Abstract
The choroid plexus epithelium forms the interface between the blood and the cerebrospinal fluid. In addition to its barrier function resulting from the presence of tight junctions sealing the epithelial cells together, the choroid plexus epithelium fulfills vectorial transport (influx and efflux), neuroprotective, antioxidant and secretory functions, all relevant to different aspects of neurotoxicological sciences. To investigate these choroidal functions without the interference of the blood–brain barrier proper and brain parenchyma, in vitro cellular models of the blood–cerebrospinal fluid barrier, retaining the differentiated phenotype of the choroidal epithelium, have been established, taking advantage of the advent of refined culture methods and availability of permeable membranes. This chapter provides information to help investigators to set up and characterize choroid plexus epithelial cells in culture in bicameral devices. It first describes the factors that are critical to isolate the cells and select the culture conditions, and provides a survey of available cell lines with their advantages and limitations. Then the primordial specific choroidal features that can be examined within a validation scheme are discussed, emphasizing the need for a careful interpretation of their significance. In a third part, selected examples of studies performed with these models are presented, highlighting their potential applications in the field of neurotoxicology.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Davson H, Segal MB (1996) Physiology of the CSF and the blood–brain barriers. CRC Press, Boca Raton
Strazielle N, Ghersi-Egea JF (2000) Choroid plexus in the central nervous system biology and physiopathology. J Neuropathol Exp Neurol 59:561–574
Johansson PA, Dziegielewska KM, Liddelow SA, Saunders NR (2008) The blood–CSF barrier explained: when development is not immaturity. Bioessays 30:237–248
Charo IF, Ransohoff RM (2006) The many roles of chemokines and chemokine receptors in inflammation. N Engl J Med 354:610–621
Szmydynger-Chodobska J, Strazielle N, Zink BJ, Ghersi-Egea JF, Chodobski A (2009) The role of the choroid plexus in neutrophil invasion after traumatic brain injury. J Cereb Blood Flow Metab 27:1503–1516
Strazielle N, Preston JE (2003) Transport across the choroid plexuses in vivo and in vitro. Methods Mol Med 89:291–304
Crook RB, Kasagami H, Prusiner SB (1981) Culture and characterization of epithelial cells from bovine choroid plexus. J Neurochem 37:845–854
Holm NR, Hansen LB, Nilsson C, Gammeltoft S (1994) Gene expression and secretion of insulin-like growth factor-II and insulin-like growth factor binding protein-2 from cultured sheep choroid plexus epithelial cells. Brain Res Mol Brain Res 21:67–74
Mayer SE, Sanders-Bush E (1993) Sodium-dependent antiporters in choroid plexus epithelial cultures from rabbit. J Neurochem 60:1308–1316
Plotkin MD, Kaplan MR, Peterson LN, Gullans SR, Hebert SC, Delpire E (1997) Expression of the Na(+)-K(+)-2Cl-cotransporter BSC2 in the nervous system. Am J Physiol 272:C173–C183
Shu C, Shen H, Teuscher NS, Lorenzi PJ, Keep RF, Smith DE (2002) Role of PEPT2 in peptide/mimetic trafficking at the blood–cerebrospinal fluid barrier: studies in rat choroid plexus epithelial cells in primary culture. J Pharmacol Exp Ther 301:820–829
Strazielle N, Ghersi-Egea JF (1999) Demonstration of a coupled metabolism-efflux process at the choroid plexus as a mechanism of brain protection toward xenobiotics. J Neurosci 19:6275–6289
