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Prostaglandins Stimulate cAMP Generation via Serosal Membrane Receptors in Porcine Epithelial Cells
Author(s) -
Riley Melissa,
Schultz Bruce
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.1007.10
Subject(s) - adenylyl cyclase , cystic fibrosis transmembrane conductance regulator , receptor , endocrinology , medicine , vas deferens , cyclic adenosine monophosphate , prostaglandin , agonist , second messenger system , camp dependent pathway , protein kinase a , prostaglandin e2 , adenosine , chemistry , prostaglandin e2 receptor , purinergic receptor , biology , microbiology and biotechnology , kinase , cystic fibrosis
Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a gene that codes for an anion channel that is regulated by cyclic adenosine monophosphate (cAMP) and its associated kinase, protein kinase A (PKA). Numerous physiological agonists are known to stimulate the activity of adenylyl cyclase, which generates cAMP. Our previous work demonstrated that acute exposure to prostaglandin (PG) D 2 or PGE 2 causes a rapid and sustained increase in short circuit current ( I sc ), an indicator of anion secretion, across cultured pig vas deferens epithelial cells, an outcome that is consistent with PG‐induced increases in adenylyl cyclase activity. Also, it was shown that indomethacin, an agent that blocks prostaglandin synthesis, precluded the effects of the stimulant bradykinin, which suggests that all components of the prostaglandin synthesis and the response pathways are present in these cells. However, the molecular identity of the prostaglandin receptor(s), the location(s) of these receptors and second messengers that mediate the responses remain to be determined. Thus, experiments were carried out to determine to localize receptors of PGD 2 and PGE 2 to either the mucosal or serosal aspect of the cells. These experiments were carried out with cultured primary adult pig vas deferens epithelial (1°PVD) cells using a modified Ussing‐style flux chamber. Serosal, but not mucosal exposure of PGE 2 produced a rapid increase in I sc , suggesting serosal localization of EP receptors. Similarly, PGD 2 and a selective DP receptor agonist, BW 245C, produced an effect following serosal exposure. Experiments were also carried out to determine the effect of PGs on cAMP generation using a cAMP enzyme immunoassay kit with 1°PVD cells. PGE2, PGD2, and BW 245C all showed an increase in stimulation of cAMP generation. This stimulation was inhibited by the presence of DP receptor antagonist, BW A868C, and EP2/EP4 receptor antagonists, PF‐04418948 and L‐161,982. The simplest interpretation of data is that both EP and DP receptors are localized to the serosal membrane of vas deferens epithelial cells and cAMP generation is affected by inhibition of these receptors. Activation of these receptors is tightly linked to anion secretion, which would be expected to increase pH and fluid volume in the male reproductive duct. Support or Funding Information IOS‐1238831