Different Responses in Brain Regions upon Heat Shock in Adult Zebrafish (Danio rerio)

  • Hwang, Chang-Nam (Division of Life Sciences, School of Life Sciences and Biotechnology, Korea University) ;
  • Lee, Dong-Ho (Division of Life Sciences, School of Life Sciences and Biotechnology, Korea University) ;
  • Lee, Sang-Ho (Division of Life Sciences, School of Life Sciences and Biotechnology, Korea University)
  • Published : 2009.09.30

Abstract

HSP70 has widely been induced in in vivo hyperthermia conditions in various organisms to study gene regulation and recently neuroprotectve roles of the induced gene expression under varying conditions. We investigated different responses among various tissues in zebrafish under heat shock to evaluate whether spatial and temporal expression pattern of zebrafish (z) hsp70 in transcriptional and translational level under heat shock stress in different brain regions. Heat shock groups were given for 1 h at $37^{\circ}C$ after recovery by transferring the treated animals back to $28^{\circ}C$ for 1, 2 and 24 h for recovery, respectively. Control (CTRL) group was kept at $28^{\circ}C$. At the end of treatments, five animals were collected and used for isolation of total RNAs and peptides from the corresponding tissues. Expression of zhsp70 mRNA showed different patterns in recovery periods in the tissues including the brain, eye, intestines, muscles, heart and testis by RT-PCR. Unlike the RT-PCR analysis, Northern blot analysis demonstrated nearly 30-fold increase in zhsp70 at 1 h heat shock, suggesting that RT-PCR may not be appropriate in unmasking regulation of the time-dependent zhsp70 expression. In the experiment involving different brain regions, the cerebellum showed gradual activation at 1 h to R1h and decreases in R2h and R24h, while the medulla oblongata and optic tectum showed gradual increase at R1h and decrease at R24h, indicating that different brain tissues respond specifically to heat shock in inducing zhsp70 and recovering from the heat shock status. Western blot analysis also demonstrated that the intracellular levels of zHSP70 in three different brain regions including the cerebellum, medulla oblongata and optic tectum are differently induced and recovered to normal state. These results clearly demonstrate that different regions of the body and the brain tissues are responding differently to heat shock in the aspects of its level of expression and speed of recovery.

