Overexpression of Slit2 improves function of the paravascular pathway in the aging mouse brain

Li,Ge; He,Xiaofei; Li,Hang; Wu,Yu'e; Guan,Yalun; Liu,Shuhua; Jia,Huanhuan; Li,Yunfeng; Wang,Lijing; Huang,Ren; Pei,Zhong; Lan,Yue; Zhang,Yu

doi:10.3892/ijmm.2018.3802

Overexpression of Slit2 improves function of the paravascular pathway in the aging mouse brain

Authors:
- Ge Li
- Xiaofei He
- Hang Li
- Yu'e Wu
- Yalun Guan
- Shuhua Liu
- Huanhuan Jia
- Yunfeng Li
- Lijing Wang
- Ren Huang
- Zhong Pei
- Yue Lan
- Yu Zhang
View Affiliations

Affiliations: Guangdong Provincial Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, Guangdong 510663, P.R. China, Department of Neurology, National Key Clinical Department and Key Discipline of Neurology, Guangdong Key Laboratory for Diagnosis and Treatment of Major Neurological Diseases, The First Affiliated Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China, Vascular Biology Research Institute, School of Basic Course, Guangdong Pharmaceutical University, Guangzhou, Guangdong 510006, P.R. China, Department of Rehabilitation Medicine, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510180, P.R. China
Published online on: August 2, 2018 https://doi.org/10.3892/ijmm.2018.3802
Pages: 1935-1944
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Aging is associated with impairment of the paravascular pathway caused by the activation of astrocytes and depolarization of protein aquaporin‑4 (AQP4) water channels, resulting in the accumulation of protein waste, including amyloid β (Aβ), in the brain parenchyma. The secreted glycoprotein slit guidance ligand 2 (Slit2) is important in regulating the function of the central nervous system and inflammatory response process. In the present study, 15‑month‑old Slit2 overexpression transgenic mice (Slit2‑Tg mice) and two‑photon fluorescence microscopy were used to evaluate the dynamic clearance of the paravascular pathway and the integrity of the blood‑brain barrier (BBB). The reactivity of astrocytes, polarity of AQP4 and deposition of Aβ in the brain parenchyma were analyzed by immunofluorescence. A Morris water maze test was used to examine the effect of Slit2 on spatial memory cognition in aging mice. It was found that the overexpression of Slit2 improved the clearance of the paravascular pathway by inhibiting astrocyte activation and maintaining AQP4 polarity on the astrocytic endfeet in Slit2‑Tg mice. In addition, Slit2 restored the disruption of the BBB caused by aging. The accumulation of Aβ was significantly reduced in the brain of Slit2‑Tg mice. Furthermore, the water maze experiment showed that Slit2 improved spatial memory cognition in the aging mice. These results indicated that Slit2 may have the potential to be used in the prevention and treatment of neurodegenerative diseases in the elderly.

