat shock protein 72, heat shock protein 90 and heat shock protein 27, respectively. Heat shock protein 72 was absent in the brains of control animals, but markedly induced after limbic seizures in neurons of the limbic system, cortex, striatum and thalamus, with peak levels at 24 h. An increasing degree of seizure intensity caused a graded increase of heat shock protein 72 levels with a sequence reflecting the rank order of kainic acid susceptible hippocampal subpopulations. In contrast to heat shock protein 72, heat shock protein 90 was markedly expressed and equally abundant in all brain areas of untreated control animals and at any time point investigated following limbic seizures. Heat shock protein 27 was not detected in the brain of untreated animals nor following epilepsy. The present investigations demonstrate that the induction threshold of heat shock protein 72 in specific neuronal subpopulations clearly correlates with seizure intensity and duration. In addition, our experiments also define a narrow range of heat shock protein 72 expression with an upper limit beyond which heat shock protein 72 synthesis sharply declines. These findings reflect the risk of hippocampal neurons to undergo limbic seizure induced neuronal degeneration. It remains to be determined whether heat shock protein 72 expression is only a valuable marker for reversible neuronal injury or actually confers a neuroprotective effect.

Animals↗

PubMed1995

Hypoglycemia-elicited immediate early gene expression in neurons and glia of the hippocampus: novel patterns of FOS, JUN, and KROX expression following excitotoxic injury.

In the hippocampus there is a graded vulnerability of neuronal subpopulations to hypoglycemia-induced degeneration, most likely due to excitotoxic activation of glutamate receptors. The present study was conducted to investigate whether the induction of transcription factors of the immediate early gene (IEG) family after hypoglycemia reflects these different grades of neuronal vulnerability. We studied the expression profile of seven IEG-coded proteins in the rat hippocampus following severe insulin-induced hypoglycemia with 30 min of EEG isoelectricity and various survival periods for up to 42 h after glucose replenishment. Immunocytochemistry was performed on vibratome sections with specific polyclonal antisera directed against c-FOS, FOS B, c-JUN, JUN B, JUN D, KROX-24, and KROX-20. To unequivocally define the type of glial cells showing IEG induction, we investigated coexpression of c-FOS and glial marker proteins (glial fibrillary acid protein [GFAP], OX-42) by confocal laser scanning microscopy. Up to 3 h after glucose replenishment, differential temporospatial induction of IEG-coded transcription factors of the FOS, JUN and KROX families were observed in moderately injured neuronal subpopulations, including the majority of dentate granule cells and CA3 neurons. At later time points, however, a delayed and persistent c-JUN expression was found in severely, but reversibly, injured CA1 neurons and in neurons in the immediate vicinity of irreversibly damaged neurons in the crest of the dentate gyrus. Similar to the results with experimental models of central and peripheral axotomy, selective c-JUN induction in these neurons may represent an initial event in the regeneration process of sublethally injured neurons. In contrast to other models of excitotoxic injury such as ischemia and epilepsy, marked glial c-FOS expression was restricted to astrocytes, as assessed by confocal laser scanning microscopy.

Animals↗

PubMed1995

Selective c-JUN expression in CA1 neurons of the gerbil hippocampus during and after acquisition of an ischemia-tolerant state.

The selective delayed neuronal death of CA1 pyramidal cells after transient global ischemia in the gerbil brain can be prevented by preconditioning with a short sublethal period of ischemia 1-7 days prior to a subsequent, usually lethal ischemia of 5 min duration. Since changes of neuronal gene expression may play a crucial role in this tolerance induction, we investigated the postischemic expression profile of the fos, jun and Krox transcription factor families. We have previously reported that a single 5 min period of cerebral ischemia does not cause a de novo synthesis of immediate early gene (IEG) encoded proteins in CA1 neurons. In the present study, two experimental groups of Mongolian gerbils were investigated: one group was subjected to a single tolerance-inducing 2.5 min period of ischemia by bilateral occlusion of the common carotid artery. The second (combined ischemia) group was subjected to 2.5 min of ischemia, followed by 5 min of ischemia 4 days later. Post-ischemic expression of c-FOS, FOS B, c-JUN, JUN B, JUN D and KROX-24 was investigated by in situ hybridization and immunocytochemistry up to 48 h of recirculation. In contrast to a single 5 min period of ischemia, 2.5 min caused a postischemic expression of c-JUN protein, but no other IEGs, in CA1 neurons (peak at 6 h). Similarly, a selective but delayed c-JUN expression (peak at 18 h) was observed in animals subjected to combined ischemia. These results indicate that the induction of an endogenous neuroprotective state in CA1 neurons is associated with the activation of a genetic program which involves the expression of specific transcription factors.

