Pflügers Archiv

, Volume 452, Issue 1, pp 43–52

Evidence that TRPC1 contributes to calcium-induced differentiation of human keratinocytes

  • Shiwei Cai
  • Sahba Fatherazi
  • Richard B. Presland
  • Carol M. Belton
  • Frank A. Roberts
  • Paul C. Goodwin
  • Mark M. Schubert
  • Kenneth T. Izutsu
Epithelial Transport

Abstract

External calcium ion concentration is a major regulator of epidermal keratinocyte differentiation in vitro and probably also in vivo. Regulation of calcium-induced differentiation changes is proposed to occur via an external calcium-sensing, signaling pathway that utilizes increases in intracellular calcium ion concentration to activate differentiation-related gene expression. Calcium ion release from intracellular stores and calcium ion influx via store-operated calcium-permeable channels are key elements in this proposed signaling pathway; however, the channels involved have not yet been identified. The present report shows that human gingival keratinocytes (HGKs) also undergo calcium-induced differentiation in vitro as indicated by involucrin expression and morphological changes. Moreover, TRPC1, which functions as a store-operated calcium channel in a number of cell types, including epidermal keratinocytes, is expressed in both proliferating and differentiating HGKs. Transfection of HGKs with TRPC1 siRNA disrupted expression of TRPC1 mRNA and protein compared with transfection with scrambled TRPC1 siRNA. Cells with disrupted TRPC1 expression showed decreased calcium-induced differentiation as measured by involucrin expression or morphological changes, as well as decreased thapsigargin-induced calcium ion influx, and a decreased rate of store calcium release. These results indicate that TRPC1 is involved in calcium-induced differentiation of HGKs likely by supporting a store-operated calcium ion influx.