View Figures

View References

1	Ross CA and Poirier MA: Protein aggregation and neurodegenerative disease. Nat Med. 10(Suppl): S10–S17. 2004. View Article : Google Scholar : PubMed/NCBI
2	Iliff JJ, Wang M, Liao Y, Plogg BA, Peng W, Gundersen GA, Benveniste H, Vates GE, Deane R, Goldman SA, et al: A paravascular pathway facilitates CSF flow through the brain parenchyma and the clearance of interstitial solutes, including amyloid beta. Sci Transl Med. 4:147ra1112012. View Article : Google Scholar
3	Kress BT, Iliff JJ, Xia M, Wang M, Wei HS, Zeppenfeld D, Xie L, Kang H, Xu Q, Liew JA, et al: Impairment of paravascular clearance pathways in the aging brain. Ann Neurol. 76:845–861. 2014. View Article : Google Scholar : PubMed/NCBI
4	Iliff JJ, Wang M, Zeppenfeld DM, Venkataraman A, Plog BA, Liao Y, Deane R and Nedergaard M: Cerebral arterial pulsation drives paravascular CSF-interstitial fluid exchange in the murine brain. J Neurosci. 33:18190–18199. 2013. View Article : Google Scholar : PubMed/NCBI
5	Iliff JJ, Lee H, Yu M, Feng T, Logan J, Nedergaard M and Benveniste H: Brain-wide pathway for waste clearance captured by contrast-enhanced MRI. J Clin Invest. 123:1299–1309. 2013. View Article : Google Scholar : PubMed/NCBI
6	Brose K, Bland KS, Wang KH, Arnott D, Henzel W, Goodman CS, Tessier-Lavigne M and Kidd T: Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell. 96:795–806. 1999. View Article : Google Scholar : PubMed/NCBI
7	Ypsilanti AR, Zagar Y and Chedotal A: Moving away from the midline: New developments for Slit and Robo. Development. 137:1939–1952. 2010. View Article : Google Scholar : PubMed/NCBI
8	Altay T, McLaughlin B, Wu JY, Park TS and Gidday JM: Slit modulates cerebrovascular inflammation and mediates neuroprotection against global cerebral ischemia. Exp Neurol. 207:186–194. 2007. View Article : Google Scholar : PubMed/NCBI
9	Park JH, Pak HJ, Riew TR, Shin YJ and Lee MY: Increased expression of Slit2 and its receptors Robo1 and Robo4 in reactive astrocytes of the rat hippocampus after transient forebrain ischemia. Brain Res. 1634:45–56. 2016. View Article : Google Scholar : PubMed/NCBI
10	Han HX and Geng JG: Over-expression of Slit2 induces vessel formation and changes blood vessel permeability in mouse brain. Acta Pharmacol Sin. 32:1327–1336. 2011. View Article : Google Scholar : PubMed/NCBI
11	Yuen DA and Robinson LA: Slit2-Robo signaling: A novel regulator of vascular injury. Curr Opin Nephrol Hypertens. 22:445–451. 2013. View Article : Google Scholar : PubMed/NCBI
12	Elahy M, Jackaman C, Mamo JC, Lam V, Dhaliwal SS, Giles C, Nelson D and Takechi R: Blood-brain barrier dysfunction developed during normal aging is associated with inflammation and loss of tight junctions but not with leukocyte recruitment. Immun Ageing. 12:22015. View Article : Google Scholar : PubMed/NCBI
13	Gorlé N, Van Cauwenberghe C, Libert C and Vandenbroucke RE: The effect of aging on brain barriers and the consequences for Alzheimer's disease development. Mamm Genome. 27:407–420. 2016. View Article : Google Scholar : PubMed/NCBI
14	National Research Council (U.S.), Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research (U.S.) and National Academies Press (U.S.): Guide for the Care and Use of Laboratory Animals. National Academies Press; Washington, D.C.: 2011
15	Li JC, Han L, Wen YX, Yang YX, Li S, Li XS, Zhao CJ, Wang TY, Chen H, Liu Y, et al: Increased permeability of the blood-brain barrier and Alzheimer's disease-like alterations in slit-2 transgenic mice. J Alzheimers Dis. 43:535–548. 2015. View Article : Google Scholar
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	He XF, Lan Y, Zhang Q, Liu DX, Wang Q, Liang FY, Zeng JS, Xu GQ and Pei Z: Deferoxamine inhibits microglial activation, attenuates blood-brain barrier disruption, rescues dendritic damage and improves spatial memory in a mouse model of microhemorrhages. J Neurochem. 138:436–447. 2016. View Article : Google Scholar : PubMed/NCBI
18	Luo C, Yao X, Li J, He B, Liu Q, Ren H, Liang F, Li M, Lin H, Peng J, et al: Paravascular pathways contribute to vasculitis and neuroinflammation after subarachnoid hemorrhage independently of glymphatic control. Cell Death Dis. 7:e21602016. View Article : Google Scholar : PubMed/NCBI
19	Harvey CD, Coen P and Tank DW: Choice-specific sequences in parietal cortex during a virtual-navigation decision task. Nature. 484:62–68. 2012. View Article : Google Scholar : PubMed/NCBI
20	Ren Z, Iliff JJ, Yang L, Yang J, Chen X, Chen MJ, Giese RN, Wang B, Shi X and Nedergaard M: 'Hit & Run' model of closed-skull traumatic brain injury (TBI) reveals complex patterns of post-traumatic AQP4 dysregulation. J Cereb Blood Flow Metab. 33:834–845. 2013. View Article : Google Scholar : PubMed/NCBI
21	Wang M, Iliff JJ, Liao Y, Chen MJ, Shinseki MS, Venkataraman A, Cheung J, Wang W and Nedergaard M: Cognitive deficits and delayed neuronal loss in a mouse model of multiple microinfarcts. J Neurosci. 32:17948–17960. 2012. View Article : Google Scholar : PubMed/NCBI