Animals↗

PubMed1994

Constitutive expression of heat shock protein 90 (HSP90) in neurons of the rat brain.

Immunoblot analysis, immunocytochemistry and immuno-electron microscopy were employed to study the expression of HSP90 protein in the adult rat brain, using a specific polyclonal antiserum. Immunoblot analysis demonstrated equal levels of HSP90 in microdissected extracts from hippocampus, cortex, striatum and cerebellum. Immunocytochemistry and immuno-electron microscopy provided evidence that HSP90 is markedly expressed throughout all neuronal subpopulations of the CNS but not in non-neuronal cells except ependyma and choroid plexus. At the ultrastructural level, HSP90 immunoreactivity was predominantly found in perikarya but to a lesser extent also in dendrites and nuclei. The constitutive expression of HSP90 in widespread neuronal cell populations suggests a functional role in the physiological molecular program of CNS neurons.

Animals↗

PubMed1994

Point mutation of glycine receptor alpha 1 subunit in the spasmodic mouse affects agonist responses.

Homozygotic spasmodic (spd/spd) mice suffer from a motor disorder resembling poisoning by the glycine receptor antagonist strychnine. Here, a point mutation was identified in the glycine receptor alpha 1 subunit gene of the spasmodic mouse which predicts an alanine-to-serine exchange at position 52 of the mature polypeptide. Upon expression in Xenopus laevis oocytes, alpha 1A52S receptor channels displayed reduced responses to glycine, beta-alanine and taurine when compared to recombinant alpha 1 glycine receptors. As glycine receptor content in spinal cord and native molecular weight appeared unaltered, this suggests that the spasmodic phenotype results from an altered neurotransmitter sensitivity of the mutant alpha 1A52S subunit.

Alanine↗

PubMed1994

Induction of c-Jun and suppression of CREB transcription factor proteins in axotomized neurons of substantia nigra and covariation with tyrosine hydroxylase.

In adult rats, the expression of transcription factor proteins c-Jun and CREB and their colocalization with tyrosine hydroxylase (TH) were investigated in neurons of the substantia nigra compacta (SNC) axotomized by stereotaxic unilateral transection of the medial forebrain bundle (MFB). Axotomized SNC neurons were identified by injection of the retrograde tracer horseradish-peroxidase-coupled-gold (HRP-gold) into the ipsilateral striatum 5 days prior to MFB transection. Nuclear c-Jun immunoreactivity (IR) appeared 36 h after MFB transection in SNC neurons, was maximal after 5 days, and declined after 10 days. c-Jun-IR was visible in HRP-gold-labeled SNC neurons, demonstrating that c-Jun is in fact expressed in axotomized neurons. The constitutively expressed CREB (calcium/cAMP response element-binding protein, syn. CREB-1) was present in apparently all neuronal and glial cells in the brains of untreated rats including those SNC neurons that coexpressed TH. Three days following MFB transection, the nuclear CREB-IR disappeared in the axotomized SNC neurons labeled by TH-IR and was almost completely absent after 20 days in this neuronal population. The TH-IR rapidly declined 5 days after MFB transection, and 10 and 100 days post-axotomy the number of TH-labeled neurons was reduced by 52 and 80%, respectively. During this period, the majority of surviving TH positive neurons coexpressed c-Jun but were immunonegative for CREB. Between 3 and 60 days following MFB transection, the number of CREB-labeled glial cell nuclei increased in the ipsilateral substantia nigra by about 80%. Concomitantly, expression of GFAP, a marker protein for astrocytes, was also enhanced whereas nuclear c-Jun-, JunD-, and c-Fos-IR did not change in glial cells. These findings demonstrate that c-Jun can be expressed in axotomized neurons during the absence of CREB and suggest a role of c-Jun in the transcriptional control of the TH gene.