Keywords

Differentiation Involucrin Gingiva TRPC Store-operated calcium influx 

References

  1. 1.
    Ahmmed GU, Mehta D, Vogel S, Holinstat M, Paria BC, Tiruppathi C, Malik AB (2004) Protein kinase Calpha phosphorylates the TRPC1 channel and regulates store-operated Ca2+ entry in endothelial cells. J Biol Chem 279:20941–20949CrossRefPubMedGoogle Scholar
  2. 2.
    Beech DJ, Xu SZ, McHugh D, Flemming R (2003) TRPC1 store-operated cationic channel subunit. Cell Calcium 33:433–440CrossRefPubMedGoogle Scholar
  3. 3.
    Belton CM, Izutsu KT, Goodwin PC, Park Y, Lamont RJ (1999) Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells. Cell Microbiol 1:215–223CrossRefPubMedGoogle Scholar
  4. 4.
    Bikle DD, Ng D, Tu CL, Oda Y, Xie Z (2001) Calcium- and vitamin D-regulated keratinocyte differentiation. Mol Cell Endocrinol 177:161–171CrossRefPubMedGoogle Scholar
  5. 5.
    Brown EM, MacLeod RJ (2001) Extracellular calcium sensing and extracellular calcium signaling. Physiol Rev 81:239–297PubMedGoogle Scholar
  6. 6.
    Brown EM, Pollak M, Hebert SC (1995) Molecular mechanisms underlying the sensing of extracellular Ca2+ by parathyroid and kidney cells. Eur J Endocrinol 132:523–531PubMedCrossRefGoogle Scholar
  7. 7.
    Brownlow SL, Harper AG, Harper MT, Sage SO (2004) A role for hTRPC1 and lipid raft domains in store-mediated calcium entry in human platelets. Cell Calcium 35:107–113CrossRefPubMedGoogle Scholar
  8. 8.
    Dolmetsch RE, Lewis RS, Goodnow CC, Healy JI (1997) Differential activation of transcription factors induced by Ca2+ response amplitude and duration. Nature 386:855–858CrossRefPubMedGoogle Scholar
  9. 9.
    Fatherazi S, Belton CM, Cai S, Zarif S, Goodwin PC, Lamont RJ, Izutsu KT (2004) Calcium receptor message, expression and function decrease in differentiating keratinocytes. Pflugers Arch 448:93–104CrossRefPubMedGoogle Scholar
  10. 10.
    Fatherazi S, Belton CM, Izutsu KT (2003) Sequential activation of store-operated currents in human gingival keratinocytes. J Invest Dermatol 121:120–131CrossRefPubMedGoogle Scholar
  11. 11.
    Forslind B, Lindberg M, Roomans GM, Pallon J, Werner-Linde Y (1997) Aspects on the physiology of human skin: studies using particle probe analysis. Microsc Res Tech 38:373–386CrossRefPubMedGoogle Scholar
  12. 12.
    Gibson DF, Ratnam AV, Bikle DD (1996) Evidence for separate control mechanisms at the message, protein, and enzyme activation levels for transglutaminase during calcium-induced differentiation of normal and transformed human keratinocytes. J Invest Dermatol 106:154–161CrossRefPubMedGoogle Scholar
  13. 13.
    Golovina VA (2005) Visualization of localized store-operated calcium entry in mouse astrocytes. Close proximity to the endoplasmic reticulum. J Physiol 564(3):737–749CrossRefPubMedGoogle Scholar
  14. 14.
    Jones KT, Sharpe GR (1994) Thapsigargin raises intracellular free calcium levels in human keratinocytes and inhibits the coordinated expression of differentiation markers. Exp Cell Res 210:71–76CrossRefPubMedGoogle Scholar
  15. 15.
    Komuves L, Oda Y, Tu CL, Chang WH, Ho-Pao CL, Mauro T, Bikle DD (2002) Epidermal expression of the full-length extracellular calcium-sensing receptor is required for normal keratinocyte differentiation. J Cell Physiol 192:45–54CrossRefPubMedGoogle Scholar
  16. 16.
    Kunzelmann-Marche C, Freyssinet JM, Martinez MC (2002) Loss of plasma membrane phospholipid asymmetry requires raft integrity. Role of transient receptor potential channels and ERK pathway. J Biol Chem 277:19876–19881CrossRefPubMedGoogle Scholar
  17. 17.
    La Celle PT, Polakowska RR (2001) Human homeobox HOXA7 regulates keratinocyte transglutaminase type 1 and inhibits differentiation. J Biol Chem 276:32844–32853CrossRefPubMedGoogle Scholar
  18. 18.
    Li L, Tucker RW, Hennings H, Yuspa SH (1995) Chelation of intracellular Ca2+ inhibits murine keratinocyte differentiation in vitro. J Cell Physiol 163:105–114CrossRefPubMedGoogle Scholar
  19. 19.
    Liu X, Singh BB, Ambudkar IS (2003) TRPC1 is required for functional store-operated Ca2+ channels. Role of acidic amino acid residues in the S5–S6 region. J Biol Chem 278:11337–11343CrossRefPubMedGoogle Scholar
  20. 20.
    Mauro T, Bench G, Sidderas-Haddad E, Feingold K, Elias P, Cullander C (1998) Acute barrier perturbation abolishes the Ca2+ and K+ gradients in murine epidermis: quantitative measurement using PIXE. J Invest Dermatol 111:1198–1201CrossRefPubMedGoogle Scholar
  21. 21.
    McKay RR, Szymeczek-Seay CL, Lievremont JP, Bird GS, Zitt C, Jungling E, Luckhoff A, Putney JW Jr (2000) Cloning and expression of the human transient receptor potential 4 (TRP4) gene: localization and functional expression of human TRP4 and TRP3. Biochem J 351(Pt 3):735–746CrossRefPubMedGoogle Scholar
  22. 22.
    Milewski MI, Mickle JE, Forrest JK, Stanton BA, Cutting GR (2002) Aggregation of misfolded proteins can be a selective process dependent upon peptide composition. J Biol Chem 277:34462–34470CrossRefPubMedGoogle Scholar
  23. 23.
