Abstract
Juxtanodin (JN) is a cytoskeleton-related oligodendrocyte-specific gene, and its underlying mechanism still needs detailed exploration. In this study, we tested whether a 1.9 kb fragment from the 5′-flanking region of JN is sufficient to specifically deliver the gene expression in oligodendrocytes. By reporter assay, we found that the 1.9 kb fragment specifically activated in C6 cells, which is further upregulated by ATRA-induced oligodendrocyte lineage differentiation. Bioinformatics study revealed that Nkx2.2 might be a transcription factor involved in the process. qPCR results showed that ATRA upregulated endogenous expression of Nkx2.2 in C6 cells. Further study revealed that Nkx2.2 could bind JN promoter and its overexpression increase the promoter activity of JN. In summary, our study here suggests that Nkx2.2 is one of the essential transcription factors delivering the specificity of JN promoter in oligodendrocytes.
Similar content being viewed by others
References
Bradl M, Lassmann H (2009) Oligodendrocytes: biology and pathology. Acta Neuropathol 119(1):37–53
Fulton D, Paez PM, Campagnoni AT (2009) The multiple roles of myelin protein genes during the development of the oligodendrocyte. ASN Neuro 2(1):e00027
Zhang B, Cao Q, Guo A, Chu H, Chan YG, Buschdorf JP, Low BC, Ling EA, Liang F (2005) Juxtanodin: an oligodendroglial protein that promotes cellular arborization and 2′, 3′-cyclic nucleotide-3′-phosphodiesterase trafficking. Proc Natl Acad Sci USA 102:11527–11532
Qi Y, Cai J, Wu Y, Wu R, Lee J, Fu H, Rao M, Sussel L, Rubenstein J, Qiu M (2001) Control of oligodendrocyte differentiation by the Nkx2.2 homeodomain transcription factor. Development 128:2723–2733
Soula C, Danesin C, Kan P, Grob M, Poncet C, Cochard P (2001) Distinct sites of origin of oligodendrocytes and somatic motoneurons in the chick spinal cord: oligodendrocytes arise from Nkx2.2-expressing progenitors by a Shh-dependent mechanism. Development 128:1369–1379
Fancy SP, Zhao C, Franklin RJ (2004) Increased expression of Nkx2.2 and Olig2 identifies reactive oligodendrocyte progenitor cells responding to demyelination in the adult CNS. Mol Cell Neurosci 27:247–254
Fu H, Qi Y, Tan M, Cai J, Takebayashi H, Nakafuku M, Richardson W, Qiu M (2002) Dual origin of spinal oligodendrocyte progenitors and evidence for the cooperative role of Olig2 and Nkx2.2 in the control of oligodendrocyte differentiation. Development 129:681–693
Yang G, Yu F, Fu H, Lu F, Huang B, Bai L, Zhao Z, Yao L, Lu Z (2007) Identification of the distinct promoters for the two transcripts of apoptosis related protein 3 and their transcriptional regulation by NFAT and NFkappaB. Mol Cell Biochem 302:187–194
Fischer I, Nolan CE, Shea TB (1987) Effects of retinoic acid on expression of the transformed phenotype in C6 glioma cells. Life Sci 41:463–470
Zhu W, Kanoh M, Ye P, Laszkiewicz I, Royland JE, Wiggins RC, Konat G (1992) Retinoic acid-regulated expression of proteolipid protein and myelin-associated glycoprotein genes in C6 glioma cells. J Neurosci Res 31:745–750
Lopez-Barahona M, Minano M, Mira E, Iglesias T, Stunnenberg HG, Rodriguez-Pena A, Bernal J, Munoz A (1993) Retinoic acid posttranscriptionally up-regulates proteolipid protein gene expression in C6 glioma cells. J Biol Chem 268:25617–25623
Bianchi MG, Gazzola GC, Tognazzi L, Bussolati O (2008) C6 glioma cells differentiated by retinoic acid overexpress the glutamate transporter excitatory amino acid carrier 1 (EAAC1). Neuroscience 151:1042–1052
Kursula P (2001) The current status of structural studies on proteins of the myelin sheath (review). Int J Mol Med 8:475–479
Kursula P (2008) Structural properties of proteins specific to the myelin sheath. Amino Acids 34:175–185
Haines JD, Fragoso G, Hossain S, Mushynski WE, Almazan G (2008) p38 Mitogen-activated protein kinase regulates myelination. J Mol Neurosci 35:23–33
Kamholz JA (1996) Regulation of myelin development. Mult Scler 2:236–240
Kamholz J, Awatramani R, Menichella D, Jiang H, Xu W, Shy M (1999) Regulation of myelin-specific gene expression. Relevance to CMT1. Ann NY Acad Sci 883:91–108
Wei Q, Miskimins WK, Miskimins R (2005) Stage-specific expression of myelin basic protein in oligodendrocytes involves Nkx2.2-mediated repression that is relieved by the Sp1 transcription factor. J Biol Chem 280:16284–16294
Calatayud CA, Garcia CI, Paez PM, Pasquini JM, Soto EF, Pasquini LA (2009) Partial inhibition of the proteasome enhances the activity of the myelin basic protein promoter. Dev Neurosci 31:169–180
Zheng X, Shen G, Yang X, Liu W (2007) Most C6 cells are cancer stem cells: evidence from clonal and population analyses. Cancer Res 67:3691–3697
Acknowledgments
This work was supported by research grants from the National Natural Science Foundation of China (No. 30770420, No. 30871239).
Author information
Authors and Affiliations
Corresponding authors
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wang, T., Jia, L., Yang, G. et al. Identification of Juxtanodin promoter and its transcriptional regulation during the ATRA-induced differentiation of C6 cells. Mol Cell Biochem 350, 177–183 (2011). https://doi.org/10.1007/s11010-010-0696-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-010-0696-y