Functional characterization and localization of AQP3 in the human colon.
Water channels or aquaporins (AQPs) have been identified in a large variety of tissues. Nevertheless, their role in the human gastrointestinal tract, where their action is essential for the reabsorption and secretion of water and electrolytes, is still unclear. The purpose of the present study was to investigate the structure and function of water channels expressed in the human colon. A cDNA fragment of about 420 bp with a 98% identity to human AQP3 was amplified from human stomach, small intestine and colon by reverse transcription polymerase chain reaction (RT-PCR) and a transcript of 2.2 kb was expressed more abundantly in colon than in jejunum, ileum and stomach as indicated by Northern blots. Expression of mRNA from the colon of adults and children but not from other gastrointestinal regions in Xenopus oocytes enhanced the osmotic water permeability, and the urea and glycerol transport in a manner sensitive to an antisense AQP3 oligonucleotide, indicating the presence of functional AQP3. Immunocytochemistry and immunofluorescence studies in human colon revealed that the AQP3 protein is restricted to the villus epithelial cells. The immunostaining within these cells was more intense in the apical than in the basolateral membranes. The presence of AQP3 in villus epithelial cells suggests that AQP3 is implicated in water absorption across human colonic surface cells.
Silberstein
C
C
Laboratorio de Fisiopatogenia, Departamento de Fisiología, Facultad de Medicina, Universidad Nacional de Buenos Aires, Buenos Aires, Argentina.
Kierbel
A
A
Amodeo
G
G
Zotta
E
E
Bigi
F
F
Berkowski
D
D
Ibarra
C
C
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D051400:Aquaporin 3; D020346:Aquaporins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D015152:Blotting, Northern; D002463:Cell Membrane Permeability; D002648:Child; D002675:Child, Preschool; D003106:Colon / Q000737:chemistry*; D004847:Epithelial Cells / Q000737:chemistry; D015200:Fluoroimmunoassay; D006801:Humans; D007150:Immunohistochemistry; D007408:Intestinal Absorption / Q000502:physiology*; D008969:Molecular Sequence Data; D009865:Oocytes; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014982:Xenopus laevis
C496027:AQP3 protein, human; D020346:Aquaporins; D012333:RNA, Messenger; D051400:Aquaporin 3