细胞生物学
生物
脉络丛
信号转导
细胞内
ATP结合盒运输机
运输机
生物化学
神经科学
基因
中枢神经系统
作者
Louise Hulme,Alexandra Hochstetler,Christian Schwerk,Horst Schroten,Hiroshi Ishikawa,Chun‐Yu Tung,Benjamin J. Perrin,Bonnie L. Blazer‐Yost
出处
期刊:American Journal of Physiology-cell Physiology
[American Physiological Society]
日期:2022-12-01
卷期号:323 (6): C1823-C1842
被引量:5
标识
DOI:10.1152/ajpcell.00193.2022
摘要
The objectives of these studies were twofold: 1) to characterize the human choroid plexus papilloma (HIBCPP) cell line as a model of the blood-cerebrospinal fluid barrier (BCSFB) via morphology, tightness, and polarization of transporters in choroid plexus epithelia (CPe), and 2) to utilize Ussing-style electrophysiology to elucidate signaling pathways associated with the activation of the transient receptor potential vanilloid 4 (TRPV4) channel involved in cerebrospinal fluid (CSF) secretion. RT-PCR was implemented to determine gene expression of cell fate markers, junctional complex proteins, and transporters of interest. Scanning electron microscopy and confocal three-dimensional renderings of cultures grown on permeable supports were utilized to delineate the morphology of the brush border, junctional complexes, and polarization of key transporters. Electrophysiology was used to understand and explore TRPV4-mediated signaling in the HIBCPP cell line, considering both short-circuit current ( I sc ) and conductance responses. HIBCPP cells grown under optimized culture conditions exhibited minimal multilayering, developed an intermediate resistance monolayer, retained differentiation properties, and expressed, and correctly localized, junctional proteins and native transporters. We found that activation of TRPV4 resulted in a robust, multiphasic change in electrogenic ion flux and increase in conductance accompanied by substantial fluid secretion. This response appears to be modulated by a number of different effectors, implicating phospholipase C (PLC), protein kinase C (PKC), and phosphoinositide 3-kinase (PI3K) in TRPV4-mediated ion flux. The HIBCPP cell line is a representative model of the human BCSFB, which can be utilized for studies of transporter function, intracellular signaling, and regulation of CSF production.
科研通智能强力驱动
Strongly Powered by AbleSci AI