[en] In this study we investigated the time course, cell-type and stress-specific expression of hsp70 mRNA and Hsp70 protein in glial cells and neurons in the rat brain following heat shock treatment and kainic acid-induced status epilepticus. Transcripts for hsp70 were detected in hippocampal homogenates from 1.5 to 6 h following hyperthermia and from 3 to 24 h following kainic acid-induced seizures. In situ hybridization revealed hsp70 mRNA to be region specific and time-dependent following hyperthermia and kainic acid-induced seizures. Western analysis indicated that Hsp70 reached maximal levels at 3 h after hyperthermia and 12 h after kainic acid-induced seizures. Immunohistochemistry revealed low level expression of Hsp70 protein in dentate granule cells at 1.5 and 3 h after hyperthermia. No Hsp70 protein was detected in neurons of the pyramidal cell layer or dentate hilus at any time following hyperthermia. Small Hsp70-immunoreactive cells were detected throughout the hippocampus following hyperthermia that, based on cell size, distribution, and double-labeling with vimentin, were considered to be glia. In contrast, high levels of Hsp70 protein were detected in neurons of the pyramidal cell layer and dentate hilus at 24 h after seizure-inducing kainic acid injection. These results suggest that expression of Hsp70 protein is cell-specific depending on the stressor. In addition, finding high levels of Hsp70 mRNA in the dentate granule cells after hyperthermia, but little or no Hsp70 protein, suggests that the synthesis of the protein is also regulated at the post-transcriptional level following hyperthermia.
Armstrong J.N., Plumier J.-C.L., Robertson H.A., Currie R.W. The inducible 70,000 mol. wt heat shock protein is expressed in the degenerating dentate hilus and piriform cortex after systemic administration of kainic acid in the rat. Neuroscience. 74:1996;685-693.
Baldwin S.A., Scheff S.W. Intermediate filament change in astrocytes following mild cortical contusion. Glia. 16:1996;266-275.
Ben-Ari Y. Limbic seizure and brain damage produced by kainic acid: mechanisms and relevance to human temporal lobe epilepsy. Neuroscience. 14:1985;375-403.
Bignami A., Dahl D. Vimentin-GFAP transition in primary dissociated cultures of rat embryo spinal cord. Int. J. Dev. Neurosci. 7:1989;343-357.
Blake M.J., Gershon D., Fargnoli J., Holbrook N.J. Discordant expression of heat shock protein mRNAs in tissues of heat stressed rats. J. Biol. Chem. 265:1990;15257-15279.
Brown I.R., Rush S.J. Expression of heat shock genes (hsp70) in the mammalian brain: distinguishing constitutively expressed and hyperthermia-inducible mRNA species. J. Neurosci. Res. 25:1990;14-19.
Brown I.R. Induction of heat shock (stress) genes in the mammalian brain by hyperthermia and other traumatic events: a current perspective. J. Neurosci. Res. 27:1990;247-255.
Cammer W., Tansey F.A., Brosnan C.F. Gliosis in the spinal cord of rats with experimental allergic encephalomyelitis: immunostaining of carbonic anhydrase and vimentin in reactive astrocytes. Glia. 2:1989;223-230.
Currie R.W. Synthesis of stress-induced protein in isolated and perfused rat hearts. Biochem. Cell Biol. 64:1986;418-426.
Currie R.W., White F.P. Characterization of the synthesis and accumulation of a 71-kilodalton protein induced in rat tissues following hyperthermia. Can. J. Biochem. Cell Biol. 61:1983;438-446.
Currie R.W., White F.P. Trauma-induced protein in rat tissues: a physiological role for a "heat shock" protein? Science. 214:1981;72-73.
Dahl D., Rueger D.C., Bignami A. Vimentin, the 57 000 molecular weight protein of fibroblast filaments, is the major cytoskeletal component in immature glia. Eur. J. Cell Biol. 24:1981;191-196.
David J.C., Currie R.W., Robertson H.A. Expression and distribution of hsp71 and hsc73 messenger RNAs in rat brain following heat shock: effect of dizocilpine maleate. Neuroscience. 62:1994;945-954.
Dragunow M., Currie R.W., Robertson H.A., Faull R.L. Heat shock induces c-fos protein-like immunoreactivity in glial cells in adult rat brain. Exp. Neurol. 106:1989;105-109.
B.E. Dwyer, R.N. Nishimura, Heat shock and neuroprotection in the CNS culture, in: J. Mayer, I.R. Brown (Eds.), Heat Shock Protein in the Nervous System, Academic Press, London, 1994, pp. 101-121.
