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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Mar;82(6):1736–1740. doi: 10.1073/pnas.82.6.1736

Stimulation of Na+/H+ antiport is an early event in hypertrophy of renal proximal tubular cells.

L G Fine, B Badie-Dezfooly, A G Lowe, A Hamzeh, J Wells, S Salehmoghaddam
PMCID: PMC397347  PMID: 3885217

Abstract

Renal hypertrophy in vivo is achieved by an increase in protein content per cell and an increase in cell size with minimal hyperplasia. Hypertrophied renal tubular cells remain quiescent and demonstrate an increase in transcellular transport rates. This situation was simulated in vitro by exposing a confluent, quiescent primary culture of rabbit renal proximal tubular cells to either insulin, prostaglandin E1, or hypertonic NaCl for 24 or 48 hr. Protein per cell increased by 20-30% with little or no increase in [3H]thymidine incorporation into DNA. Mean cell volume was also increased in insulin- and hypertonic NaCl-treated but not in prostaglandin E1-treated cells. The lag period required to initiate DNA synthesis by a combination of insulin and hydrocortisone was the same in control and hypertrophied cells, indicating a quiescent state of the latter. Two hours of exposure to the growth stimuli increased amiloride-sensitive Na+ uptake, Na-dependent H+ efflux, and ouabain-sensitive Rb+ uptake, indicating that stimulation of Na+/H+ antiport (exchange) occurs as an early event in their action. Hypertrophied cells continued to demonstrate enhanced Na+/H+ antiport after the growth stimuli were removed for 3 hr, by which time their acute effects are reversed.

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Selected References

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  1. Baralle F. E., Shoulders C. C., Proudfoot N. J. The primary structure of the human epsilon-globin gene. Cell. 1980 Oct;21(3):621–626. doi: 10.1016/0092-8674(80)90425-0. [DOI] [PubMed] [Google Scholar]
  2. Bell G. I., Pictet R. L., Rutter W. J., Cordell B., Tischer E., Goodman H. M. Sequence of the human insulin gene. Nature. 1980 Mar 6;284(5751):26–32. doi: 10.1038/284026a0. [DOI] [PubMed] [Google Scholar]
  3. Besterman J. M., Tyrey S. J., Cragoe E. J., Jr, Cuatrecasas P. Inhibition of epidermal growth factor-induced mitogenesis by amiloride and an analog: evidence against a requirement for Na+/H+ exchange. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6762–6766. doi: 10.1073/pnas.81.21.6762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang A. C., Cochet M., Cohen S. N. Structural organization of human genomic DNA encoding the pro-opiomelanocortin peptide. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4890–4894. doi: 10.1073/pnas.77.8.4890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Czelusniak J., Goodman M., Hewett-Emmett D., Weiss M. L., Venta P. J., Tashian R. E. Phylogenetic origins and adaptive evolution of avian and mammalian haemoglobin genes. Nature. 1982 Jul 15;298(5871):297–300. doi: 10.1038/298297a0. [DOI] [PubMed] [Google Scholar]
  6. Efstratiadis A., Posakony J. W., Maniatis T., Lawn R. M., O'Connell C., Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M., Slightom J. L. The structure and evolution of the human beta-globin gene family. Cell. 1980 Oct;21(3):653–668. doi: 10.1016/0092-8674(80)90429-8. [DOI] [PubMed] [Google Scholar]
