Abstract
Krüppel-like factor 4 (KLF4) is a transcription factor with diverse and cell type-specific functions and is associated with a variety of pathophysiological processes. Recently, it has been proposed that the regulation of KLF4 is critical to neuronal differentiation and that neural progenitors overexpressing KLF4 take on a glial identity. The present study aimed to determine whether KLF4 is involved in the astroglial reaction induced by ischemia–reperfusion injury in the brain. No specific KLF4 immunoreactivity was observed in resting astrocytes of the control hippocampus, but significant induction was detected in reactive astrocytes preferentially located in the CA1 and dentate hilar regions of the hippocampus following transient forebrain ischemia. Astroglial KLF4 expression was induced in the nuclei and cytoplasm within 3 days of ischemia and persisted for at least 4 weeks. This pattern was reproduced in an in vitro astrogliosis model of rat primary cortical astrocytes exposed to oxygen–glucose deprivation (OGD). Furthermore, immunoblot assay showed that nuclear and cytosolic extracts from cortical astrocytes subjected to OGD had significantly higher levels of KLF4 protein compared to normoxic extracts. Thus, our data demonstrate that KLF4 expression was induced in astroglia by ischemic injury both in vivo and in vitro, suggesting that KLF4 may act as a transcription factor linked to the regulation of the astroglial reaction following ischemic injury.
Similar content being viewed by others
References
Acarin L, González B, Castellano B (2000) STAT3 and NFkB activation precedes glial reactivity in the excitotoxically injured young cortex but not in the corresponding distal thalamic nuclei. J Neuropathol Exp Neurol 59:151–163
Alder JK, Georgantas RW 3rd, Hildreth RL, Kaplan IM, Morisot S, Yu X, McDevitt M, Civin CI (2008) Kruppel-like factor 4 is essential for inflammatory monocyte differentiation in vivo. J Immunol 180:5645–5652
Behr R, Deller C, Godmann M, Müller T, Bergmann M, Ivell R, Steger K (2007) Kruppel-like factor 4 expression in normal and pathological human testes. Mol Hum Reprod 13:815–820
Chen Y, Swanson RA (2003) Astrocytes and brain injury. J Cereb Blood Flow Metab 23:137–149
Cho JM, Shoin YJ, Park JM, Kim J, Lee MY (2013) Characterization of nestin expression in astrocytes in the rat hippocampal CA1 region following transient forebrain ischemia. Anat Cell Biol 46:131–140
Choi JS, Kim SY, Cha JH, Choi YS, Sung KW, Oh ST, Kim ON, Chung JW, Chun MH, Lee SB, Lee MY (2003a) Upregulation of gp130 and STAT3 activation in the rat hippocampus following transient forebrain ischemia. Glia 41:237–246
Choi JS, Kim SY, Park HJ, Cha JH, Choi YS, Kang JE, Chung JW, Chun MH, Lee MY (2003b) Upregulation of gp130 and differential activation of STAT and p42/44 MAPK in the rat hippocampus following kainic acid-induced seizures. Mol Brain Res 119:10–18
Dang DT, Pevsner J, Yang VW (2000) The biology of the mammalian Krüppel-like family of transcription factors. Int J Biochem Cell Biol 32:1103–1121
de Pablo Y, Nilsson M, Pekna M, Pekny M (2013) Intermediate filaments are important for astrocyte response to oxidative stress induced by oxygen-glucose deprivation and reperfusion. Histochem Cell Biol 140:81–91
Dinapoli VA, Benkovic SA, Li X, Kelly KA, Miller DB, Rosen CL, Huber JD, O’Callaghan JP (2010) Age exaggerates proinflammatory cytokine signaling and truncates signal transducers and activators of transcription 3 signaling following ischemic stroke in the rat. Neuroscience 170:633–644
