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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

A SCL4A10 gene product maps selectively to the basolateral plasma membrane of choroid plexus epithelial cells

Water and Salt Research Center, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark

Submitted 9 June 2003 ; accepted in final form 28 October 2003

The choroid plexus epithelium of the brain ventricular system produces the majority of the cerebrospinal fluid and thereby defines the ionic composition of the interstitial fluid in the brain. The transepithelial movement of Na⁺ and water in the choroid plexus depend on a yet-unidentified basolateral stilbene-sensitive - uptake protein. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed the expression in the choroid plexus of SLC4A10 mRNA, which encodes a stilbene-sensitive - transporter. Anti-COOH-terminal antibodies were developed to determine the specific expression and localization of this - transport protein. Immunoblotting demonstrated antibody binding to a 180-kDa protein band from mouse and rat brain preparations enriched with choroid plexus. The immunoreactive band migrated as a 140-kDa protein after N-deglycosylation, consistent with the predicted molecular size of the SLC4A10 gene product. Bright-field immunohistochemistry and immunoelectron microscopy demonstrated strong labeling confined to the basolateral plasma membrane domain of the choroid plexus epithelium. Furthermore, the stilbene-insensitive - cotransporter, NBCn1, was also localized to the basolateral plasma membrane domain of the choroid plexus epithelium. Hence, we propose that the SLC4A10 gene product and NBCn1 both function as basolateral entry pathways and that the SLC4A10 gene product may be responsible for the stilbene-sensitive - uptake that is essential for cerebrospinal fluid production.

cerebrospinal fluid; bicarbonate metabolism; immunohistochemistry; electron microscopy; immunoblotting

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