  7. Ellison J. W., Berson B. J., Hood L. E. The nucleotide sequence of a human immunoglobulin C gamma1 gene. Nucleic Acids Res. 1982 Jul 10;10(13):4071–4079. doi: 10.1093/nar/10.13.4071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Emorine L., Dreher K., Kindt T. J., Max E. E. Rabbit immunoglobulin kappa genes: structure of a germline b4 allotype J-C locus and evidence for several b4-related sequences in the rabbit genome. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5709–5713. doi: 10.1073/pnas.80.18.5709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Erlinger S. U., Saier M. H., Jr Decrease in protein content and cell volume of cultured dog kidney epithelial cells during growth. In Vitro. 1982 Mar;18(3 Pt 1):196–202. doi: 10.1007/BF02618571. [DOI] [PubMed] [Google Scholar]
  10. Fiddes J. C., Goodman H. M. The gene encoding the common alpha subunit of the four human glycoprotein hormones. J Mol Appl Genet. 1981;1(1):3–18. [PubMed] [Google Scholar]
  11. Fine L. G. Adaptation of renal tubule in uremia. Kidney Int. 1982 Nov;22(5):546–549. doi: 10.1038/ki.1982.208. [DOI] [PubMed] [Google Scholar]
  12. Fine L. G., Trizna W., Bourgoignie J. J., Bricker N. S. Functional profile of the isolated uremic nephron. Role of compensatory hypertrophy in the control of fluid reabsorption by the proximal straight tubule. J Clin Invest. 1978 Jun;61(6):1508–1518. doi: 10.1172/JCI109071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fitch W. M. Estimating the total number of nucleotide substitutions since the common ancestor of a pair of homologous genes: comparison of several methods and three beta hemoglobin messenger RNA's. J Mol Evol. 1980 Dec;16(3-4):153–209. doi: 10.1007/BF01804976. [DOI] [PubMed] [Google Scholar]
  14. Godine J. E., Chin W. W., Habener J. F. alpha Subunit of rat pituitary glycoprotein hormones. Primary structure of the precursor determined from the nucleotide sequence of cloned cDNAs. J Biol Chem. 1982 Jul 25;257(14):8368–8371. [PubMed] [Google Scholar]
  15. Haley J., Hudson P., Scanlon D., John M., Cronk M., Shine J., Tregear G., Niall H. Porcine relaxin: molecular cloning and cDNA structure. DNA. 1982;1(2):155–162. doi: 10.1089/dna.1.1982.1.155. [DOI] [PubMed] [Google Scholar]
  16. Hanauer A., Levin M., Heilig R., Daegelen D., Kahn A., Mandel J. L. Isolation and characterization of cDNA clones for human skeletal muscle alpha actin. Nucleic Acids Res. 1983 Jun 11;11(11):3503–3516. doi: 10.1093/nar/11.11.3503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hansen J. N., Konkel D. A., Leder P. The sequence of a mouse embryonic beta-globin gene. Evolution of the gene and its signal region. J Biol Chem. 1982 Jan 25;257(2):1048–1052. [PubMed] [Google Scholar]
  18. Hardison R. C. Comparison of the beta-like globin gene families of rabbits and humans indicates that the gene cluster 5'-epsilon-gamma-delta-beta-3' predates the mammalian radiation. Mol Biol Evol. 1984 Sep;1(5):390–410. doi: 10.1093/oxfordjournals.molbev.a040326. [DOI] [PubMed] [Google Scholar]
  19. Heidmann O., Rougeon F. Molecular cloning of rabbit gamma heavy chain mRNA. Nucleic Acids Res. 1982 Mar 11;10(5):1535–1545. doi: 10.1093/nar/10.5.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Heinrich G., Kronenberg H. M., Potts J. T., Jr, Habener J. F. Gene encoding parathyroid hormone. Nucleotide sequence of the rat gene and deduced amino acid sequence of rat preproparathyroid hormone. J Biol Chem. 1984 Mar 10;259(5):3320–3329. [PubMed] [Google Scholar]
  21. Hendy G. N., Kronenberg H. M., Potts J. T., Jr, Rich A. Nucleotide sequence of cloned cDNAs encoding human preproparathyroid hormone. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7365–7369. doi: 10.1073/pnas.78.12.7365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hieter P. A., Max E. E., Seidman J. G., Maizel J. V., Jr, Leder P. Cloned human and mouse kappa immunoglobulin constant and J region genes conserve homology in functional segments. Cell. 1980 Nov;22(1 Pt 1):197–207. doi: 10.1016/0092-8674(80)90168-3. [DOI] [PubMed] [Google Scholar]