Villalobos AR, Parmelee JT, Pritchard JB (1997) Functional characterization of choroid plexus epithelial cells in primary culture. J Pharmacol Exp Ther 282:1109–1116
Zheng W, Zhao Q, Graziano JH (1998) Primary culture of choroidal epithelial cells: characterization of an in vitro model of blood–CSF barrier. In Vitro Cell Dev Biol Anim 34:40–45
Tsutsumi M, Skinner MK, Sanders-Bush E (1989) Transferrin gene expression and synthesis by cultured choroid plexus epithelial cells. Regulation by serotonin and cyclic adenosine 3′, 5′-monophosphate. J Biol Chem 264:9626–9631
Gath U, Hakvoort A, Wegener J, Decker S, Galla HJ (1997) Porcine choroid plexus cells in culture: expression of polarized phenotype, maintenance of barrier properties and apical secretion of CSF-components. Eur J Cell Biol 74:68–78
Ramanathan VK, Hui AC, Brett CM, Giacomini KM (1996) Primary cell culture of the rabbit choroid plexus: an experimental system to investigate membrane transport. Pharm Res 13:952–956
Southwell BR, Duan W, Alcorn D, Brack C, Richardson SJ, Kohrle J, Schreiber G (1993) Thyroxine transport to the brain: role of protein synthesis by the choroid plexus. Endocrinology 133:2116–2126
Haselbach M, Wegener J, Decker S, Engelbertz C, Galla HJ (2001) Porcine choroid plexus epithelial cells in culture: regulation of barrier properties and transport processes. Microsc Res Tech 52:137–152
Sanders-Bush E, Breeding M (1991) Choroid plexus epithelial cells in primary culture: a model of 5HT1C receptor activation by hallucinogenic drugs. Psychopharmacology (Berl) 105:340–346
Hoffmann A, Gath U, Gross G, Lauber J, Getzlaff R, Hellwig S, Galla HJ, Conradt HS (1996) Constitutive secretion of beta-trace protein by cultivated porcine choroid plexus epithelial cells: elucidation of its complete amino acid and cDNA sequences. J Cell Physiol 169:235–241
Strazielle N, Khuth ST, Murat A, Chalon A, Giraudon P, Belin MF, Ghersi-Egea JF (2003) Pro-inflammatory cytokines modulate matrix metalloproteinase secretion and organic anion transport at the blood–cerebrospinal fluid barrier. J Neuropathol Exp Neurol 62:1254–1264
Hakvoort A, Haselbach M, Wegener J, Hoheisel D, Galla HJ (1998) The polarity of choroid plexus epithelial cells in vitro is improved in serum-free medium. J Neurochem 71:1141–1150
Strazielle N, Ghersi-Egea JF (2005) In vitro investigation of the blood–cerebrospinal fluid barrier properties: primary cultures and immortalized cell lines of the choroidal epithelium. In: Zheng W, Chodobski A (eds) The blood–cerebrospinal fluid barrier. Taylor and Francis, Boca Raton, pp 553–593
Nilsson C, Hultberg BM, Gammeltoft S (1996) Autocrine role of insulin-like growth factor II secretion by the rat choroid plexus. Eur J Neurosci 8:629–635
Gilbert SF, Migeon BR (1975) D-valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5:11–17
Chang M, Zhang L, Tam JP, Sanders-Bush E (2000) Dissecting G protein-coupled receptor signaling pathways with membrane-permeable blocking peptides. Endogenous 5-HT(2C) receptors in choroid plexus epithelial cells. J Biol Chem 275:7021–7029
Esterle TM, Sanders-Bush E (1992) Serotonin agonists increase transferrin levels via activation of 5-HT1C receptors in choroid plexus epithelium. J Neurosci 12:4775–4782
McGrew L, Chang MS, Sanders-Bush E (2002) Phospholipase D activation by endogenous 5-hydroxytryptamine 2C receptors is mediated by Galpha13 and pertussis toxin-insensitive Gbetagamma subunits. Mol Pharmacol 62:1339–1343
Kao WW, Prockop DJ, Berg RA (1979) Kinetics for the secretion of nonhelical procollagen by freshly isolated tendon cells. J Biol Chem 254:2234–2243