Animals↗

PubMed1994

Concurrent immediate early gene induction by epileptic seizures in heterotopic cortical grafts and neocortex.

Cortical primordia of rat fetuses (gestation day 14) were stereotactically grafted into the rostral striatum of adult recipient rats. After 8 weeks, the transplants had developed into a highly differentiated population of mature neuroectodermal cells. Host rats were then subjected to 15 min of bicuculline-induced epileptic seizures or served as controls. Seizure-elicited immediate early gene (IEG) expression was investigated after various postictal survival times (up to 24 h), using immunocytochemistry with specific antisera against seven IEG encoded proteins (c-FOS, FOS B, c-JUN, JUN B, JUN D, KROX-24, KROX-20). Constitutive IEG expression in intra striatum grafted neocortical neurons was identical to that in the corresponding host neocortex. In particular, abundant KROX-24 and lack of c-JUN expression implies the establishment of synaptic contacts within the graft or with the host circuitry. Postictal expression kinetics of individual IEG encoded proteins within the transplants were strikingly similar to those seen in the neocortex in situ. c-FOS and KROX-24 were most rapidly induced, followed by c-JUN and JUN B, and a more delayed induction of FOS B, JUN D and KROX-20. Apart from a slightly prolonged c-FOS expression in grafts, individual transcription factors remained elevated for different time periods and showed a concurrent decline in transplants and in neocortex in situ. In conclusion, IEG induction in grafts closely paralleled that in the host neocortex but differed from the adjacent striatum which exhibited no c-JUN induction at any time point investigated. These results indicate that following an appropriate differentiation period, heterotopically grafted embryonic cortical neurons respond to extracellular stimuli with changes of gene expression that closely resemble the normal host cortex. This suggests development of a similar molecular phenotype, including proper acquisition and intracellular processing of information.

Animals↗

PubMed1994

The transcription factor CREB is not phosphorylated at serine 133 in axotomized neurons: implications for the expression of AP-1 proteins.

The present study has investigated whether nerve fiber transection alters the phosphorylation of serine at position 133 (Ser133) of the transcription factor CREB (phosphoCREB). Activation of CREB by phosphorylation has a major function in the control of gene transcription. PhosphoCREB was visualized by antisera that specifically react with an epitope comprising the phosphorylated Ser133 of CREB as well as of CREM and ATF1 proteins. In untreated rats, nuclear immunoreactivity (IR) of phosphoCREB was consistently visible, e.g. in the cortex, thalamic and hypothalamic compartments and central termination areas of primary somatosensory afferents. Transection of peripheral (sciatic nerve), cranial (hypoglossal and facial nerve) and central (medial forebrain bundle and mammillo-thalamic tract) nerve fibers did not increase phosphoCREB-IR in the axotomized neurons between 5 min and 30 days post-axotomy. In contrast, phosphoCREB-IR appeared after 24 h in glial cells adjacent to the axotomized motoneurons and persisted up to 4 weeks. This increase in glial phosphoCREB-IR was paralleled by enhanced expression of the CREB protein itself. Between 20 min and 24 h following sciatic nerve transection, the number of phosphoCREB labeled nuclei also increased in neurons of the ipsilateral superficial dorsal horn of lumbar L3-L5 spinal cord segments. These data suggest that phosphorylation of Ser133 in CREB/CREM/ATF1 proteins is not involved in the transcriptional control of early-response genes such as c-jun in axotomized neurons following nerve transection. This is in contrast to the reported phosphorylation of CREB and its trans-acting effects on immediate-early genes such as c-fos after transynaptic neuronal excitation.