  23. Hill A., Hardies S. C., Phillips S. J., Davis M. G., Hutchison C. A., 3rd, Edgell M. H. Two mouse early embryonic beta-globin gene sequences. Evolution of the nonadult beta-globins. J Biol Chem. 1984 Mar 25;259(6):3739–3747. [PubMed] [Google Scholar]
  24. Honjo T., Obata M., Yamawaki-Katoaka Y., Kataoka T., Kawakami T., Takahashi N., Mano Y. Cloning and complete nucleotide sequence of mouse immunoglobulin gamma 1 chain gene. Cell. 1979 Oct;18(2):559–568. doi: 10.1016/0092-8674(79)90072-2. [DOI] [PubMed] [Google Scholar]
  25. Hudson P., Haley J., Cronk M., Shine J., Niall H. Molecular cloning and characterization of cDNA sequences coding for rat relaxin. Nature. 1981 May 14;291(5811):127–131. doi: 10.1038/291127a0. [DOI] [PubMed] [Google Scholar]
  26. Hudson P., Haley J., John M., Cronk M., Crawford R., Haralambidis J., Tregear G., Shine J., Niall H. Structure of a genomic clone encoding biologically active human relaxin. Nature. 1983 Feb 17;301(5901):628–631. doi: 10.1038/301628a0. [DOI] [PubMed] [Google Scholar]
  27. Johnson H. A., Vera Roman J. M. Compensatory renal enlargement. Hypertrophy versus hyperplasia. Am J Pathol. 1966 Jul;49(1):1–13. [PMC free article] [PubMed] [Google Scholar]
  28. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980 Dec;16(2):111–120. doi: 10.1007/BF01731581. [DOI] [PubMed] [Google Scholar]
  29. Konkel D. A., Maizel J. V., Jr, Leder P. The evolution and sequence comparison of two recently diverged mouse chromosomal beta--globin genes. Cell. 1979 Nov;18(3):865–873. doi: 10.1016/0092-8674(79)90138-7. [DOI] [PubMed] [Google Scholar]
  30. Kwok S. C., Chan S. J., Steiner D. F. Cloning and nucleotide sequence analysis of the dog insulin gene. Coded amino acid sequence of canine preproinsulin predicts an additional C-peptide fragment. J Biol Chem. 1983 Feb 25;258(4):2357–2363. [PubMed] [Google Scholar]
  31. L'Allemain G., Franchi A., Cragoe E., Jr, Pouysségur J. Blockade of the Na+/H+ antiport abolishes growth factor-induced DNA synthesis in fibroblasts. Structure-activity relationships in the amiloride series. J Biol Chem. 1984 Apr 10;259(7):4313–4319. [PubMed] [Google Scholar]
  32. L'Allemain G., Paris S., Pouysségur J. Growth factor action and intracellular pH regulation in fibroblasts. Evidence for a major role of the Na+/H+ antiport. J Biol Chem. 1984 May 10;259(9):5809–5815. [PubMed] [Google Scholar]
  33. Laird C. D., McConaughy B. L., McCarthy B. J. Rate of fixation of nucleotide substitutions in evolution. Nature. 1969 Oct 11;224(5215):149–154. doi: 10.1038/224149a0. [DOI] [PubMed] [Google Scholar]
  34. Lawn R. M., Efstratiadis A., O'Connell C., Maniatis T. The nucleotide sequence of the human beta-globin gene. Cell. 1980 Oct;21(3):647–651. doi: 10.1016/0092-8674(80)90428-6. [DOI] [PubMed] [Google Scholar]
  35. Leffert H. L., Koch K. S., Fehlmann M., Heiser W., Lad P. J., Skelly H. Amiloride blocks cell-free protein synthesis at levels attained inside cultured rat hepatocytes. Biochem Biophys Res Commun. 1982 Sep 30;108(2):738–745. doi: 10.1016/0006-291x(82)90891-9. [DOI] [PubMed] [Google Scholar]
  36. Liebhaber S. A., Goossens M. J., Kan Y. W. Cloning and complete nucleotide sequence of human 5'-alpha-globin gene. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7054–7058. doi: 10.1073/pnas.77.12.7054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lomedico P., Rosenthal N., Efstratidadis A., Gilbert W., Kolodner R., Tizard R. The structure and evolution of the two nonallelic rat preproinsulin genes. Cell. 1979 Oct;18(2):545–558. doi: 10.1016/0092-8674(79)90071-0. [DOI] [PubMed] [Google Scholar]
