Abstract
The distribution of heat shock proteins (HSP) HSP60, HSP73, HSP72 and HSP25 in the isoosmotic cortex and the hyperosmotic medulla of the rat kidney was investigated using Western blot analysis and immunohistochemistry. HSP73 was homogeneously distributed throughout the whole kidney. The level of HSP60 was high in the renal cortex and low in the medulla. HSP25 and HSP72 were present in large amounts in the medulla. Only low levels of HSP25 and almost undetectable amounts of HSP72 were found in the cortex. HSP25 exists in one nonphosphorylated and several phosphorylated isoforms. Western blot analysis preceded by isoelectric focussing showed that HSP25 predominates in its nonphosphorylated form in the outer medulla but in its phosphorylated form in cortex and inner medulla. Although this intrarenal distribution pattern was not changed during prolonged anaesthesia (thiobutabarbital sodium), a shift from the nonphosphorylated to the phosphorylated isoforms of HSP25 occurred in the medulla. The characteristic intrarenal distribution of the constitutively expressed HSPs (HSP73, HSP60, HSP25) may reflect different states of metabolic activity in the isoosmotic (cortex) and hyperosmotic (medulla) zones of the kidney. The high content of inducible HSP72 in the medulla most likely is a consequence of the osmotic stress imposed upon the cells by the high urea and salt concentrations in the hyperosmotic medullary environment.
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Angelidis CE, Lazaridis I, Pagoulatos GN (1991) Constitutive expression of heat-shock protein 70 in mammalian cells confers thermoresistance. Eur J Biochem 199:35–39
Beck F-X, Sone M, Dörge A, Thurau K (1992) Effect of loop diuretics on organic osmolytes and cells electrolytes in the renal outer medulla. Kidney Int 42:843–850
Bitar KN, Kaminski MS, Hailat N, Cease KB, Strahler JR (1991) HSP27 is a mediator of sustained smooth muscle contraction in response to bombesin. Biochem Biophys Res Commun 181:1192–1200
Borkan SC, Schwartz JH (1993) Heat stress protein (HSP) associated cytoprotection during ischaemia in vitro (Abstract). J Am Soc Nephrol 4:732
Borkan SC, Emami A, Schwartz JH (1993) Heat stress proteinassociated cytoprotection of inner medullary collecting duct cells from rat kidney. Am J Physiol 265:F333-F341
Burel C, Mezger V, Pinto M, Rallu M, Trigon S, Morange M (1992) Mammalian heat shock protein families. Expression and functions. Experientia 48:629–634
Cheng MY, Hard FU, Martin J, Pollock RA, Kalousek F, Neupert W, Hallberg EM, Hallberg RL, Horwich AL (1989) Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature 337:620–625
Ciocca DR, Oesterreich S, Chamness GC, McGuire WL, Fuqua SA (1993) Biological and clinical implications of heat shock protein 27, 000 (Hsp27): a review. J Natl Cancer Inst 85:1558–1570
Cohen DM, Wasserman JC, Gullans SR (1991) Immediate early gene and HSP70 expression in hyperosmotic stress in MDCK cells. Am J Physiol 261:C594-C601
Delhaye M, Gulbis B, Galand P, Mairesse N (1992) Expression of 27-kD heat-shock protein isoforms in human neoplastic and nonneoplastic liver tissues. Hepatology 16:382–389
Deshaies RJ, Koch BD, Werner Washburne M, Craig EA, Schekman R (1988) A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature 332:800–805
Emami A, Schwartz JH, Borkan SC (1991) Transient ischemia or heat stress induces a cytoprotectant protein in rat kidney. Am J Physiol 260:F479-F485
Hard FU (1991) Heat shock proteins in protein folding and membrane translocation. Semin Immunol 3:5–16
Huot J, Houle F, Landry J (1993) Possible involvement of an oxyradical-sensitive HSP27 kinase in cellular resistance to oxidative stress. In: Landry J, Tanguay R, Huot J (eds.) Third Fisher Winter National Symposium on Cellular and Molecular Biology, Ville du Lac Delage, Quebec, March 18–21. [Abst.] P38
