Identification of transport abnormalities in duodenal mucosa and duodenal enterocytes from patients with cystic fibrosis.
OBJECTIVE
The duodenum is a cystic fibrosis transmembrane conductance regulator (CFTR)-expressing epithelium with high bicarbonate secretory capacity. We aimed to define the role of CFTR in human duodenal epithelial bicarbonate secretion in normal (NL) subjects and patients with cystic fibrosis (CF).


METHODS
Endoscopic biopsy specimens of the duodenal bulb were obtained from 9 CF patients and 16 volunteers. Tissues were mounted in modified Ussing chambers. Bicarbonate secretion and short-circuit current (Isc) were quantitated under basal conditions and in response to dibutyryl adenosine 3',5'-cyclic monophosphate (db-cAMP), carbachol, and the heat-stable toxin of Escherichia coli (STa). Duodenocytes were also isolated and loaded with the pH-sensitive fluoroprobe BCECF/AM, and intracellular pH (pH(i)) was measured at rest and after intracellular acidification and alkalinization.


RESULTS
Basal HCO(3)(-) secretion and Isc were significantly lower in the CF vs. NL duodenal mucosa. In contrast to NL, db-cAMP failed to alter either HCO(3)(-) or Isc in CF tissues. However, in CF, carbachol resulted in an electroneutral HCO(3)(-) secretion, whereas STa induced electrogenic HCO(3)(-) secretion that was similar to NL. In CF and NL duodenocytes, basal pH(i) and recovery from an acid load were comparable, but pH(i) recovery after an alkaline load in CF duodenocytes was Cl(-) dependent, whereas in NL duodenocytes it was Cl(-) independent.


CONCLUSIONS
These findings implicate CFTR in NL duodenal alkaline transport and its absence in CF. Although duodenal bicarbonate secretion is impaired in CF tissues, alternate pathway(s) likely exist that can be activated by carbachol and STa.
Pratha
V S
VS
Division of Gastroenterology, Department of Medicine, University of California at San Diego, San Diego, California, USA.
Hogan
D L
DL
Martensson
B A
BA
Bernard
J
J
Zhou
R
R
Isenberg
J I
JI
D000136:Acid-Base Equilibrium / Q000502:physiology; D000143:Acids / Q000378:metabolism; D000328:Adult; D000468:Alkalies / Q000378:metabolism; D000643:Ammonium Chloride / Q000494:pharmacology; D017920:Antiporters / Q000378:metabolism; D001427:Bacterial Toxins / Q000494:pharmacology; D001639:Bicarbonates / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D001706:Biopsy; D003994:Bucladesine / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D027963:Chloride-Bicarbonate Antiporters; D002712:Chlorides / Q000378:metabolism; D018679:Cholinergic Agonists / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004386:Duodenum / Q000378:metabolism* / Q000473:pathology; D020895:Enterocytes / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D004768:Enterotoxins / Q000494:pharmacology; D029968:Escherichia coli Proteins; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007408:Intestinal Absorption / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D011422:Propionates / Q000494:pharmacology
D000143:Acids; D000468:Alkalies; D017920:Antiporters; D001427:Bacterial Toxins; D001639:Bicarbonates; C505032:CFTR protein, human; D027963:Chloride-Bicarbonate Antiporters; D002712:Chlorides; D018679:Cholinergic Agonists; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011422:Propionates; C034151:heat stable toxin (E coli); D000643:Ammonium Chloride; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D003994:Bucladesine; D002217:Carbachol; C029658:propionic acid