  38. Mendoza S. A., Wigglesworth N. M., Rozengurt E. Vasopressin rapidly stimulates Na entry and Na-K pump activity in quiescent cultures of mouse 3T3 cells. J Cell Physiol. 1980 Oct;105(1):153–162. doi: 10.1002/jcp.1041050117. [DOI] [PubMed] [Google Scholar]
  39. Miller W. L., Eberhardt N. L. Structure and evolution of the growth hormone gene family. Endocr Rev. 1983 Spring;4(2):97–130. doi: 10.1210/edrv-4-2-97. [DOI] [PubMed] [Google Scholar]
  40. Miyata T., Yasunaga T., Nishida T. Nucleotide sequence divergence and functional constraint in mRNA evolution. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7328–7332. doi: 10.1073/pnas.77.12.7328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  42. Moolenaar W. H., Yarden Y., de Laat S. W., Schlessinger J. Epidermal growth factor induces electrically silent Na+ influx in human fibroblasts. J Biol Chem. 1982 Jul 25;257(14):8502–8506. [PubMed] [Google Scholar]
  43. Nakanishi S., Teranishi Y., Watanabe Y., Notake M., Noda M., Kakidani H., Jingami H., Numa S. Isolation and characterization of the bovine corticotropin/beta-lipotropin precursor gene. Eur J Biochem. 1981 Apr;115(3):429–438. doi: 10.1111/j.1432-1033.1981.tb06220.x. [DOI] [PubMed] [Google Scholar]
  44. Nilson J. H., Thomason A. R., Cserbak M. T., Moncman C. L., Woychik R. P. Nucleotide sequence of a cDNA for the common alpha subunit of the bovine pituitary glycoprotein hormones. Conservation of nucleotides in the 3'-untranslated region of bovine and human pre-alpha subunit mRNAs. J Biol Chem. 1983 Apr 25;258(8):4679–4682. [PubMed] [Google Scholar]
  45. Nishioka Y., Leder P. The complete sequence of a chromosomal mouse alpha--globin gene reveals elements conserved throughout vertebrate evolution. Cell. 1979 Nov;18(3):875–882. doi: 10.1016/0092-8674(79)90139-9. [DOI] [PubMed] [Google Scholar]
  46. Nomura K., Puett D., Nicholson W. E., Liddle G. W. Partial purification and characterization of a renotropic fraction from ovine pituitaries. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6675–6679. doi: 10.1073/pnas.79.21.6675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Notake M., Tobimatsu T., Watanabe Y., Takahashi H., Mishina M., Numa S. Isolation and characterization of the mouse corticotropin-beta-lipotropin precursor gene and a related pseudogene. FEBS Lett. 1983 May 30;156(1):67–71. doi: 10.1016/0014-5793(83)80250-6. [DOI] [PubMed] [Google Scholar]
  48. Owen N. E., Villereal M. L. Lys-bradykinin stimulates Na+ influx and DNA synthesis in cultured human fibroblasts. Cell. 1983 Mar;32(3):979–985. doi: 10.1016/0092-8674(83)90082-x. [DOI] [PubMed] [Google Scholar]
  49. Pouysségur J., Chambard J. C., Franchi A., Paris S., Van Obberghen-Schilling E. Growth factor activation of an amiloride-sensitive Na+/H+ exchange system in quiescent fibroblasts: coupling to ribosomal protein S6 phosphorylation. Proc Natl Acad Sci U S A. 1982 Jul;79(13):3935–3939. doi: 10.1073/pnas.79.13.3935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Putney S. D., Herlihy W. C., Schimmel P. A new troponin T and cDNA clones for 13 different muscle proteins, found by shotgun sequencing. Nature. 1983 Apr 21;302(5910):718–721. doi: 10.1038/302718a0. [DOI] [PubMed] [Google Scholar]
  51. Rothenberg P., Glaser L., Schlesinger P., Cassel D. Epidermal growth factor stimulates amiloride-sensitive 22Na+ uptake in A431 cells. Evidence for Na+/H+ exchange. J Biol Chem. 1983 Apr 25;258(8):4883–4889. [PubMed] [Google Scholar]