Imamoto N, Matsuoka Y, Kurihara T, Kohno K, Miyagi M, Sakiyama F, Okada Y, Tsunasawa S, Yoneda Y (1992) Antibodies against 70-kD heat shock cognate protein inhibit mediated nuclear import of karyophilic proteins. J Cell Biol 119:1047–1061
Iwaki T, Iwaki A, Liem RK, Goldman JE (1991) Expression of alpha B-crystallin in the developing rat kidney. Kidney Int 40:52–56
Johnston RN, Kucey BL (1988) Competitive inhibition of hsp70 gene expression causes thermosensitivity. Science 242:1551–1554
Kampinga HH, Muller E, Brunsting JF, Heine L, Konings AW, Issels RD (1993) Association of HSP72 with the nuclear (TX-100-insoluble) fraction upon heating tolerant and non-tolerant HeLa S3 cells. Int J Hyperthermia 9:89–98
Kaur P, Welch WJ, Saklatvala J (1989) Interleukin 1 and tumour necrosis factor increase phosphorylation of the small heat shock protein. Effects in fibroblasts, Hep G2 and U937 cells. FEBS Lett 258:269–273
Komatsuda A, Wakui H, Imai H, Nakamoto Y, Miura AB, Itch H, Tashima Y (1992) Renal localization of the constitutive 73-kDa heat-shock protein in normal and PAN rats. Kidney Int 41:1204–1212
Kriz W, Elger M, Lemley K, Sakai T (1990) Structure of the glomerular mesangium: a biomechanical interpretation. Kidney Int 38:S2-S9
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Landry J, Chretien P, Laszlo A, Lambert H (1991) Phosphorylation of HSP27 during development and decay of thermotolerance in Chinese hamster cells. J Cell Physiol 147:93–101
Lavoie JN, Gingras-Brenton G, Tanguay RM, Landry J (1993) Induction of chinese hamster HSP27 gene expression in mouse cells confers resistance shock. J Biol Chem 268:3420–3429
Lavoie JN, Hickey E, Weber LA, Landry J (1993) Modulation of actin microfilament dynamics and fluid phase pinocytosis by phosphorylation of heat shock protein 27. J Biol Chem 268:24210–24214
Matsubara O, Kasuga T, Marumo F, Itoh H, Tashima Y (1990) Localization of 90-kDa heat shock protein in the kidney. Kidney Int 38:830–834
McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Medina R, Spokes K, Cantley L, Epstein FH (1993) Hypertonicity increases the message for heat-shock protein 60 in rat kidney papilla (Abstract). J Am Soc Nephrol 4: 114P
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Pfaller W (1982) Structure function correlation on rat kidney. In: Hild W, van Limborgh J, Ortmann R, Pauly JE, Schieber TH (eds) Advances in anatomy embryology and cell biology. Springer, Berlin Heidelberg New York, pp 1–106
Rapoport TA (1990) Protein transport across the ER membrane. Trends Biochem Sci 15:355–358
Riabowol KT, Mizzen LA, Welch WJ (1988) Heat shock is lethal to fibroblasts microinjected with antibodies against hsp70. Science 242:433–436
Sawczuk IS, Hoke G, Olsson CA, Connor J, Buttyan R (1989) Gene expression in response to acute unilateral ureteral obstruction. Kidney Int 35:1315–1319
Sheik-Hamad D, Garcia-Perez A, Ferraris JD, Peters EM, Burg MB (1994) Induction of gene expression by heat shock versus osmotic stress. Am J Physiol 267:F28-F34
Skowyra D, Georgopoulos C, Zylicz M (1990) The E. coli dnaK gene product, the HSP70 homolog, can reactivate heat-inactivated RNA polymerase in an ATP hydrolysis-dependent manner. Cell 62:939–944
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Von Hippel PH, Schleich T (1969) The effects of natural salts on the structural and conformational stability of macromolecules in solutions. In: Timaheff SN, Fasman GD (eds) Structure and stability of biological macromolecules. Dekker, New York, pp 417–574
Wilkinson JM, Pollard I (1993) Immunohistochemical localization of the 25 kDa heat shock protein in unstressed rats: possible functional implications. Anat Rec 237:453–457
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Müller, E., Neuhofer, W., Ohno, A. et al. Heat shock proteins HSP25, HSP60, HSP72, HSP73 in isoosmotic cortex and hyperosmotic medulla of rat kidney. Pflugers Arch. 431, 608–617 (1996). https://doi.org/10.1007/BF02191910
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DOI: https://doi.org/10.1007/BF02191910