Keywords

References

  1. Airaksinen S, Rabergh CM, Sistonen L, and Nikinmaa M (1998) Effects of heat shock and hypoxia on protein synthesis in rainbow trout (Oncorhynchus mykiss) cells. J Exp Biol 201:2543-2551.
  2. Bechtold DA, Rush SJ, Brown IR (2000) Localization of the heat-shock protein HSP70 to the synapse following hyperthermic stress in the brain. J Neurochem 74:641-646. https://doi.org/10.1046/j.1471-4159.2000.740641.x
  3. Currie S, Moyes CD, Tufts BL (2000) The effects of heat shock and acclimation temperature on hsp70 and hsp30 mRNA expression in rainbow trout: in vivo and in vitro comparisons. J Fish Biol 56:398-408. https://doi.org/10.1111/j.1095-8649.2000.tb02114.x
  4. Foster JA, Brown IR (1997) Differential induction of heat shock mRNA in oligodendrocytes, microglia, and astrocytes following hyperthermia. Mol Brain Res 45:207-218. https://doi.org/10.1016/S0169-328X(96)00138-6
  5. Iwama GK, Thomas PT, Forsyth RB, Vijayan MH (1998) Heat shock protein expression in fish. Rev Fish Biol Fisher 8:35-56. https://doi.org/10.1023/A:1008812500650
  6. Krone PH, Sass JB (1994) Hsp 90${\alpha}$ and Hsp 90${\beta}$ genes are present in the zebrafish and are differentially regulated in developing embryos. Biochem Biophys Res Comm 204:746-752. https://doi.org/10.1006/bbrc.1994.2522
  7. Krueger AM, Armstrong JN, Plumier J, Robertson HA, Currie RW (1999) Cell specific expression of Hsp70 in neurons and glia of the rat hippocampus after hyperthermia and kainic acid-induced seizure activity. Brain Res Mol Brain Res 71:265-278. https://doi.org/10.1016/S0169-328X(99)00198-9
  8. Lele Z, Engel S, Krone PH (1999) Hsp47 and hsp70 gene expression is differentially regulated in a stress- and tissue-specific manner in zebrafish embryos. Dev Genet 21:123-133. https://doi.org/10.1002/(SICI)1520-6408(1997)21:2<123::AID-DVG2>3.0.CO;2-9
  9. Li Y, Chopp M, Yoshida Y, Levine SR (1992) Distribution of 72-kDa heat-shock protein in rat brain after hyperthermia. Acta Neuropathol 84:94-99. https://doi.org/10.1007/BF00427221
  10. Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22:631-677. https://doi.org/10.1146/annurev.ge.22.120188.003215
  11. Maroni P, Bendinelli P, Tiberio L, Rovetta F, Piccoletti R, Schiaffonati L (2003) In vivo heat-shock response in the brain: signalling pathway and transcription factor activation. Brain Res Mol Brain Res 119:90-99. https://doi.org/10.1016/j.molbrainres.2003.08.018
  12. McCabe T, Simon RP (1993) Hyperthermia induces 72 kDa heat shock protein expression in rat brain in non-neuronal cells. Neurosci Lett 159:163-165. https://doi.org/10.1016/0304-3940(93)90824-5
  13. Menzerra P, Rush SJ, Brown IR (1997) Tissue-specific differences in heat shock protein hsc70 and hsp70 in the control and hyperthermic rabbit. J Cell Physiol 170:130-137. https://doi.org/10.1002/(SICI)1097-4652(199702)170:2<130::AID-JCP4>3.0.CO;2-P
  14. Morimoto RI, Kline MP, Bimston DN, Cotto JJ (1997) The heat-shock response: regulation and function of heat-shock proteins and molecular chaperones. Essays Biochem 32:17-29.
  15. Pearson DS, Kulyk WM, Kelly GM, Krone PH (1996) Cloning and characterization of a cDNA encoding the collagen-binding stress protein hsp47 in zebrafish. DNA Cell Biol 15:263-272. https://doi.org/10.1089/dna.1996.15.263
  16. Sass JB, Weinberg ES, Krone PH (1996) Specific localization of zebrafish hsp90${\alpha}$ mRNA tomyoD-expressing cells suggests a role for hsp90${\alpha}$ during normal muscle development. Mech Dev 54:195-204. https://doi.org/10.1016/0925-4773(95)00476-9
  17. Tsuchiya D, Hong S, Matsumori Y, Kayama T, Swanson RA, Dillman WH, Liu J, Panter SS, Weinstein PR (2003). Overexpression of rat heat shock protein 70 reduces neuronal injury after transient focal ischemia, transient global ischemia, or kainic acid-induced seizures. Neurosurgery 53:1179-1187. https://doi.org/10.1227/01.NEU.0000090341.38659.CF
  18. Walsh D, Li Z, Wu Y, Nagata K (1997) Heat shock and the role of the HSPs during neural plate induction in early mammalian CNS and brain development. Cell Mol Life Sci 53:198-211. https://doi.org/10.1007/PL00000592
  19. Westerfield M (2000) The Zebrafish Book. A Guide for the Laboratory Use of Zebrafish (Danio rerio). (4th ed.), University of Oregon Press, Eugene.
  20. Yenari MA, Liu J, Zheng Z, Vexler ZS, Lee JE, Giffard RG (2005) Antiapoptotic and anti-inflammatory mechanisms of heat-shock protein protection. Ann N Y Acad Sci 1053:74-83. https://doi.org/10.1196/annals.1344.007
  21. Zheng Z, Kim JY, Ma H, Lee JE, Yenari MA (2008). Anti-inflammatory effects of the 70 kDa heat shock protein in experimental stroke. J Cereb Blood Flow Metab 28:53-63. https://doi.org/10.1038/sj.jcbfm.9600502