  52. Rothenberg P., Reuss L., Glaser L. Serum and epidermal growth factor transiently depolarize quiescent BSC-1 epithelial cells. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7783–7787. doi: 10.1073/pnas.79.24.7783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sarich V. M., Wilson A. C. Immunological time scale for hominid evolution. Science. 1967 Dec 1;158(3805):1200–1203. doi: 10.1126/science.158.3805.1200. [DOI] [PubMed] [Google Scholar]
  54. Schon E. A., Cleary M. L., Haynes J. R., Lingrel J. B. Structure and evolution of goat gamma-, beta C- and beta A-globin genes: three developmentally regulated genes contain inserted elements. Cell. 1981 Dec;27(2 Pt 1):359–369. doi: 10.1016/0092-8674(81)90419-0. [DOI] [PubMed] [Google Scholar]
  55. Schon E. A., Wernke S. M., Lingrel J. B. Gene conversion of two functional goat alpha-globin genes preserves only minimal flanking sequences. J Biol Chem. 1982 Jun 25;257(12):6825–6835. [PubMed] [Google Scholar]
  56. Schuldiner S., Rozengurt E. Na+/H+ antiport in Swiss 3T3 cells: mitogenic stimulation leads to cytoplasmic alkalinization. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7778–7782. doi: 10.1073/pnas.79.24.7778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Sedmak J. J., Grossberg S. E. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. doi: 10.1016/0003-2697(77)90428-6. [DOI] [PubMed] [Google Scholar]
  58. Slightom J. L., Blechl A. E., Smithies O. Human fetal G gamma- and A gamma-globin genes: complete nucleotide sequences suggest that DNA can be exchanged between these duplicated genes. Cell. 1980 Oct;21(3):627–638. doi: 10.1016/0092-8674(80)90426-2. [DOI] [PubMed] [Google Scholar]
  59. Soltoff S. P., Mandel L. J. Amiloride directly inhibits the Na,K-ATPase activity of rabbit kidney proximal tubules. Science. 1983 May 27;220(4600):957–958. doi: 10.1126/science.6302840. [DOI] [PubMed] [Google Scholar]
  60. Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M. Complete nucleotide sequence of the human delta-globin gene. Cell. 1980 Oct;21(3):639–646. doi: 10.1016/0092-8674(80)90427-4. [DOI] [PubMed] [Google Scholar]
  61. Tabei K., Levenson D. J., Brenner B. M. Early enhancement of fluid transport in rabbit proximal straight tubules after loss of contralateral renal excretory function. J Clin Invest. 1983 Sep;72(3):871–881. doi: 10.1172/JCI111058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Takahashi H., Hakamata Y., Watanabe Y., Kikuno R., Miyata T., Numa S. Complete nucleotide sequence of the human corticotropin-beta-lipotropin precursor gene. Nucleic Acids Res. 1983 Oct 11;11(19):6847–6858. doi: 10.1093/nar/11.19.6847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Trizna W., Yanagawa N., Bar-Khayim Y., Houston B., Fine L. G. Functional profile of the isolated uremic nephron. Evidence of proximal tubular "memory" in experimental renal disease. J Clin Invest. 1981 Sep;68(3):760–767. doi: 10.1172/JCI110312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Uhler M., Herbert E. Complete amino acid sequence of mouse pro-opiomelanocortin derived from the nucleotide sequence of pro-opiomelanocortin cDNA. J Biol Chem. 1983 Jan 10;258(1):257–261. [PubMed] [Google Scholar]
  65. Wilson A. C., Carlson S. S., White T. J. Biochemical evolution. Annu Rev Biochem. 1977;46:573–639. doi: 10.1146/annurev.bi.46.070177.003041. [DOI] [PubMed] [Google Scholar]
  66. Winter R. M., Tuddenham E. G., Goldman E., Matthews K. B. A maximum likelihood estimate of the sex ratio of mutation rates in haemophilia A. Hum Genet. 1983;64(2):156–159. doi: 10.1007/BF00327115. [DOI] [PubMed] [Google Scholar]
  67. Zakut R., Shani M., Givol D., Neuman S., Yaffe D., Nudel U. Nucleotide sequence of the rat skeletal muscle actin gene. Nature. 1982 Aug 26;298(5877):857–859. doi: 10.1038/298857a0. [DOI] [PubMed] [Google Scholar]

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