Afferent Pathways↗

PubMed1994

High induction threshold for transcription factor KROX-20 in the rat brain: partial co-expression with heat shock protein 70 following limbic seizures.

The transcription factor KROX-20, unlike many other immediate early genes, is not expressed in the rat hippocampus after bicuculline induced generalized seizures. Since limbic seizures are a more injurious stimulus, the KROX-20 expression profile was investigated in adult rats subjected to kainic acid induced limbic epilepsy at postictal intervals up to 48 h. Immunocytochemistry was performed using a specific polyclonal antiserum. In the hippocampus a sequential induction was observed with peak levels attained in dentate gyrus at 3 h, in CA1 at 8 h and in CA3 between 8 and 24 h, respectively. In contrast, no KROX-20 induction was found in hilus neurons. Prominent neuronal KROX-20 induction was also detected in other areas of the limbic system, in particular in amygdala and piriform cortex, as well as non-limbic regions such as neocortex and striatum. As is the case with KROX-20, heat shock protein (HSP) 70, a reliable marker for reversible neuronal injury, has a high induction threshold. Though not inducible in the hippocampus by generalized seizures, it is expressed after limbic epilepsy. Therefore, co-expression of KROX-20 and HSP70 was studied by a double labeling technique using a monoclonal antibody directed against the inducible form of HSP70. Neuronal subpopulations with perfect co-expression such as hippocampal CA1 neurons contrasted with others demonstrating partial co-induction (cortical neurons) or lack of co-expression (hilus cells), indicating that different stimuli trigger the activation of these two inducible genes.

Animals↗

PubMed1994

The spastic mouse: aberrant splicing of glycine receptor beta subunit mRNA caused by intronic insertion of L1 element.

Mice homozygous for the spastic mutation (spa) suffer from a complex motor disorder resulting from reduced CNS levels of the adult glycine receptor isoform GlyRA, which is composed of ligand-binding alpha 1 and structural beta polypeptides. The beta subunit-encoding gene (Glyrb) was mapped near the spa locus on mouse chromosome 3. In spa/spa mice, aberrant splicing of the beta subunit pre-mRNA strikingly diminishes the CNS contents of full-length transcripts, whereas truncated beta subunit mRNAs accumulate. This is a result of exon skipping, which causes translational frameshifts and premature stop codons. Intron 5 of the spa Glyrb gene contains an L1 transposable element that apparently is causal for the aberrant splicing of beta subunit transcripts.

Amino Acid Sequence↗

PubMed1994

Stimulus-transcription coupling in focal cerebral ischemia.

Glutamate-mediated spreading depression is currently thought to be a key event in the pathogenesis of potential neuronal degeneration in the ischemic 'penumbra'. Glutamate receptor stimulation causes induction of transcription factors that belong to the class of immediate early genes (IEGs), thought to be involved in coupling neuronal excitation to target gene expression. Focal cerebral ischemia elicits a homogeneous expression of several IEGs, prominently in cortex. In the ischemic core, discrepancies are observed between mRNA and protein levels, due to a severe, persistent protein synthesis deficit, preventing the translation of IEG encoded mRNAs. Outside the ischemic core, widespread IEG expression occurs in the entire ipsilateral cortex at mRNA as well as at protein level. This homogeneous expression of transcription factors can be pinpointed to at least two different pathogenetic mechanisms by means of appropriate pharmacological antagonists. Prolonged IEG induction in the 'penumbra', an area in which neurons are metabolically compromised but not yet energy-depleted, cannot be suppressed by the administration of N-methyl-D-aspartate (NMDA) receptor antagonists. In contrast, short-lasting IEG induction in undamaged neurons remote from the ischemic territory, though also caused by ischemia-elicited spreading depression, can be blocked by NMDA receptor antagonists. In both areas, IEG expression identifies neurons destined to survive but is likely to be mediated by different signal transduction pathways, at the receptor, second messenger and/or the DNA level.

Animals↗

PubMed1993

Regionally selective stimulation of mitogen activated protein (MAP) kinase tyrosine phosphorylation after generalized seizures in the rat brain.

