Overexpression of netrin‑1 increases the expression of tight junction‑associated proteins, claudin‑5, occludin, and ZO‑1, following traumatic brain injury in rats

Wen,Jianfeng; Qian,Suokai; Yang,Qifan; Deng,Lei; Mo,Ye; Yu,Yuefei

doi:10.3892/etm.2014.1818

Overexpression of netrin‑1 increases the expression of tight junction‑associated proteins, claudin‑5, occludin, and ZO‑1, following traumatic brain injury in rats

Authors:
- Jianfeng Wen
- Suokai Qian
- Qifan Yang
- Lei Deng
- Ye Mo
- Yuefei Yu
View Affiliations

Affiliations: Department of Neurosurgery, Changcheng Sub‑Hospital of Nanchang University, Nanchang, Jiangxi 330002, P.R. China, Department of Neurosurgery, The 94th Hospital of PLA, Nanchang, Jiangxi 330002, P.R. China, Department of Nursing, The 94th Hospital of PLA, Nanchang, Jiangxi 330002, P.R. China
Published online on: July 1, 2014 https://doi.org/10.3892/etm.2014.1818
Pages: 881-886

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The function of the blood‑brain barrier (BBB) depends on the integrity of tight junction (TJ)‑associated proteins. Netrin‑1 is known to promote angiogenesis and may also regulate the BBB. To understand the association between netrin‑1 and the TJ‑associated proteins, the expression levels of proteins involved in maintaining the integrity of the BBB, including netrin‑1, claudin‑5, occludin and zonula occluden (ZO)‑1, were investigated in the present study using quantitative polymerase chain reaction, western blot analysis and immunofluorescence. The aim of the present study was to determine the changes in BBB permeability and whether pZsGreen1‑N1 mediated overexpression of netrin‑1 increased the expression of the TJ‑associated proteins following traumatic brain injury (TBI). The results demonstrated that the levels of mRNA transcription and protein expression of the TJ‑associated proteins, claudin‑5, occludin and ZO‑1, were significantly reduced following TBI. Furthermore, the changes in the expression of these three TJ proteins were consistent with the changes in the BBB permeability, indicating that weakening intercellular junctions leads to BBB opening. The present study also demonstrated that netrin‑1 significantly increased the downregulation of claudin‑5, occludin and ZO‑1 expression levels induced by TBI, which provided a basis for further investigation on the role of netrin‑1 in the integrity of TJs and proper functioning of the BBB.

View Figures

View References

1	Fanning AS, Mitic LL and Anderson JM: Transmembrane proteins in the tight junction barrier. J Am Soc Nephrol. 10:1337–1345. 1999.PubMed/NCBI
2	Furuse M, Sasaki H and Tsukita S: Manner of interaction of heterogeneous claudin species within and between tight junction strands. J Cell Biol. 147:891–903. 1999. View Article : Google Scholar : PubMed/NCBI
3	Hirase T, Staddon JM, Saitou M, et al: Occludin as a possible determinant of tight junction permeability in endothelial cells. J Cell Sci. 110:1603–1613. 1997.PubMed/NCBI
4	Anderson J, Fanning A, Lapierre L and Van Itallie CM: Zonula occludens (ZO)-1 and ZO-2: membrane-associated guanylate kinase homologues (MAGuKs) of the tight junction. Biochem Soc Trans. 23:470–475. 1995.PubMed/NCBI
5	Haskins J, Gu L, Wittchen ES, Hibbard J and Stevenson BR: ZO-3, a novel member of the MAGUK protein family found at the tight junction, interacts with ZO-1 and occludin. J Cell Biol. 141:199–208. 1998. View Article : Google Scholar : PubMed/NCBI
6	Huber JD, Egleton RD and Davis TP: Molecular physiology and pathophysiology of tight junctions in the blood-brain barrier. Trends Neurosci. 24:719–725. 2001. View Article : Google Scholar : PubMed/NCBI
7	Park KW, Crouse D, Lee M, et al: The axonal attractant netrin-1 is an angiogenic factor. Proc Natl Acad Sci USA. 101:16210–16215. 2004. View Article : Google Scholar : PubMed/NCBI
8	Wilson BD, Ii M, Park KW, et al: Netrins promote developmental and therapeutic angiogenesis. Science. 313:640–644. 2006. View Article : Google Scholar : PubMed/NCBI
9	Shapira Y, Setton D, Artru AA and Shohami E: Blood-brain barrier permeability, cerebral edema, and neurologic function after closed head injury in rats. Anesth Analg. 77:141–148. 1993. View Article : Google Scholar : PubMed/NCBI
10	Soares HD, Thomas M, Cloherty K and McIntosh TK: Development of prolonged focal cerebral edema and regional cation changes following experimental brain injury in the rat. J Neurochem. 58:1845–1852. 1992. View Article : Google Scholar : PubMed/NCBI
11	Feeney DM, Boyeson MG, Linn RT, Murray HM and Dail WG: Responses to cortical injury: I. Methodology and local effects of contusions in the rat. Brain Res. 211:67–77. 1981. View Article : Google Scholar : PubMed/NCBI
12	Belayev L, Busto R, Zhao W and Ginsberg MD: Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats. Brain Res. 739:88–96. 1996. View Article : Google Scholar : PubMed/NCBI
13	Zeng Y, Liu Z, Yang J, et al: ARID1A is a tumour suppressor and inhibits glioma cell proliferation via the PI3K pathway. Head Neck Oncol. 5:62013.
14	Bamforth SD, Kniesel U, Wolburg H, Engelhardt B and Risau W: A dominant mutant of occludin disrupts tight junction structure and function. J Cell Sci. 112:1879–1888. 1999.PubMed/NCBI
15	Feldman GJ, Mullin JM and Ryan MP: Occludin: structure, function and regulation. Adv Drug Deliv Rev. 57:883–917. 2005. View Article : Google Scholar : PubMed/NCBI
16	Piontek J, Winkler L, Wolburg H, et al: Formation of tight junction: determinants of homophilic interaction between classic claudins. FASEB J. 22:146–158. 2008. View Article : Google Scholar : PubMed/NCBI
17	Nitta T, Hata M, Gotoh S, et al: Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice. J Cell Biol. 161:653–660. 2003. View Article : Google Scholar : PubMed/NCBI
18	Honda M, Nakagawa S, Hayashi K, Kitagawa N, et al: Adrenomedullin improves the blood-brain barrier function through the expression of claudin-5. Cell Mol Neurobiol. 26:109–118. 2006. View Article : Google Scholar : PubMed/NCBI
19	Umeda K, Ikenouchi J, Katahira-Tayama S, et al: ZO-1 and ZO-2 independently determine where claudins are polymerized in tight-junction strand formation. Cell. 126:741–754. 2006. View Article : Google Scholar : PubMed/NCBI
20	Umeda K, Matsui T, Nakayama M, et al: Establishment and characterization of cultured epithelial cells lacking expression of ZO-1. J Biol Chem. 279:44785–44794. 2004. View Article : Google Scholar : PubMed/NCBI
21	Fan Y, Shen F, Chen Y, et al: Overexpression of netrin-1 induces neovascularization in the adult mouse brain. J Cereb Blood Flow Metab. 28:1543–1551. 2008. View Article : Google Scholar : PubMed/NCBI
22	Liu N, Huang H, Lin F, Chen A, Zhang Y, Chen R and Du H: Effects of treadmill exercise on the expression of netrin-1 and its receptors in rat brain after cerebral ischemia. Neuroscience. 194:349–358. 2011. View Article : Google Scholar : PubMed/NCBI