Peiser C, McGregor GP, Lang RE (2000) Binding and internalization of leptin by porcine choroid plexus cells in culture. Neurosci Lett 283:209–212
Spector R (1982) Pharmacokinetics and metabolism of cytosine arabinoside in the central nervous system. J Pharmacol Exp Ther 222:1–6
Gabrion JB, Herbute S, Bouille C, Maurel D, Kuchler-Bopp S, Laabich A, Delaunoy JP (1998) Ependymal and choroidal cells in culture: characterization and functional differentiation. Microsc Res Tech 41:124–157
Hartter S, Huwel S, Lohmann T, Abou El Ela A, Langguth P, Hiemke C, Galla HJ (2003) How does the benzamide antipsychotic amisulpride get into the brain? – An in vitro approach comparing amisulpride with clozapine. Neuropsychopharmacology 28:1916–1922
Ramanathan VK, Chung SJ, Giacomini KM, Brett CM (1997) Taurine transport in cultured choroid plexus. Pharm Res 14:406–409
Gee P, Rhodes CH, Fricker LD, Angeletti RH (1993) Expression of neuropeptide processing enzymes and neurosecretory proteins in ependyma and choroid plexus epithelium. Brain Res 617:238–248
Albert O, Ancellin N, Preisser L, Morel A, Corman B (1999) Serotonin, bradykinin and endothelin signalling in a sheep choroid plexus cell line. Life Sci 64:859–867
Angelova K, Fralish GB, Puett D, Narayan P (1996) Identification of conventional and novel endothelin receptors in sheep choroid plexus cells. Mol Cell Biochem 159:65–72
Dickinson KE, Baska RA, Cohen RB, Bryson CC, Smith MA, Schroeder K, Lodge NJ (1998) Identification of [3H]P1075 binding sites and P1075-activated K+ currents in ovine choroid plexus cells. Eur J Pharmacol 345:97–101
Thornwall M, Chhajlani V, Le Greves P, Nyberg F (1995) Detection of growth hormone receptor mRNA in an ovine choroid plexus epithelium cell line. Biochem Biophys Res Commun 217:349–353
Ishiwata I, Ishiwata C, Ishiwata E, Sato Y, Kiguchi K, Tachibana T, Hashimoto H, Ishikawa H (2005) Establishment and characterization of a human malignant choroids plexus papilloma cell line (HIBCPP). Hum Cell 18:67–72
Nakashima N, Goto K, Tsukidate K, Sobue M, Toida M, Takeuchi J (1983) Choroid plexus papilloma. Light and electron microscopic study. Virchows Arch A Pathol Anat Histopathol 400:201–211
Takahashi K, Satoh F, Hara E, Murakami O, Kumabe T, Tominaga T, Kayama T, Yoshimoto T, Shibahara S (1997) Production and secretion of adrenomedullin by cultured choroid plexus carcinoma cells. J Neurochem 68:726–731
Szmydynger-Chodobska J, Pascale CL, Pfeffer AN, Coulter C, Chodobski A (2007) Expression of junctional proteins in choroid plexus epithelial cell lines: a comparative study. Cerebrospinal Fluid Res 4:11
Enjoji M, Iwaki T, Hara H, Sakai H, Nawata H, Watanabe T (1996) Establishment and characterization of choroid plexus carcinoma cell lines: connection between choroid plexus and immune systems. Jpn J Cancer Res 87:893–899
Enjoji M, Iwaki T, Nawata H, Watanabe T (1995) IgH intronic enhancer element HE2 (mu B) functions as a cis-activator in choroid plexus cells at the cellular level as well as in transgenic mice. J Neurochem 64:961–966
Schell TD, Mylin LM, Georgoff I, Teresky AK, Levine AJ, Tevethia SS (1999) Cytotoxic T-lymphocyte epitope immunodominance in the control of choroid plexus tumors in simian virus 40 large T antigen transgenic mice. J Virol 73:5981–5993
Kitazawa T, Hosoya K, Watanabe M, Takashima T, Ohtsuki S, Takanaga H, Ueda M, Yanai N, Obinata M, Terasaki T (2001) Characterization of the amino acid transport of new immortalized choroid plexus epithelial cell lines: a novel in vitro system for investigating transport functions at the blood–cerebrospinal fluid barrier. Pharm Res 18:16–22