Immunoblot analysis using a phosphotyrosine-specific antibody was performed to investigate tyrosine phosphorylation of the mitogen activated protein (MAP) kinase in the rat brain. Epileptic seizures induced by systemic injection of bicuculline caused a rapid and transient stimulation of MAP kinase tyrosine phosphorylation in hippocampus and somatosensory cortex. This increase in tyrosine phosphorylation was markedly attenuated by the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801. In contrast, in the cerebellum, tyrosine phosphorylation of MAP kinase remained undetectable after bicuculline-induced seizures. These results demonstrate that generalized seizures stimulate tyrosine phosphorylation of MAP kinase in a regionally selective manner.

Animals↗

PubMed1993

JUN, FOS, KROX, and CREB transcription factor proteins in the rat cortex: basal expression and induction by spreading depression and epileptic seizures.

The expression of the nuclear c-JUN, JUN B, JUN D, c-FOS, FOS B, KROX-24, and CREB transcription factors was investigated in the cortex of adult rats by immunocytochemistry. The expression patterns were studied in untreated rats and up to 24 hours following topical application of 1 M KCl to the cortical surface (KCl) or i.v. injection of bicuculline (BIC). Topical KCl induced cortical spreading depression and systemic injection of bicuculline evoked generalized tonic-clonic seizures. In untreated rats, JUN B, c-FOS, and FOS B were expressed in a small number of neurons in the piriform, perirhinal, entorhinal, and insular cortex and in layers II, III, and VI of all neocortical areas. In contrast, c-JUN, JUN D, and KROX-24 were expressed in all cortical layers but with different intensities of immunoreactivity (IR): c-JUN-IR was generally weak and predominantly present in layers II, III, and VI. JUN D-IR was equally strong in all layers. KROX-24 showed a prominent expression in layers II, IV, and VI. The CREB protein exhibited a slight preponderance in layer II and piriform cortex. Following KCl or BIC, a strong induction was seen for c-FOS, JUN B, and KROX-24, whereas c-JUN, JUN D, and FOS B showed only a moderate increase compared to basal levels. Changes of CREB-IR could not be detected. The localization of induced JUN, FOS, and KROX proteins reflected the pattern of labelling in untreated animals but demonstrated a higher intensity of labelling and an increased number of immunoreactive nuclei. The intensity and persistence of IR as well as the number of labelled cells following BIC exceeded those following KCl. Following BIC, increased levels of FOS B and JUN D were still present after 24 hours. Counterstaining with cresyl-violet and GFAP, a marker for astrocytes, revealed that JUN, FOS, and KROX proteins were expressed in neurons but not in glial cell populations. The present data demonstrate that CREB, JUN, FOS, and KROX transcription factors exhibit a layer-specific expression in the cerebral cortex with only slight area-specific differences both in untreated rats and following stimulation with KCl and BIC. The expression of transcription factor proteins indicate complex molecular genetic changes in cortical neurons due to pathophysiological events such as seizure activity and spreading depression.

Activating Transcription Factor 1↗

PubMed1993

Spatiotemporal induction of immediate early genes in the rat brain after limbic seizures: effects of NMDA receptor antagonist MK-801.

Fos, jun and krox belong to multigene families coding for transcription factors. These cellular immediate early genes (IEGs) are thought to be involved in coupling neuronal excitation to changes of target gene expression. Immunocytochemistry with specific antisera was used to assess regional levels of six IEG-encoded proteins (c-Fos, Fos B, Krox-24, c-Jun, Jun B, Jun D) in the rat forebrain after kainic acid-induced limbic seizures. The results demonstrate a complex spatial pattern of IEG induction and/or suppression in limbic and non-limbic structures. The sequence of induction within hippocampal subpopulations was identical for all IEGs investigated, following the order dentate gyrus, CA1 and CA3, and irrespective of different temporal profiles for individual transcription factors. Since Fos and Jun proteins act via homo- and heterodimer complexes at specific DNA sites, our data imply that the postictal combinatorial changes of these dimers allow a sequential and differential regulation of target gene expression in specific forebrain regions. Pretreatment with the non-competitive NMDA receptor antagonist MK-801 did not affect kainate-induced expression of IEGs in the limbic system, indicating that IEG induction in these regions is mediated by high-affinity kainate and AMPA receptors rather than NMDA receptors. In contrast, MK-801 abolished IEG induction in the somatosensory cortex and striatum, suggesting that IEG expression in non-limbic neurons occurs transsynaptically and is mediated by NMDA receptors.

