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cotransporter activity in isolated, polarized choroid plexus
cells
Anesthesiology Research Division, Departments of 1 Anesthesiology and 3 Pharmacology and 2 Division of Nephrology, Vanderbilt University Medical Center, Nashville, Tennessee 37232
The function of the apical
Na+-K+-2Cl
cotransporter in mammalian choroid plexus (CP) is uncertain and
controversial. To investigate cotransporter function, we developed a
novel dissociated rat CP cell preparation in which single, isolated
cells maintain normal polarized morphology. Immunofluorescence
demonstrated that in isolated cells the
Na+-K+-ATPase,
Na+-K+-2Cl
cotransporter, and aquaporin 1 water channel remained localized to the
brush border, whereas the
Cl/HCO3
(anion) exchanger type 2 was confined to the basolateral membrane. We
utilized video-enhanced microscopy and cell volume measurement
techniques to investigate cotransporter function. Application of 100 µM bumetanide caused CP cells to shrink rapidly. Elevation of
extracellular K+ from 3 to 6 or 25 mM caused CP cells to swell 18 and 33%, respectively. Swelling was
blocked completely by Na+ removal
or by addition of 100 µM bumetanide. Exposure of CP cells to 5 mM
BaCl2 induced rapid swelling that
was inhibited by 100 µM bumetanide. We conclude that the CP
cotransporter is constitutively active and propose that it functions in
series with Ba2+-sensitive
K+ channels to reabsorb
K+ from cerebrospinal fluid to blood.
potassium transport; cerebrospinal fluid secretion; cerebral edema; intracranial hypertension; bumetanide
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