Feinberg MW, Cao Z, Wara AK, Lebedeva MA, Senbanerjee S, Jain MK (2005) Kruppel-like factor 4 is a mediator of proinflammatory signaling in macrophages. J Biol Chem 280:38247–38258
Garrett-Sinha LA, Eberspaecher H, Seldin MF, de Crombrugghe B (1996) A gene for a novel zinc-finger protein expressed in differentiated epithelial cells and transiently in certain mesenchymal cells. J Biol Chem 271:31384–31390
Hamik A, Lin Z, Kumar A, Balcells M, Sinha S, Katz J, Feinberg MW, Gerzsten RE, Edelman ER, Jain MK (2007) Kruppel-like factor 4 regulates endothelial inflammation. J Biol Chem 282:13769–13779
Herrmann JE, Imura T, Song B, Qi J, Ao Y, Nguyen TK, Korsak RA, Takeda K, Akira S, Sofroniew MV (2008) STAT3 is a critical regulator of astrogliosis and scar formation after spinal cord injury. J Neurosci 28:7231–7243
Jeong KH, Lee KE, Kim SY, Cho KO (2011) Upregulation of Krüppel-like factor 6 in the mouse hippocampus after pilocarpine-induced status epilepticus. Neuroscience 186:170–178
Jiang J, Chan YS, Loh YH, Cai J, Tong GQ, Lim CA, Robson P, Zhong S, Ng HH (2008) A core Klf circuitry regulates self-renewal of embryonic stem cells. Nat Cell Biol 10:353–360
Jorgensen MB, Finsen BR, Jensen MB, Castellano B, Diemer NH, Zimmer J (1993) Microglial and astroglial reactions to ischemic and kainic acid-induced lesions of the adult rat hippocampus. Exp Neurol 120:70–88
Justicia C, Gabriel C, Planas AM (2000) Activation of the Jak/Stat pathway following transient focal cerebral ischemia: signalling through Jak1 and Stat3 in astrocytes. Glia 30:253–270
Kaushik DK, Gupta M, Das S, Basu A (2010) Krüppel-like factor 4, a novel transcription factor regulates microglial activation and subsequent neuroinflammation. J Neuroinflammation 7:68
Kaushik DK, Mukhopadhyay R, Kumawat KL, Gupta M, Basu A (2012) Therapeutic targeting of Krüppel-like factor 4 abrogates microglial activation. J Neuroinflammation 9:57
Kirino T, Tamura A, Sano K (1985) Selective vulnerability of the hippocampus to ischemia—reversible and irreversible types of ischemic cell damage. Prog Brain Res 63:39–58
Le Magnen C, Bubendorf L, Ruiz C, Zlobec I, Bachmann A, Heberer M, Spagnoli GC, Wyler S, Mengus C (2013) Klf4 transcription factor is expressed in the cytoplasm of prostate cancer cells. Eur J Cancer 49:955–963
Lee MY, Shin SL, Choi YS, Kim EJ, Cha JH, Chun MH, Lee SB, Kim SY (1999) Transient upregulation of osteopontin mRNA in hippocampus and striatum following global forebrain ischemia in rats. Neurosci Lett 271:81–84
Li Y, McClintick J, Zhong L, Edenberg HJ, Yoder MC, Chan RJ (2005) Murine embryonic stem cell differentiation is promoted by SOCS-3 and inhibited by the zinc finger transcription factor Klf4. Blood 105:635–637
Li Q, Zhang R, Guo YL, Mei YW (2009) Effect of neuregulin on apoptosis and expressions of STAT3 and GFAP in rats following cerebral ischemic reperfusion. J Mol Neurosci 37:67–73
McConnell BB, Yang VW (2010) Mammalian Krüppel-like factors in health and diseases. Physiol Rev 90:1337–13381
Moore DL, Blackmore MG, Hu Y, Kaestner KH, Bixby JL, Lemmon VP, Goldberg JL (2009) KLF family members regulate intrinsic axon regeneration ability. Science 326:298–301
Moore DL, Apara A, Goldberg JL (2011) Krüppel-like transcription factors in the nervous system: novel players in neurite outgrowth and axon regeneration. Mol Cell Neurosci 47:233–243
Pekny M, Nilsson M (2005) Astrocyte activation and reactive gliosis. Glia 50:427–434
Pekny M, Pekna M (2004) Astrocyte intermediate filaments in CNS pathologies and regeneration. J Pathol 204:428–437
