Culture of Choroid Plexus Epithelial Cells and In Vitro Model of Blood–CSF Barrier

Monnot, Andrew D.; Zheng, Wei

doi:10.1007/978-1-62703-125-7_2

Cited by 45 publications

(46 citation statements)

References 24 publications

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“…Recent advances in the generation of an immortalized choroid plexus cell line for use in in vitro experiments may prove extremely valuable in efforts to understand Na,K-ATPase trafficking in this tissue. 127 In the case of RPE cells, the absence of epithelial-specific m1B AP expression is associated with the aberrant apical localization of the coxsackie and adenovirus receptor protein. 128 Interestingly, knockdown experiments in MDCK cells demonstrated that m1B is required for correct recycling of coxsackie and adenovirus receptor to the basolateral membrane, but not for its initial biosynthetic delivery.…”

Section: Cell-specific Variations In Traffickingmentioning

confidence: 41%

Trafficking to the Apical and Basolateral Membranes in Polarized Epithelial Cells

Stoops

Caplan

2014

Journal of the American Society of Nephrology

100

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Renal epithelial cells must maintain distinct protein compositions in their apical and basolateral membranes in order to perform their transport functions. The creation of these polarized protein distributions depends on sorting signals that designate the trafficking route and site of ultimate functional residence for each protein. Segregation of newly synthesized apical and basolateral proteins into distinct carrier vesicles can occur at the trans-Golgi network, recycling endosomes, or a growing assortment of stations along the cellular trafficking pathway. The nature of the specific sorting signal and the mechanism through which it is interpreted can influence the route a protein takes through the cell. Cell type-specific variations in the targeting motifs of a protein, as are evident for Na,K-ATPase, demonstrate a remarkable capacity to adapt sorting pathways to different developmental states or physiologic requirements. This review summarizes our current understanding of apical and basolateral trafficking routes in polarized epithelial cells.

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Section: Cell-specific Variations In Traffickingmentioning

confidence: 41%

Trafficking to the Apical and Basolateral Membranes in Polarized Epithelial Cells

Stoops

Caplan

2014

Journal of the American Society of Nephrology

100

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“…Primary cultures of mouse astrocytes and choroid plexus epithelial cells were performed, as reported [40, 48]. In brief, the choroid plexus from the lateral ventricles and 4 th ventricle were removed from 15 C57BL6 male mice (4–8 weeks old), digested in 0.4% pronase in Hank’s balanced salt solution (HBSS) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing fetal bovine serum (FBS), epidermal growth factor (EGF) and antibiotics (penicillin/streptomycin) on collagen coated wells.…”

Section: Methodsmentioning

confidence: 99%

Glymphatic distribution of CSF-derived apoE into brain is isoform specific and suppressed during sleep deprivation

Achariyar

Peng

et al. 2016

Mol Neurodegeneration

202

137

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BackgroundApolipoprotein E (apoE) is a major carrier of cholesterol and essential for synaptic plasticity. In brain, it’s expressed by many cells but highly expressed by the choroid plexus and the predominant apolipoprotein in cerebrospinal fluid (CSF). The role of apoE in the CSF is unclear. Recently, the glymphatic system was described as a clearance system whereby CSF and ISF (interstitial fluid) is exchanged via the peri-arterial space and convective flow of ISF clearance is mediated by aquaporin 4 (AQP4), a water channel. We reasoned that this system also serves to distribute essential molecules in CSF into brain. The aim was to establish whether apoE in CSF, secreted by the choroid plexus, is distributed into brain, and whether this distribution pattern was altered by sleep deprivation.MethodsWe used fluorescently labeled lipidated apoE isoforms, lenti-apoE3 delivered to the choroid plexus, immunohistochemistry to map apoE brain distribution, immunolabeled cells and proteins in brain, Western blot analysis and ELISA to determine apoE levels and radiolabeled molecules to quantify CSF inflow into brain and brain clearance in mice. Data were statistically analyzed using ANOVA or Student’s t- test.ResultsWe show that the glymphatic fluid transporting system contributes to the delivery of choroid plexus/CSF-derived human apoE to neurons. CSF-delivered human apoE entered brain via the perivascular space of penetrating arteries and flows radially around arteries, but not veins, in an isoform specific manner (apoE2 > apoE3 > apoE4). Flow of apoE around arteries was facilitated by AQP4, a characteristic feature of the glymphatic system. ApoE3, delivered by lentivirus to the choroid plexus and ependymal layer but not to the parenchymal cells, was present in the CSF, penetrating arteries and neurons. The inflow of CSF, which contains apoE, into brain and its clearance from the interstitium were severely suppressed by sleep deprivation compared to the sleep state.ConclusionsThus, choroid plexus/CSF provides an additional source of apoE and the glymphatic fluid transporting system delivers it to brain via the periarterial space. By implication, failure in this essential physiological role of the glymphatic fluid flow and ISF clearance may also contribute to apoE isoform-specific disorders in the long term.Electronic supplementary materialThe online version of this article (doi:10.1186/s13024-016-0138-8) contains supplementary material, which is available to authorized users.

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“…Thus, Z310 cells have the same polygonal morphology of CP epithelial cells, forming confluent monolayers as the primary cell cultures. The cell line Z310 was cultured as described previously [67].…”

Section: Cell Culture and Sirna Knockdownsupporting

confidence: 48%

‘Smelling’ the cerebrospinal fluid: olfactory signaling molecules are expressed in and mediate chemosensory signaling from the choroid plexus

et al. 2016

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The olfactory-type signaling machinery has been known to be involved not only in odorant detection but also in other tissues with unsuspected sensory roles. As a barrier, the choroid plexus (CP) is an active participant in the monitoring of the cerebrospinal fluid (CSF), promptly responding to alterations in its composition. We hypothesized that olfactory signaling could be active in CP, contributing to the surveillance of the CSF composition. We determined the mRNA and protein expression of the major components of the olfactory transduction pathway in the rat CP, including odorant receptors, the olfactory G-protein (Gaolf), adenylate cyclase 3 and cyclic nucleotide-gated channel 2. The functionality of the transduction pathway and the intracellular mechanisms involved were analyzed by DC field potential recording electrophysiological analysis, in an ex vivo CP-brain setup, using polyamines as stimuli and blockers of the downstream signaling pathways. Concentration-dependent responses were obtained for the polyamines studied (cadaverine, putrescine, spermine and spermidine), all known to be present in the CSF. Transfection of a CP epithelial cell line with siRNA against Gaolf effectively knocked down protein expression and reduced the CP cells' response to spermine. Thus, the key components of the olfactory chemosensory apparatus are present and are functional in murine CP, and polyamines seem to trigger both the cAMP and the phospholipase C-inositol 1,4,5-trisphosphate pathways. Olfactory-like chemosensory signaling may be an essential component of the CP chemical surveillance apparatus to detect alterations in the CSF composition, and to elicit responses to modulate and maintain brain homeostasis.

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Culture of Choroid Plexus Epithelial Cells and In Vitro Model of Blood–CSF Barrier

Cited by 45 publications

References 24 publications

Trafficking to the Apical and Basolateral Membranes in Polarized Epithelial Cells

Trafficking to the Apical and Basolateral Membranes in Polarized Epithelial Cells

Glymphatic distribution of CSF-derived apoE into brain is isoform specific and suppressed during sleep deprivation

‘Smelling’ the cerebrospinal fluid: olfactory signaling molecules are expressed in and mediate chemosensory signaling from the choroid plexus

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