Foster J.A., Brown I.R. Differential induction of heat shock mRNA in oligodendrocytes, microglia, and astrocytes following hyperthermia. Mol. Brain Res. 45:1997;207-218.
J. Gehrmann, G.W. Kreutzberg, Microglia in experimental neuropathology, in: H. Ketterman, B.R. Ransom (Eds.), Neuroglia, Oxford Univ. Press, Oxford, 1995, pp. 883-904.
Gonzalez M.F., Shiraishi K., Hisanaga K., Sagar S.M., Mandabach M., Sharp F.R. Heat shock proteins as markers of neural injury. Mol. Brain Res. 6:1989;93-100.
Goto S., Korematsu K., Oyama T., Yamada K., Hamada J., Inoue N., Nagahiro S., Ushio Y. Neuronal induction of 72-kDa heat shock protein following methamphetamine-induced hyperthermia in the mouse hippocampus. Brain Res. 626:1993;351-356.
Harrub J.B., Nowak T.S. Jr. Cryptic expression of the 70-kDa heat shock protein, hsp72, in gerbil hippocampus after transient ischemia. Neurochem. Res. 23:1998;703-708.
Hopkins D.A., Plumier J.-C.L., Currie R.W. Induction of the 27-kDa heat shock protein (Hsp27) in the rat medulla oblongata after vagus nerve injury. Exp. Neurol. 153:1998;173-183.
Johnston R., Kucey B.L. Competitive inhibition of hsp70 gene expression causes thermosensitivity. Science. 242:1988;1551-1553.
Kinouchi H., Sharp F.R., Hill M.P., Koistinaho J., Sagar S.M., Chan P.H. Induction of 70-kDa heat shock protein and hsp70 mRNA following transient focal cerebral ischemia in the rat. J. Cereb. Blood Flow Metab. 13:1993;105-115.
Kinouchi H., Sharp F.R., Koistinaho J., Hicks K., Kamii H., Chan P.H. Induction of heat shock hsp70 mRNA and HSP70 kDa protein in neurons in the 'penumbra' following focal cerebral ischemia in the rat. Brain Res. 619:1993;334-338.
Laemmli U.K. Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature. 227:1970;680-685.
Li Y., Chopp M., Yoshida Y., Levine S.R. Distribution of 72-kDa heat shock protein in the rat brain after hyperthermia. Acta Neuropathol. 84:1992;94-99.
Longo F.M., Wang S., Narasimhan P., Zhang J.S., Chen J., Massa S.M., Sharp F.R. cDNA cloning and expression of stress-inducible rat hsp70 in normal and injured rat brain. J. Neurosci. Res. 36:1993;325-335.
Lowenstein D.H., Chan P.H., Miles M.F. The stress protein response in cultured neurons: characterization and evidence for a protective role in excitotoxicity. Neuron. 7:1991;1053-1060.
Lowry O.H., Rosebrough N.J., Farr A.L., Randall R.J. Protein measurement with folin phenol reagent. J. Biol. Chem. 193:1951;265-275.
Marcuccilli C.J., Mathur S.K., Morimoto R.I., Miller R.J. Regulatory differences in the stress response of hippocampal neurons and glial cells after heat shock. J. Neurosci. 16:1996;478-485.
Marini A.M., Kozuka M., Lipsky R.H., Nowak T.S. Jr. 70-Kilodalton heat shock protein induction in cerebellar astrocytes and cerebellar granule cells in vitro: comparison with immunocytochemical localization after hyperthermia in vivo. J. Neurochem. 54:1990;1509-1516.
McCabe T., Simon R.P. Hyperthermia induces 72 kDa heat shock protein expression in rat brain in non-neuronal cells. Neurosci. Lett. 159:1993;163-165.
Miller E.K., Raese J.D., Morrison-Bogorad M. Expression of heat shock protein 70 and heat shock cognate 70 messenger RNAs in rat cortex and cerebellum after heat shock or amphetamine treatment. J. Neurochem. 56:1991;2060-2071.
Morimoto R.I. Cells in stress: transcriptional activation of heat shock genes. Science. 259:1993;1409-1410.
R.I. Morimoto, D.A. Jurivich, P.E. Kroeger, S.K. Mather, S.P. Murphy, A. Nakai, K. Sarge, K. Abravaya, L.T. Sistonen, Regulation of heat shock gene transcription by a family of heat shock factors, in: R.I. Morimoto, A. Tissières, C. Georgopolous (Eds.), The Biology of Heat Shock Proteins and Molecular Chaperones, Cold Spring Harbour Laboratory Press, New York, 1994, pp. 417-455.