Pulsinelli WA, Brierley JB (1979) A new model of bilateral hemispheric ischemia in the unanesthetized rat. Stroke 10:267–272
Pulsinelli WA, Brierley JB, Plum F (1982) Temporal profile of neuronal damage in a model of transient forebrain ischemia. Ann Neurol 11:491–498
Qin S, Zhang CL (2012) Role of Kruppel-like factor 4 in neurogenesis and radial neuronal migration in the developing cerebral cortex. Mol Cell Biol 32:4297–4305
Qin S, Liu M, Niu W, Zhang CL (2011) Dysregulation of Kruppel-like factor 4 during brain development leads to hydrocephalus in mice. Proc Natl Acad Sci USA 108:21117–21121
Schmidt-Kastner R, Szymas J (1990) Immunohistochemistry of glial fibrillary acidic protein, vimentin and S-100 protein for study of astrocytes in hippocampus of rat. J Chem Neuroanat 3:179–192
Schubert KO, Naumann T, Schnell O, Zhi Q, Steup A, Hofmann HD, Kirsch M (2005) Activation of STAT3 signaling in axotomized neurons and reactive astrocytes after fimbria-fornix transaction. Exp Brain Res 165:520–531
Shen B, Smith RS Jr, Hsu YT, Chao L, Chao J (2009) Kruppel-like factor 4 is a novel mediator of Kallistatin in inhibiting endothelial inflammation via increased endothelial nitric-oxide synthase expression. J Biol Chem 284:35471–35478
Shie JL, Chen ZY, Fu M, Pestell RG, Tseng CC (2000) Gut-enriched Krüppel-like factor represses cyclin D1 promoter activity through Sp1 motif. Nucleic Acids Res 28:2969–2976
Shields JM, Christy RJ, Yang VW (1996) Identification and characterization of a gene encoding a gut-enriched Krüppel-like factor expressed during growth arrest. J Biol Chem 271:20009–20017
Suzuki S, Tanaka K, Nogawa S, Dembo T, Kosakai A, Fukuuchi Y (2001) Phosphorylation of signal transducer and activator of transcription-3 (Stat3) after focal cerebral ischemia in rats. Exp Neurol 170:63–71
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663–676
Wang R, Zahng X, Zhang J, Fan Y, Shen Y, Hu W, Chen Z (2012) Oxygen-glucose deprivation induced glial scar-like change in astrocytes. PLoS ONE 7:e37574
Wei D, Wang L, Kanai M, Jia Z, Le X, Li Q, Wang H, Xie K (2010) KLF4α up-regulation promotes cell cycle progression and reduces survival time of patients with pancreatic cancer. Gastroenterology 139:2135–2145
Xu Z, Xue T, Zhang Z, Wang X, Xu P, Zhang J, Lei X, Li Y, Xie Y, Wang L, Fang M, Chen Y (2011) Role of signal transducer and activator of transcription-3 in up-regulation of GFAP after epilepsy. Neurochem Res 36:2208–2215
Zamanian JL, Xu L, Foo LC, Nouri N, Zhou L, Giffard RG, Barres BA (2012) Genomic analysis of reactive astrogliosis. J Neurosci 32:6391–6410
Zhang W, Geiman DE, Shields JM, Dang DT, Mahatan CS, Kaestner KH, Biggs JR, Kraft AS, Yang VW (2000) The Gut-enriched Kruppel-like factor (Kruppel-like factor 4) mediates the transactivating effect of p53 on the p21WAF1/Cip1 promoter. J Biol Chem 275:18391–18398
Acknowledgments
This study was supported by the Mid-career Researcher Program through the National Research Foundation of Korea (NRF) grant funded by the MEST (2011-0028319).
Author information
Authors and Affiliations
Corresponding author
Additional information
Joo-Hee Park and Tae-Ryong Riew have contributed equally to this study.
Rights and permissions
About this article
Cite this article
Park, JH., Riew, TR., Shin, YJ. et al. Induction of Krüppel-like factor 4 expression in reactive astrocytes following ischemic injury in vitro and in vivo. Histochem Cell Biol 141, 33–42 (2014). https://doi.org/10.1007/s00418-013-1134-5
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-013-1134-5