Animals↗

PubMed1993

Expression of the c-erbB-2-encoded oncoprotein and progesterone receptor in human meningiomas.

The present study investigated the expression of c-erbB-2 in 59 meningiomas, including different histological subtypes and anaplastic variants, by immunocytochemistry and molecular biological techniques. Immunohistochemistry using the monoclonal antibody FWP-51 directed against c-erbB-2-encoded oncoprotein gp185 demonstrated variable degrees of immunoreactivity in all meningiomas. The intensity of immunostaining correlated with the degree of expression as assessed by Western analysis in 28 meningiomas using polyclonal antiserum 21N. There was no correlation between the degree of expression and histological variants. Immunoreactivity of all meningiomas was distinctly less intense, however, than that of the human breast cancer cell line SK-BR-3, and slightly lower than that of brain metastases of breast and ovarian carcinomas that served as positive controls for both methods. By Southern analysis all meningiomas showed a single copy of the c-erbB-2 gene. Non-neoplastic arachnoid cap cells also exhibited c-erbB-2 expression and the degree of immunoreactivity was comparable with the majority of meningiomas. These data argue against an overexpression of c-erbB-2 in meningiomas, but rather indicate a cell-type-specific constitutive expression of the c-erbB-2 gene product in meningiomas and their putative progenitor cells. Since a subgroup of meningiomas is known to express progesterone receptors (PR), gp185 immunoreactivity was compared to the hormone receptor status using monoclonal antibody KD68. Fifty-six percent meningiomas showed PR immunoreactivity, but there was no statistically significant correlation with the degree of gp185 expression.

Adult↗

PubMed1993

The KROX-20 transcription factor in the rat central and peripheral nervous systems: novel expression pattern of an immediate early gene-encoded protein.

The KROX-20 protein (also termed EGR-2) is encoded by an immediate early gene cloned by cross-hybridization to the Drosophila melanogaster Krüppel gene. It belongs to a class of transcription factors with zinc finger motifs and binding activity to a transcriptional regulatory DNA element termed the early growth response consensus sequence. In the present study the temporospatial expression of KROX-20 was investigated in the central and peripheral nervous systems of normal rats and after various stimuli known to induce immediate early genes, including epileptic seizures, axotomy, pharmacological treatment with glutamate and alpha-adrenergic receptor antagonists, and peripheral noxious stimulation. Immunocytochemistry was performed with a specific polyclonal antiserum generated against a fusion protein containing KROX-20 sequences. In the central nervous system, KROX-20 protein demonstrated distinct constitutive nuclear expression in specific neuronal subpopulations of the cortex, septum, amygdala, olfactory bulb and hypothalamus. In addition, distinct cytoplasmic immunoreactivity was present in spinal and medullary motoneurons, dorsal root ganglion neurons and a few neuronal cell populations of midbrain and forebrain. In the CNS, KROX-20 was only induced by bicuculline-induced epileptic seizures. Topographically, the postictal increase of KROX-20 levels was restricted to areas with constitutive expression, such as cerebral cortex, fornix and amygdala. Induction of KROX-20 peaked at 4-8 h after onset of seizure activity. No increase in immunoreactivity was observed in the hippocampus, the brain region most severely affected by bicuculline-induced seizures. Transection of central and peripheral nerve fibers did not result in KROX-20 induction in axotomized neurons. However, KROX-20 was induced in Schwann-like cells after transection of the sciatic nerve. In contrast to KROX-20, KROX-24, a related transcription factor of the zinc finger family, was markedly induced in hippocampal and spinal neurons following seizures and peripheral noxious stimulation, respectively, as well as in CNS neurons following axotomy. Our data indicate that KROX-20 represents an immediately early gene product with basal expression in selected neuronal populations of the nervous system and a restricted inducibility after intentional stimuli.

