title: Effect of acetazolamide on aquaporin-1 and fluid flow in cultured choroid plexus.
abstract: Acetazolamide (AZA), used in treatment of early or infantile hydrocephalus, is effective in some cases, while its effect on the choroid plexus (CP) remains ill-defined. The drug reversibly inhibits aquaporin-4 (AQP4), the most ubiquitous "water pore" in the brain, and perhaps modulation of AQP1 (located apically on CP cells) by AZA may reduce cerebrospinal fluid (CSF) production. We sought to elucidate the effect of AZA on AQP1 and fluid flow in CP cell cultures.CP tissue culture from 10-day Sprague-Dawley rats and a TRCSF-B cell line were grown on Transwell permeable supports and treated with 100 μM AZA. Fluid assays to assess direction and extent of fluid flow, and AQP1 expression patterns by immunoblot, Immuncytochemistry (ICC), and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) were performed.Immunoblots and ICC analyses showed a decrease in AQP1 protein shortly after AZA treatment (lowest at 12 h), with transient AQP1 reduction mediated by mRNA expression (lowest at 6 h). Transwell fluid assays indicated a fluid shift at 2 h, before significant changes in AQP1 mRNA or protein levels.Timing of AZA effect on AQP1 suggests the drug alters protein transcription, while affecting fluid flow by a concomitant method. It is plausible that other mechanisms account for these phenomena, as the processes may occur independently.
authors: Burnett School of Biomedical Sciences, University of Central Florida, Orlando, FL, USA.
authors: Pouya A
authors: PA
authors: Ameli
authors: Meenu
authors: M
authors: Madan
authors: Srinivasulu
authors: S
authors: Chigurupati
authors: Amin
authors: A
authors: Yu
authors: Sic L
authors: SL
authors: Chan
authors: Jogi V
authors: JV
authors: Pattisapu
mesh_terms: D000086:Acetazolamide / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D051398:Aquaporin 1 / Q000378:metabolism*; D002199:Capillary Permeability / Q000187:drug effects; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology*; D002831:Choroid Plexus / Q000187:drug effects* / Q000378:metabolism; D003911:Dextrans; D005786:Gene Expression Regulation / Q000187:drug effects*; D057446:Hydrodynamics; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012235:Rhodamines; D013997:Time Factors
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D003911:Dextrans; D012333:RNA, Messenger; D012235:Rhodamines; C539719:dextran tetramethylrhodamine; D051398:Aquaporin 1; D000086:Acetazolamide