Zheng W, Zhao Q (2002) Establishment and characterization of an immortalized Z310 choroidal epithelial cell line from murine choroid plexus. Brain Res 958:371–380
Battle T, Preisser L, Marteau V, Meduri G, Lambert M, Nitschke R, Brown PD, Corman B (2000) Vasopressin V1a receptor signaling in a rat choroid plexus cell line. Biochem Biophys Res Commun 275:322–327
Ohtsuki S, Takizawa T, Takanaga H, Terasaki N, Kitazawa T, Sasaki M, Abe T, Hosoya K, Terasaki T (2003) In vitro study of the functional expression of organic anion transporting polypeptide 3 at rat choroid plexus epithelial cells and its involvement in the cerebrospinal fluid-to-blood transport of estrone-3-sulfate. Mol Pharmacol 63:532–537
Terasaki T, Ohtsuki S, Hori S, Takanaga H, Nakashima E, Hosoya K (2003) New approaches to in vitro models of blood–brain barrier drug transport. Drug Discov Today 8:944–954
Miettinen M, Clark R, Virtanen I (1986) Intermediate filament proteins in choroid plexus and ependyma and their tumors. Am J Pathol 123:231–240
Peraldi-Roux S, Nguyen-Than Dao B, Hirn M, Gabrion J (1990) Choroidal ependymocytes in culture: expression of markers of polarity and function. Int J Dev Neurosci 8:575–588
Nataf S, Strazielle N, Hatterer E, Mouchiroud G, Belin MF, Ghersi-Egea JF (2006) Rat choroid plexuses contain myeloid progenitors capable of differentiation toward macrophage or dendritic cell phenotypes. Glia 54:160–171
Sousa JC, Cardoso I, Marques F, Saraiva MJ, Palha JA (2007) Transthyretin and Alzheimer’s disease: where in the brain? Neurobiol Aging 28:713–718
Richardson SJ (2009) Evolutionary changes to transthyretin: evolution of transthyretin biosynthesis. FEBS J 276:5342–5356
Thomas T, Stadler E, Dziadek M (1992) Effects of the extracellular matrix on fetal choroid plexus epithelial cells: changes in morphology and multicellular organization do not affect gene expression. Exp Cell Res 203:198–213
Ghersi-Egea JF, Strazielle N, Murat A, Jouvet A, Buenerd A, Belin MF (2006) Brain protection at the blood–cerebrospinal fluid interface involves a glutathione-dependent metabolic barrier mechanism. J Cereb Blood Flow Metab 26:1165–1175
Gazzin S, Strazielle N, Schmitt C, Fevre-Montange M, Ostrow JD, Tiribelli C, Ghersi-Egea JF (2008) Differential expression of the multidrug resistance-related proteins ABCb1 and ABCc1 between blood–brain interfaces. J Comp Neurol 510:497–507
Tu GF, Achen MG, Aldred AR, Southwell BR, Schreiber G (1991) The distribution of cerebral expression of the transferrin gene is species specific. J Biol Chem 266:6201–6208
Lippoldt A, Jansson A, Kniesel U, Andbjer B, Andersson A, Wolburg H, Fuxe K, Haller H (2000) Phorbol ester induced changes in tight and adherens junctions in the choroid plexus epithelium and in the ependyma. Brain Res 854:197–206
Lippoldt A, Liebner S, Andbjer B, Kalbacher H, Wolburg H, Haller H, Fuxe K (2000) Organization of choroid plexus epithelial and endothelial cell tight junctions and regulation of claudin-1, -2 and -5 expression by protein kinase C. Neuroreport 11:1427–1431
Furuse M, Sasaki H, Tsukita S (1999) Manner of interaction of heterogeneous claudin species within and between tight junction strands. J Cell Biol 147:891–903
Tsukita S, Furuse M (2002) Claudin-based barrier in simple and stratified cellular sheets. Curr Opin Cell Biol 14:531–536