Animals↗

PubMed1993

Induction and suppression of immediate early genes in specific rat brain regions by the non-competitive N-methyl-D-aspartate receptor antagonist MK-801.

The expression pattern of six different immediate early gene-encoded proteins was examined in the rat forebrain after intraperitoneal administration of MK-801, a non-competitive N-methyl-D-aspartate receptor antagonist, at doses of 3 mg/kg and 0.3 mg/kg, respectively. Following MK-801 treatment, the presence of c-FOS, FOS B, KROX-24, c-JUN, JUN B, and JUN D were investigated by immunocytochemistry with specific antisera at different time intervals up to 48 h. Selective induction of all six immediate early genes was found in layer III neurons of the posterior cingulate and retrosplenial cortex. More complex effects were observed in the neocortex: MK-801 did not influence constitutive expression of different FOS and JUN proteins, but caused marked induction of c-FOS, FOS B, JUN B and JUN D, mainly in layer IV, but also in layers V and VI. In contrast, strong neocortical constitutive expression of KROX-24 was almost abolished by MK-801 administration, and replaced by an expression pattern similar to that of FOS and JUN proteins. Subcortical areas such as the hypothalamus and thalamus demonstrated an induction of a subset of immediate early genes (c-fos, fos B, Krox-24, jun B). Injection of MK-801 caused the same distributional pattern of immediate early gene expression irrespective of the dose given, but the extent of changes was stronger after 3 mg/kg, and altered levels of immunoreactivity persisted longer. In many experimental paradigms, immediate early genes are induced by N-methyl-D-aspartate receptor-mediated mechanisms. This induction can readily be blocked by N-methyl-D-aspartate receptor antagonists like MK-801. Our data, however, indicate that MK-801 itself causes immediate early gene expression in specific neuronal populations. In the present study MK-801-elicited expression of immediate early gene-encoded proteins seems to identify reversibly injured neurons, mainly in layer III of the posterior cingulate and retrosplenial cortex. These neurons have previously been shown to be the principal target of N-methyl-D-aspartate receptor antagonist toxicity. Since immediate early gene induction precedes heat-shock protein expression as well as pathomorphological changes, and is induced in additional cortical cell populations, it seems to be a more rapid and more sensitive indicator of non-lethal neuronal injury.

Animals↗

PubMed1993

Differential transcription and translation of immediate early genes in the gerbil hippocampus after transient global ischemia.

Excitotoxic activation of glutamate receptors is thought to be a key event for the molecular pathogenesis of postischemic delayed neuronal death of CA-1 neurons in the gerbil hippocampus. Glutamate receptor stimulation also causes induction of transcription factors that belong to the class of immediate early genes. We examined the expression of six different immediate early genes in the gerbil hippocampus after transient global ischemia. Comparative analysis of c-fos and Krox-24 expression was carried out in the same animals at the transcriptional and translational level by in situ hybridization and immunocytochemistry. Postischemic synthesis of four additional immediate early gene (IEG)-encoded proteins (FOS-B, c-JUN, JUN-B, and JUN-D) was investigated by immunocytochemistry at recirculation intervals between 1 and 48 h. After 5 min of ischemia, transcription of c-fos and Krox-24 mRNA was induced in all hippocampal subpopulations with peak expression at 1 h after recirculation. In vulnerable CA-1 neurons, increased transcription of c-fos and Krox-24 was not followed by translation into protein. Induction of immediate early gene-encoded proteins was restricted to neuronal populations less vulnerable to brief ischemia and identified neurons that are targets of glutamate receptor-mediated neurotoxicity but that are destined to survive. Our data indicate an asynchronous synthesis and persistence of individual IEG-encoded proteins in these neurons. The staggered induction implies that combinatorial changes of transcription factors allow a differential postischemic regulation of target gene expression both spatially and over time.

Animals↗