Laboratory Investigation

Kidney International (1993) 43, 1286–1297; doi:10.1038/ki.1993.181

ADH resistance of LLC-PK1 cells caused by overexpression of cAMP-phosphodiesterase type-IV

Mario Yamaki, Steven McIntyre, Josie M Murphy, Johannes V Swinnen, Marco Conti and Thomas P Dousa

Nephrology Research Unit, Division of Nephrology, Mayo Clinic and Foundation, Rochester, Minnesota, and Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina, USA

Correspondence: Thomas P Dousa MD PhD, Mayo Clinic and Foundation, 901 Guggenheim Building, 200 First Street SW, Rochester, Minnesota 55905, USA.

Received 22 July 1992; Revised 12 January 1993; Accepted 14 January 1993.

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Abstract

ADH resistance of LLC-PK1 cells caused by overexpression of cAMP-phosphodiesterase type-IV. The studies of animal models of nephrogenic diabetes insipidus (NDI) suggest that abnormally high activity of cAMP phosphodiesterase (cAMP-PDE) may cause unresponsiveness to the diuretic effect of AVP. We explored whether overexpression of one of the cAMP-PDE type isozymes, PDE-IV, in [8-Arg]-vasopressin (AVP) sensitive renal epithelial LLC-PK1 cells can prevent the hormone-elicited cAMP increase. LLC-PK1 cells were stably transfected with ratPDE3.1 cDNA (which encodes for rolipram-sensitive PDE-IV), inserted in plasmid pCMV5 and then were compared with sham-transfected LLC-PK1 cells and wild LLC-PK1 cells. In the stably transfected clone (LLC-PK1S#16), the rolipram-sensitive PDE-IV activity was about five times higher than in controls, whereas activities of other types of PDEs were not different. The presence of cognate mRNA for PDE-IV was confirmed by Northern blot. Whereas in the control cells (wild LLC-PK1 cells and sham-transfected LLC-PK1 cells), the incubation with 10-7 M AVP increased cAMP more than tenfold, the LLC-PK1S#16 cells with overexpressed cAMP-PDE were resistant to cAMP-increasing effects of AVP and forskolin. However, in the same LLC-PK1-S#16 cells the cGMP increases in response to nitroprusside were not diminished. The AVP-dependent cAMP accumulation in LLC-PK1S#16 cells with overexpressed PDE-IV was restored by addition of roliprams which decreased cAMP-PDE activity to the levels similar to those in wild LLC-PK1 cells and sham-transfected LLC-PK1-#A1 cells. In contrast, inhibitors of other PDE isozymes (PDE-I or PDE-III) had little or no effect. Our findings show that excessive activity of cAMP-PDE, in this case of isozyme PDE-IV, can cause resistance to AVP which is analogous to that observed in collecting ducts of mice with hereditary nephrogenic diabetes insipidus.