R.I. Morimoto, P.E. Kroeger, J.J. Cotto, The transcriptional regulation of heat shock genes: a plethora of heat shock factors and regulatory conditions, in: U. Feige, R.I. Morimoto, I. Yahara, B. Polla (Eds.), Stress-Inducible Cellular Responses, Birkhaüser Verlag, Basel, Switzerland, 1996, pp. 139-163.
Morimoto R.I., Sarge K.D., Abravaya K. Transcriptional regulation of heat shock genes: a paradigm for inducible genomic responses. J. Biol. Chem. 267:1992;21987-21990.
Morrison-Bogorad M., Pardue S., McIntire D.D., Miller E.K. Cell size and the heat-shock response in rat brain. J. Neurochem. 63:1994;857-867.
Nishi S., Taki W., Uemura Y., Higashi T., Kikuchi H., Kudoh H., Satoh M., Nagata K. Ischemic tolerance due to the induction of HSP70 in a rat ischemic recirculation model. Brain Res. 615:1993;281-288.
Nishimura R.N., Dwyer B.E., Clegg K., Cole R., de Vellis J. Comparison of the heat shock response in cultured cortical neurons and astrocytes. Mol. Brain Res. 9:1991;39-45.
Nishimura R.N., Dwyer B.E., Vinters H.V., de Vellis J., Cole R. Heat shock in cultured neurons and astrocytes: correlation of ultrastructure and heat shock protein synthesis. Neuropathol. Appl. Neurobiol. 17:1991;139-147.
Nowak T.S. Jr., Bond U., Schlesinger M.J. Heat shock RNA levels in brain and other tissues after hyperthermia and transient ischemia. J. Neurochem. 54:1990;451-458.
Nowak T.S. Jr., Ikeda J., Nakajima T. 70-kDa heat shock protein and c-fos gene expression after transient ischemia. Stroke. 21:1990;III107-III111. 11 Suppl.
Olney J.W., Rhee V., Ho O.L. Kainic acid: a powerful neurotoxic analogue of glutamate. Brain Res. 77:1974;507-512.
Planas A.M., Soriano M.A., Estrada A., Sanz O., Martin F., Ferrer I. The heat shock stress response after brain lesions: induction of 72 kDa heat shock protein (cell types involved, axonal transport, transcriptional regulation) and protein synthesis inhibition. Prog. Neurobiol. 51:1997;607-636.
Plumier J.-C.L., Armstrong J.N., Landry J., Babity J.M., Robertson H.A., Currie R.W. Expression of the 27,000 mol wt heat shock protein following kainic acid-induced status epilepticus in the rat. Neuroscience. 75:1996;849-856.
Plumier J.-C.L., David J.-C., Robertson H.A., Currie R.W. Cortical application of potassium chloride induces the low molecular weight heat shock protein (Hsp27) in astrocytes. J. Cereb. Blood Flow Metab. 17:1997;781-790.
Plumier J.-C.L., Krueger A.M., Currie R.W., Kontoyiannis D., Kollias G., Pagoulatos G.N. Transgenic mice expressing the human inducible Hsp70 have hippocampal neurons resistant to ischemic injury. Cell Stress Chaperon. 2:1997;162-167.
Schroeter M., Schiene K., Kraemer M., Hagemann G., Weigel H., Eysel U.T., Witte O.W., Stoll G. Astroglial responses in photochemically induced focal ischemia of the rat cortex. Exp. Brain Res. 106:1995;1-6.
Sloviter R.S., Lowenstein D.H. Heat shock protein expression in vulnerable cells of the rat hippocampus as an indicator of excitation-induced neuronal stress. J. Neurosci. 12:1992;3004-3009.
Sloviter R.S., Nilaver G. Immunocytochemical localization of GABA-, cholecystokinin-, vasoactive intestinal polypeptide-, and somatostatin-like immunoreactivity in the area denta and hippocampus of the rat. J. Comp. Neurol. 256:1987;42-60.
W.J. Streit, Microglia cells, in: H. Kettenman, B.R. Ransom (Eds.), Neuroglia, Oxford Univ. Press, Oxford, 1995, pp. 85-97.
Vass K., Berger M.L., Nowark T.S. Jr., Welch W.J., Lassmann H. Induction of stress protein hsp70 in nerve cells after status epilepticus in the rat. Neurosci. Lett. 100:1989;259-264.