Strazielle N, Belin MF, Ghersi-Egea JF (2003) Choroid plexus controls brain availability of anti-HIV nucleoside analogs via pharmacologically inhibitable organic anion transporters. AIDS 17:1473–1485
Saito Y, Wright EM (1983) Bicarbonate transport across the frog choroid plexus and its control by cyclic nucleotides. J Physiol 336:635–648
Welch K, Araki H (1975) Features of the choroid plexus of the cat studied in vitro. In: Cserr HF, Fenstermacher JD, Fencl V (eds) Fluid environment of the brain. Academic, New York, pp 157–165
Wright EM (1972) Mechanisms of ion transport across the choroid plexus. J Physiol 226:545–571
Claude P, Goodenough DA (1973) Fracture faces of zonulae occludentes from “tight” and “leaky” epithelia. J Cell Biol 58:390–400
Welch K, Sadler K (1966) Permeability of the choroid plexus of the rabbit to several solutes. Am J Physiol 210:652–660
Lo CM, Keese CR, Giaever I (1999) Cell-substrate contact: another factor may influence transepithelial electrical resistance of cell layers cultured on permeable filters. Exp Cell Res 250:576–580
Papenheimer JR (1961) Active transport of Diodrast and phenolsulfonphtalein from cerebrospinal fluid to blood. Am J Physiol 200:1–10
Ennis SR, Novotny A, Xiang J, Shakui P, Masada T, Stummer W, Smith DE, Keep RF (2003) Transport of 5-aminolevulinic acid between blood and brain. Brain Res 959:226–234
Redzic ZB, Isakovic AJ, Misirlic Dencic ST, Popadic D, Segal MB (2006) Uneven distribution of nucleoside transporters and intracellular enzymatic degradation prevent transport of intact [14C] adenosine across the sheep choroid plexus epithelium as a monolayer in primary culture. Cerebrospinal Fluid Res 3:4
Isakovic AJ, Dencic SM, Segal MB, Redzic ZB (2008) Transport of [14C]hypoxanthine by sheep choroid plexus epithelium as a monolayer in primary culture: Na+-dependent and Na+-independent uptake by the apical membrane and rapid intracellular metabolic conversion to nucleotides. Neurosci Lett 431:135–140
Hakvoort A, Haselbach M, Galla HJ (1998) Active transport properties of porcine choroid plexus cells in culture. Brain Res 795:247–256
Zheng W, Blaner WS, Zhao Q (1999) Inhibition by lead of production and secretion of transthyretin in the choroid plexus: its relation to thyroxine transport at blood–CSF barrier. Toxicol Appl Pharmacol 155:24–31
Steffen BJ, Breier G, Butcher EC, Schulz M, Engelhardt B (1996) ICAM-1, VCAM-1, and MAdCAM-1 are expressed on choroid plexus epithelium but not endothelium and mediate binding of lymphocytes in vitro. Am J Pathol 148:1819–1838
Pappenheimer JR, Renkin EM, Borrero LM (1951) Filtration, diffusion and molecular sieving through peripheral capillary membranes; a contribution to the pore theory of capillary permeability. Am J Physiol 167:13–46
Khuth ST, Strazielle N, Giraudon P, Belin MF, Ghersi-Egea JF (2005) Impairment of blood–cerebrospinal fluid barrier properties by retrovirus-activated T lymphocytes: reduction in cerebrospinal fluid-to-blood efflux of prostaglandin E2. J Neurochem 94:1580–1593
Acknowledgements
This work was supported by the European Union (HEALTH-F2-2009-241778).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Strazielle, N., Ghersi-Egea, JF. (2011). In Vitro Models of the Blood–Cerebrospinal Fluid Barrier and Their Use in Neurotoxicological Research. In: Aschner, M., Suñol, C., Bal-Price, A. (eds) Cell Culture Techniques. Neuromethods, vol 56. Humana Press. https://doi.org/10.1007/978-1-61779-077-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-077-5_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-076-8
Online ISBN: 978-1-61779-077-5
eBook Packages: Springer Protocols