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References

  1. Beavo JA: Multiple phosphodiesterase isoenzymes; background, nomenclature and implications, in Cyclic Nucleotide Phosphodiesterases: Structure, Regulation and Drug Action, edited by Beavo J, Houslay M, London, John Wiley & Sons, 1990, pp. 3–5
  2. Beavo JA: Multiple isozymes of cyclic nucleotide phosphodiesterase. Adv Sec Mess Phosphoprotein Res 22:1–38, 1988
  3. Beavo JA, Reifsnyder DH: Primary sequence of cyclic nucleotide phosphodiesterase isozymes and the design of seletive inhibitors. Trend Pharmacol Sci 11:150–155, 1990
  4. Conti M, Jin S-LC, Monaco L, Repaske DR, Swinnen JV: Hormonal regulation of cyclic nucleotide phosphodiesterases. Endocr Rev 12:218–234, 1991 | PubMed | ChemPort |
  5. Conti M, Swinnen JV, Tsikalas KE, Jin S-LC: Structure and regulation of the rat high-affinity cyclic AMP phosphodiesterases, in Advances in Second Messenger and Phosphoprotein Research (vol. 25), edited by Strada S, Hidaka H, New York, Raven Press, Ltd., 1992, pp. 87–99
  6. Nicholson CD, Challiss RAJ, Shahid M: Differential modulation of tissue function and therapeutic potential of selective inhibitors of cyclic nucleotide phosphodiesterase isoenzymes. Trend Pharmacol Sci 12:19–27, 1991
  7. Jackson B, Braun-Werness JL, Kusano E, Dousa TP: Concentrating defect in the adrenalectomized rat. J Clin Invest 72:997–1004, 1983 | ChemPort |
  8. Homma S, Gapstur SM, Coffey A, Valtin H, Dousa TP: Role of cAMP-phosphodiesterase isozymes in pathogenesis of murine nephrogenic diabetes insipidus. Am J Physiol 261:F345–F353, 1991 | ChemPort |
  9. Coffey AK, O'Sullivan DJ, Homma S, Dousa TP, Valtin H: Induction of intramembranous particle clusters in mice with nephrogenic diabetes insipidus. Am J Physiol 261:F640–F646, 1991 | ChemPort |
  10. Takeda S, Lin C-T, Morgano PG, McIntyre SJ, Dousa TP: High activity of low-Michaelis-Menten constant 3',5'-cyclic adenosine monophosphate-phosphodiesterase isozymes in renal inner medulla of mice with hereditary nephrogenic diabetes insipidus. Endocrinology 129:287–294, 1991 | ChemPort |
  11. Jackson BA, Edwards RM, Valtin H, Dousa TP: Cellular action of vasopressin in medullary tubules of mice with hereditary nephrogenic diabetes insipidus. J Clin Invest 54:753–762, 1980
  12. Weishaar RE, Cain MH, Bristol JA: A new generation of phosphodiesterase inhibitors: Multiple molecular forms of phosphodiesterase and the potential for drug selectivity. J Med Chem 28:537–545, 1985 | Article | PubMed | ChemPort |
  13. Swinnen SV, D'Souza B, Conti M, Ascoli M: Attenuation of cAMP-mediated responses in MA-10 Leydig tumor cells by genetic manipulation of a cAMP-phosphodiesterase. J Biol Chem 266:14383–14389, 1991
  14. Swinnen JV, Joseph DR, Conti M: The mRNA encoding a high-affinity cAMP phosphodiesterase is regulated by hormones and cAMP. Proc Natl Acad Sci USA 86:8197–8201, 1989 | PubMed | ChemPort |
  15. Livi GP, Kmetz P, McHale MM, Cieslinski LB, Sathe GM, Taylor DP, Davis RL, Torphy TJ, Balcarek JM: Cloning and expression of cDNA for a human low-Km, rolipram-sensitive cyclic AMP phosphodiesterase. Mol Cell Biol 10:2678–2686, 1990
  16. Van Lookeren Campagne MM, Wu E, Fleischmann RD, Gottesman MM, Chason KW, Kessin RH: Cyclic AMP responses are suppressed in mammalian cells expressing the yeast low Km cAMP-phosphodiesterase gene. J Biol Chem 265:5840–5846, 1990
  17. Hull RN, Cherry WR, Weaver GW: The origin and characteristics of a pig kidney cell strain, LLC-PK1. In Vitro 12:670–677, 1976 | PubMed | ISI | ChemPort |
  18. Ausiello DA, Hall D: Regulation of vasopressin-sensitive adenylate cyclase by calmodulin. J Biol Chem 256:9796–9798, 1981