22	Wu H, Wu T, Li M and Wang J: Efficacy of the lipid-soluble iron chelator 2,2′-dipyridyl against hemorrhagic brain injury. Neurobiol Dis. 45:388–394. 2012. View Article : Google Scholar
23	Di Marco LY, Farkas E, Martin C, Venneri A and Frangi AF: Is vasomotion in cerebral arteries impaired in Alzheimer's disease? J Alzheimers Dis. 46:35–53. 2015. View Article : Google Scholar : PubMed/NCBI
24	Nhan T, Burgess A, Cho EE, Stefanovic B, Lilge L and Hynynen K: Drug delivery to the brain by focused ultrasound induced blood-brain barrier disruption: Quantitative evaluation of enhanced permeability of cerebral vasculature using two-photon microscopy. J Control Release. 172:274–280. 2013. View Article : Google Scholar : PubMed/NCBI
25	Hladky SB and Barrand MA: Mechanisms of fluid movement into, through and out of the brain: Evaluation of the evidence. Fluids Barriers CNS. 11:262014. View Article : Google Scholar
26	Bakker EN, Bacskai BJ, Arbel-Ornath M, Aldea R, Bedussi B, Morris AW, Weller RO and Carare RO: Lymphatic Clearance of the Brain: Perivascular, paravascular and significance for neurodegenerative diseases. Cell Mol Neurobiol. 36:181–194. 2016. View Article : Google Scholar : PubMed/NCBI
27	Iliff JJ, Chen MJ, Plog BA, Zeppenfeld DM, Soltero M, Yang L, Singh I, Deane R and Nedergaard M: Impairment of glymphatic pathway function promotes tau pathology after traumatic brain injury. J Neurosci. 34:16180–16193. 2014. View Article : Google Scholar : PubMed/NCBI
28	Verkman AS, Anderson MO and Papadopoulos MC: Aquaporins: Important but elusive drug targets. Nat Rev Drug Discov. 13:259–277. 2014. View Article : Google Scholar : PubMed/NCBI
29	Blockus H and Chedotal A: Slit-Robo signaling. Development. 143:3037–3044. 2016. View Article : Google Scholar : PubMed/NCBI
30	London NR and Li DY: Robo4-dependent Slit signaling stabilizes the vasculature during pathologic angiogenesis and cytokine storm. Curr Opin Hematol. 18:186–190. 2011. View Article : Google Scholar : PubMed/NCBI
31	Zhao H, Anand AR and Ganju RK: Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia. J Immunol. 192:385–393. 2014. View Article : Google Scholar
32	Pekny M, Pekna M, Messing A, Steinhäuser C, Lee JM, Parpura V, Hol EM, Sofroniew MV and Verkhratsky A: Astrocytes: A central element in neurological diseases. Acta Neuropathol. 131:323–345. 2016. View Article : Google Scholar
33	Farina C, Aloisi F and Meinl E: Astrocytes are active players in cerebral innate immunity. Trends Immunol. 28:138–145. 2007. View Article : Google Scholar : PubMed/NCBI
34	Hagino S, Iseki K, Mori T, Zhang Y, Hikake T, Yokoya S, Takeuchi M, Hasimoto H, Kikuchi S and Wanaka A: Slit and glypican-1 mRNAs are coexpressed in the reactive astrocytes of the injured adult brain. Glia. 42:130–138. 2003. View Article : Google Scholar : PubMed/NCBI
35	Sherchan P, Huang L, Wang Y, Akyol O, Tang J and Zhang JH: Recombinant Slit2 attenuates neuroinflammation after surgical brain injury by inhibiting peripheral immune cell infiltration via Robo1-srGAP1 pathway in a rat model. Neurobiol Dis. 85:164–173. 2016. View Article : Google Scholar
36	Jones CA, London NR, Chen H, Park KW, Sauvaget D, Stockton RA, Wythe JD, Suh W, Larrieu-Lahargue F, Mukouyama YS, et al: Robo4 stabilizes the vascular network by inhibiting pathologic angiogenesis and endothelial hyperpermeability. Nat Med. 14:448–453. 2008. View Article : Google Scholar : PubMed/NCBI
37	Jones CA, Nishiya N, London NR, Zhu W, Sorensen LK, Chan AC, Lim CJ, Chen H, Zhang Q, Schultz PG, et al: Slit2-Robo4 signalling promotes vascular stability by blocking Arf6 activity. Nat Cell Biol. 11:1325–1331. 2009. View Article : Google Scholar : PubMed/NCBI
38	Moody DM, Brown WR, Challa VR, Ghazi-Birry HS and Reboussin DM: Cerebral microvascular alterations in aging, leukoaraiosis, and Alzheimer's disease. Ann NY Acad Sci. 826:103–116. 1997. View Article : Google Scholar : PubMed/NCBI
39	Saunders NR, Dziegielewska KM, Møllgård K and Habgood MD: Markers for blood-brain barrier integrity: How appropriate is Evans blue in the twenty-first century and what are the alternatives? Front Neurosci. 9:3852015. View Article : Google Scholar : PubMed/NCBI
40	Deacon RMJ: APP-based transgenic models: The Tg2576 model. Animal Models of Dementia. De Deyn PP and Van Dam D: Humana Press; Totowa, NJ: pp. 387–398. 2011, View Article : Google Scholar
41	Yang W, Wu Q, Yuan C, Gao J, Xiao M, Gu M, Ding J and Hu G: Aquaporin-4 mediates astrocyte response to beta-amyloid. Mol Cell Neurosci. 49:406–414. 2012. View Article : Google Scholar : PubMed/NCBI
42	Wilcock DM, Vitek MP and Colton CA: Vascular amyloid alters astrocytic water and potassium channels in mouse models and humans with Alzheimer's disease. Neuroscience. 159:1055–1069. 2009. View Article : Google Scholar : PubMed/NCBI