title: Channel-mediated water movement across enclosed or perfused mouse intrahepatic bile duct units.
abstract: We previously reported the development of reproducible techniques for isolating and perfusing intact intrahepatic bile duct units (IBDUs) from rats. Given the advantages of transgenic and knockout mice for exploring ductal bile formation, we report here the adaptation of those techniques to mice and their initial application to the study of water transport across mouse intrahepatic biliary epithelia. IBDUs were isolated from livers of normal mice by microdissection combined with enzymatic digestion. After culture, isolated IBDUs sealed to form intact, polarized compartments, and a microperfusion system employing those isolated IBDUs developed. A quantitative image analysis technique was used to observe a rapid increase of luminal area when sealed IBDUs were exposed to a series of inward osmotic gradients reflecting net water secretion; the choleretic agonists secretin and forskolin also induced water secretion into IBDUs. The increase of IBDU luminal area induced by inward osmotic gradients and choleretic agonists was reversibly inhibited by HgCl2, a water channel inhibitor. With the use of a quantitative epifluorescence technique in perfused mouse IBDUs, a high osmotic water permeability (P(f) = 2.5-5.6 x 10(-2) cm/s) was found in response to osmotic gradients, further supporting the presence of water channels. These findings suggest that, as in the rat, water transport across intrahepatic biliary epithelia in mice is water channel mediated.
authors.affiliation: Center for Basic Research in Digestive Diseases, Division of Gastroenterology and Hepatology, Mayo Medical School, Clinic, and Foundation, Rochester, Minnesota 55905, USA.
authors.forename: Ai-Yu
authors.initials: AY
authors.lastname: Gong
authors.forename: Anatoly I
authors.initials: AI
authors.lastname: Masyuk
authors.forename: Patrick L
authors.initials: PL
authors.lastname: Splinter
authors.forename: Robert C
authors.initials: RC
authors.lastname: Huebert
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Tietz
authors.forename: Nicholas F
authors.initials: NF
authors.lastname: LaRusso
mesh_terms: D000818:Animals; D020346:Aquaporins / Q000378:metabolism*; D001653:Bile Ducts, Intrahepatic / Q000187:drug effects / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002756:Cholagogues and Choleretics / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D046508:Culture Techniques; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D008627:Mercuric Chloride / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D009995:Osmosis; D010477:Perfusion; D012633:Secretin / Q000494:pharmacology; D014867:Water / Q000378:metabolism*
chemical_list: D020346:Aquaporins; D002756:Cholagogues and Choleretics; D014867:Water; D012633:Secretin; D005576:Colforsin; D008627:Mercuric Chloride