  19. Ausiello DA, Hall DH, Dayer J-M: Modulation of cyclic AMP-dependent protein kinase by vasopressin and calcitonin in cultured porcine renal LLC-PK1 cells. Biochem J 186:773–780, 1980
  20. Ishii K, Chang B, Kerwin JF Jr, Wagenaar FL, Huang Z-J, Murad F: Formation of endothelium-derived relaxing factor in porcine kidney epithelial LLC-PK1 cells: An intra- and intercellular messenger for activation of soluble guanylate cyclase. J Pharmacol Exp Ther 256:38–43, 1991 | PubMed | ChemPort |
  21. Leitman DC, Agnost VL, Catalano RM, Schroder H, Waldman SA, Bennett BM, Tuan JJ, Murad F: Atrial natriuretic peptide, oxytocin, and vasopressin increase guanosine 3',5'-monophosphate in LLC-PK1 kidney epithelial cells. Endocrinology 122:1478–1485, 1988
  22. Rassier ME, McIntyre SJ, Yamaki M, Takeda S, Lin J-T, Dousa TP: Isozymes of cyclic-3',5'-nucleotide phosphodiesterases in renal epithelial LLC-PK1 cells. Kidney Int 88–99, 1992 | PubMed | ISI | ChemPort |
  23. Swinnen SV, Joseph DR, Conti M: Molecular cloning of rat homologues of the Drosophila melanogaster dunce cAMP phosphodiesterase: Evidence for a family of genes. Proc Natl Acad Sci USA 86:5325–5329, 1989
  24. Ausubel F, Brent R, Kingston R, Moore D, Seidman J: Introduction of DNA into mammalian cells, in Current Protocols in Molecular Biology, edited by Smith J, Strubel K, New York, John Wiley & Sons, 1990, pp. 901–919
  25. Yamaki M, McIntyre S, Rassier ME, Schwartz JH, Dousa TP: Cyclic 3',5'-nucleotide diesterases in dynamics of cAMP and cGMP in rat collecting duct cells. Am J Physiol 262(R31):F957–F964, 1992 | PubMed | ISI | ChemPort |
  26. Southern PJ, Berg P: Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet 1:327–341, 1982 | PubMed | ChemPort |
  27. Torres VE, Hui YSF, Shah SV, Northrup TE, Dousa TP: Cyclic nucleotide phosphodiesterases in glomeruli of rat renal cortex. Kidney Int 14:444–451, 1978 | PubMed | ISI | ChemPort |
  28. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254, 1976 | Article | PubMed | ISI | ChemPort |
  29. Torres VE, Northrup TE, Edwards RM, Shah SV, Dousa TP: Modulation of cyclic nucleotides in isolated rat glomeruli: Role of histamine, carbamylcholine, parathyroid hormone and angiotensin-II. J Clin Invest 62:1334–1343, 1978
  30. Abboud HE, Shah SV, Dousa TP: Effects of dexamethasone on cyclic nucleotide accumulation in glomeruli. J Lab Clin Med 94:708–717, 1979
  31. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275, 1951 | PubMed | ISI | ChemPort |
  32. Dousa TP, Wilson DM: Effects of demethylchlortetracycline on cellular action of antidiuretic hormone in vitro. Kidney Int 5:279–284, 1974
  33. Chomczyviski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159, 1987
  34. Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995, 1984 | Article | PubMed | ChemPort |
  35. Wells JN, Garat JE, Kramer GL: Inhibition of separated forms of cyclic nucleotide phosphodiesterase from pig coronary arteries by 1,3-disubstituted and 1,3,8-trisubstituted xanthines. J Med Chem 24:954–958, 1981
  36. Kincaid RL, Manganiello VC: Assay of cyclic nucleotide phosphodiesterase using radiolabeled and fluorescent substrates, in Methods in Enzymology, edited by Corbin J, Johnson R, New York, Academic Press, Inc., 1988, p. 457
  37. Swinnen JV, Tsikalas KE, Conti M: Properties and hormonal regulation of two structurally related cAMP phosphodiesterases from the rat Sertoli cell. J Biol Chem 266:18370–18377, 1991 | ChemPort |
  38. Henkel-Tigges J, Davis RL: Rat homologs of the Drosophila dunce gene code for cyclic AMP phosphodiesterases sensitive to rolipram and RO 20-1724. Mol Pharmacol 37:7–10, 1990

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