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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Role of Constitutive Cyclooxygenase-2 in Prostaglandin-Dependent Secretion in Mouse Colon In Vitro

Wallace K. MacNaughton and Kelly Cushing
Journal of Pharmacology and Experimental Therapeutics May 2000, 293 (2) 539-544;
Wallace K. MacNaughton
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Kelly Cushing
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Abstract

The relative contributions of cyclooxygenase (COX)-1 and COX-2 in mediating prostaglandin (PG)-dependent chloride secretion were investigated in segments of mouse colon mounted in Ussing-type diffusion chambers. COX-2 mRNA and protein were constitutively expressed as shown by reverse transcription-polymerase chain reaction and Western immunoblot, respectively. COX-2 immunoreactivity was detected immunohistochemically in cells lying subjacent to the crypt epithelial cells. In segments of colon mounted in Ussing chambers, arachidonic acid caused a concentration-dependent increase in short-circuit current that was blocked by piroxicam, the COX-2 inhibitor NS-398, and the COX-1 inhibitor SC-560. Exposure to the PG-dependent secretagogue, bradykinin, also caused an increase in short-circuit current that was not blocked by piroxicam or SC-560, and only by the highest dose of NS-398. When incubated in the presence of 10 μM arachidonic acid, segments of mouse colon produced both PGE2 and PGD2. Synthesis of PGE2but not PGD2 was blocked by NS-398 and SC-560. These data demonstrate that both COX-1 and COX-2 are constitutively expressed in the mouse colon, and both contribute to PG-dependent electrolyte transport.

Footnotes

  • Send reprint requests to: Wallace MacNaughton, Ph.D., Department of Physiology and Biophysics, University of Calgary, 3330 Hospital Dr. NW, Calgary, Alberta, Canada T2N 4N1. E-mail:wmacnaug{at}ucalgary.ca

  • ↵1 This study was supported by a New Investigator Establishment grant from the Canadian Association of Gastroenterology and the Medical Research Council of Canada.

  • ↵2 W.K.M. is an Alberta Heritage Foundation for Medical Research Scholar.

  • Abbreviations:
    PG
    prostaglandin
    COX
    cyclooxygenase
    RT-PCR
    reverse transcription-polymerase chain reaction
    Isc
    short-circuit current
    IR
    immunoreactivity
    • Received August 2, 1999.
    • Accepted January 12, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 293 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 293, Issue 2
1 May 2000
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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Role of Constitutive Cyclooxygenase-2 in Prostaglandin-Dependent Secretion in Mouse Colon In Vitro

Wallace K. MacNaughton and Kelly Cushing
Journal of Pharmacology and Experimental Therapeutics May 1, 2000, 293 (2) 539-544;

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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Role of Constitutive Cyclooxygenase-2 in Prostaglandin-Dependent Secretion in Mouse Colon In Vitro

Wallace K. MacNaughton and Kelly Cushing
Journal of Pharmacology and Experimental Therapeutics May 1, 2000, 293 (2) 539-544;
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