    Mori Y, Wakamori M, Miyakawa T, Hermosura M, Hara Y, Nishida M, Hirose K, Mizushima A, Kurosaki M, Mori E, Gotoh K, Okada T, Fleig A, Penner R, Iino M, Kurosaki T (2002) Transient receptor potential 1 regulates capacitative Ca(2+) entry and Ca(2+) release from endoplasmic reticulum in B lymphocytes. J Exp Med 195:673–681CrossRefPubMedGoogle Scholar
  24. 24.
    Presland RB, Dale BA (2000) Epithelial structural proteins of the skin and oral cavity: function in health and disease. Crit Rev Oral Biol Med 11:383–408PubMedGoogle Scholar
  25. 25.
    Presland RB, Kimball JR, Kautsky MB, Lewis SP, Lo CY, Dale BA (1997) Evidence for specific proteolytic cleavage of the N-terminal domain of human profilaggrin during epidermal differentiation. J Invest Dermatol 108:170–178CrossRefPubMedGoogle Scholar
  26. 26.
    Rosado JA, Brownlow SL, Sage SO (2002) Endogenously expressed Trp1 is involved in store-mediated Ca2 entry by conformational coupling in human platelets. J Biol Chem 277:42157–42163CrossRefPubMedGoogle Scholar
  27. 27.
    Rosado JA, Sage SO (2001) Activation of store-mediated calcium entry by secretion-like coupling between the inositol 1,4,5-trisphosphate receptor type II and human transient receptor potential (hTrp1) channels in human platelets. Biochem J 356:191–198CrossRefPubMedGoogle Scholar
  28. 28.
    Sharpe GR, Gillespie JI, Greenwell JR (1989) An increase in intracellular free calcium is an early event during differentiation of cultured human keratinocytes. FEBS Lett 254:25–28CrossRefPubMedGoogle Scholar
  29. 29.
    Spassova MA, Soboloff J, He LP, Hewavitharana T, Xu W, Venkatachalam K, van Rossum DB, Patterson RL, Gill DL (2004) Calcium entry mediated by SOCs and TRP channels: variations and enigma. Biochim Biophys Acta 1742:9–20CrossRefPubMedGoogle Scholar
  30. 30.
    Strubing C, Krapivinsky G, Krapivinsky L, Clapham DE (2001) TRPC1 and TRPC5 form a novel cation channel in mammalian brain. Neuron 29:645–655CrossRefPubMedGoogle Scholar
  31. 31.
    Sumitomo S, Kumasa S, Iwai Y, Mori M (1986) Involucrin expression in epithelial tumors of oral and pharyngeal mucosa and skin. Oral Surg Oral Med Oral Pathol 62:155–163CrossRefPubMedGoogle Scholar
  32. 32.
    Sweeney M, Yu Y, Platoshyn O, Zhang S, McDaniel SS, Yuan JX (2002) Inhibition of endogenous TRP1 decreases capacitative Ca2+ entry and attenuates pulmonary artery smooth muscle cell proliferation. Am J Physiol Lung Cell Mol Physiol 283:L144–155PubMedGoogle Scholar
  33. 33.
    Tu CL, Chang W, Bikle DD (2005) Phospholipase cgamma1 is required for activation of store-operated channels in human keratinocytes. J Invest Dermatol 124:187–197CrossRefPubMedGoogle Scholar
  34. 34.
    Vanden Abeele F, Lemonnier L, Thebault S, Lepage G, Parys JB, Shuba Y, Skryma R, Prevarskaya N (2004) Two types of store-operated Ca2 channels with different activation modes and molecular origin in LNCaP human prostate cancer epithelial cells. J Biol Chem 279:30326–30337CrossRefPubMedGoogle Scholar
  35. 35.
    Wang W, O'Connell B, Dykeman R, Sakai T, Delporte C, Swaim W, Zhu X, Birnbaumer L, Ambudkar IS (1999) Cloning of Trp1beta isoform from rat brain: immunodetection and localization of the endogenous Trp1 protein. Am J Physiol 276:C969–979PubMedGoogle Scholar
  36. 36.
    Watt FM, Mattey DL, Garrod DR (1984) Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol 99:2211–2215CrossRefPubMedGoogle Scholar
  37. 37.
    Wu X, Babnigg G, Zagranichnaya T, Villereal ML (2002) The role of endogenous human Trp4 in regulating carbachol-induced calcium oscillations in HEK-293 cells. J Biol Chem 277:13597–13608CrossRefPubMedGoogle Scholar
  38. 38.
    Wu X, Zagranichnaya TK, Gurda GT, Eves EM, Villereal ML (2004) A TRPC1/TRPC3-mediated increase in store-operated calcium entry is required for differentiation of H19-7 hippocampal neuronal cells. J Biol Chem 279:43392–43402CrossRefPubMedGoogle Scholar
  39. 39.
    Xu XZ, Li HS, Guggino WB, Montell C (1997) Coassembly of TRP and TRPL produces a distinct store-operated conductance. Cell 89:1155–1164CrossRefPubMedGoogle Scholar
  40. 40.
    Yuspa SH, Hennings H, Tucker RW, Jaken S, Kilkenny AE, Roop DR (1988) Signal transduction for proliferation and differentiation in keratinocytes. Ann N Y Acad Sci 548:191–196PubMedCrossRefGoogle Scholar
  41. 41.
    Cai S, Fatherazi S, Presland RB, Belton CM, Iźutsu KT (2005) TRPC channel expression during calcium-induced differentiation of human gingival keratinocytes. J Dermatol Sci 40:21–28CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Shiwei Cai
    • 1
  • Sahba Fatherazi
    • 1
  • Richard B. Presland
    • 1
    • 2
  • Carol M. Belton
    • 1
  • Frank A. Roberts
    • 3
  • Paul C. Goodwin
    • 4
  • Mark M. Schubert
    • 5
  • Kenneth T. Izutsu
    • 1
  1. 1.Department of Oral BiologyUniversity of WashingtonSeattleUSA
  2. 2.Division of Dermatology, Department of MedicineUniversity of WashingtonSeattleUSA
  3. 3.Department of PeriodonticsUniversity of WashingtonSeattleUSA
  4. 4.Applied PrecisionIssaquahUSA
  5. 5.Department of Oral MedicineUniversity of WashingtonSeattleUSA

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