Skip to main content
Log in

Parathyroid hormone-induced changes of the brush border topography and cytoskeleton in cultured renal proximal tubular cells

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

In order to examine the possibility of parathyroid hormone-mediated ultrastructural rearrangements in target epithelium, isolated canine renal proximal tubular cells were grown on a collagen-coated semipermeable membrane in a defined medium. Scanning and transmission electron microscopy of these monolayers revealed abundant microvilli. Exposure of the proximal tubular cells to parathyroid hormone resulted in a biphasic changes involving: (1) dramatic shortening and rarefaction of microvilli within 1 min; and (2) recovery of microvillar topography after 5 min. A similar shortening of microvilli was observed following exposure to ionomycin, whereas incubation with cyclic AMP resulted in an elongation of microvilli. Parathyroid hormone stimulated cyclic AMP production and increased cytoplasmic free calcium concentration in cultured proximal tubular cells. Pretreatment of cells with a calmodulin inhibitor abolished the effect of parathyroid hormone on brush border topography. Shortening of microvilli was associated with a disappearance of microvillar core filaments. Staining of F-actin with fluoresceinphalloidin showed that parathyroid hormone resulted in fragmentation of stress fibers. It is concluded that parathyroid hormoneinduced cell activation involves cytoplasmic-free calcium, calmodulin, and the cytoskeleton.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Ausiello, D.A., Bonventre, J.V. 1984. Calcium and calmodulin as mediators of hormone action and transport events.Semin. Nephrol. 4:134–143

    Google Scholar 

  2. Borle, A., Uchikawa, T. 1987. Effects of parathyroid hormone on the distribution and transport of calcium in cultured kidney cells.Endocrinology 102:1725–1732

    Google Scholar 

  3. Bretscher, A. 1983. Molecular architecture of the microvillus cytoskeleton.In: Brush Border Membranes. Ciba Foundation Symposium, 95. pp. 164–179, Pitman Books, London

    Google Scholar 

  4. Burgess, D.R., Prum, B.E. 1982. Re-evaluation of brush border motility: Calcium induces core filament solution and microvillar vesiculation.J. Cell Biol. 94:97–107

    PubMed  Google Scholar 

  5. Chao, W.T.H., Forte, L.R. 1982. Rat kidney cells in primary culture: Hormone-mediated desensitization of the adenosine 3′, 5′-monophosphate response to parathyroid hormone and calcitonin.Endocrinology 111:252–259

    PubMed  Google Scholar 

  6. Chase, H.S. 1984. Does calcium couple the apical and basolateral membrane permeabilities in epithelia?Am. J. Physiol. 247:F869-F876

    Google Scholar 

  7. Chou, K., Yamashita, N., Kurokawa, K. 1985. Membrane potential changes in cloned monkey proximal tubular cells: Effect of parathyroid hormone.Clin. Res. 33:583A (abstr.)

    Google Scholar 

  8. Cohn, D.E., Klahr, S., Hammerman, M.R. 1983. Metabolic acidosis and parathyroidectomy increase Na+/H+ exchange in brush border vesicles.Am. J. Physiol. 245:F217-F222

    PubMed  Google Scholar 

  9. Cook, N.S., Haylett, D.G. 1985. Effects of apamin, quinine and neuromuscular blockers on calcium-activated potassium channels in guinea-pig hepatocytes.J. Physiol. (London) 358:373–394

    Google Scholar 

  10. Dennis, V., Bello-Reuss, E., Robinson, R. 1977. Response of phosphate transport to parathyroid hormone in segments of rabbit nephron.Am. J. Physiol. 233:F29-F38

    PubMed  Google Scholar 

  11. Diamond, J. 1983. Transcellular cross talk between epithelial cell membranes.Nature (London) 300:683–685

    Google Scholar 

  12. DiBona, D.R. 1983. Cytoplasmic involvement in ADH-mediated osmosis across toad urinary bladder.Am. J. Physiol. 245:C297-C307

    PubMed  Google Scholar 

  13. Dolson, G.M., Hise, M.K., Weinman, E.J. 1985. Relationship among parathyroid hormone, cAMP, and calcium on proximal tubule sodium transport.Am. J. Physiol. 249:F409-F416

    PubMed  Google Scholar 

  14. Glenney, J.R., Glenney, P. 1985. Comparison of Ca2+-regulated events in the intestinal brush border.J. Cell Biol. 100:754–763

    PubMed  Google Scholar 

  15. Grinstein, A., Dupre, A., Rothstein, A. 1982. Volume regulation by human lymphocytes. Role of calcium.J. Gen. Physiol. 79:849–868

    Google Scholar 

  16. Hammerman, M.R., Hruska, K.A. 1982. Cyclic-AMP-dependent protein phosphorylation in canine renal brush border membrane vesicles is associated with decreased phosphate transport.J. Biol. Chem. 257:992–999

    PubMed  Google Scholar 

  17. Harmanci, M.C., Kachadorian, W.A., Valtin, H., DiScala, V.A. 1978. Antidiuretic hormone-induced intramembranous alterations in mammalian collecting ducts.Am. J. Physiol. 235:F440-F443

    Google Scholar 

  18. Harper, J.F., Brooker, N. 1985. Femtomole sensitive radioimmunoassay for cyclic AMP and cyclic GMP after 2′0 acetylation by acetic anhydrate in aqueous solutions.J. Cyclic Nucleotide Res. 1:207–215

    Google Scholar 

  19. Hays, R.M. 1983. Alteration of luminal membrane structure by antidiuretic hormone.Am. J. Physiol. 245:C289-C296

    PubMed  Google Scholar 

  20. Henry, H.L., Cunningham, N.S., Noland, T.A. 1983. Homologous desensitization of cultured chick kidney cells to parathyroid hormone.Endocrinology 113:1942–1949

    PubMed  Google Scholar 

  21. Hruska, K., Goligorsky, M., Scoble, J., Tsutsumi, S., Westbrook, S., Moskowitz, D. 1986. The effects of parathyroid hormone on cytosolic calcium in renal proximal tubular primary cultures.Am. J. Physiol. (in press)

  22. Hruska, K., Khalifa, S., Meltzer, V., Martin, K. 1984. Calcium as a modulator of the physiologic and pathophysiologic effects of parathyroid hormone.Semin. Nephrol. 4:159–173

    Google Scholar 

  23. Jayakumar, A., Cheng, L., Liang, C.T., Sacktor, B. 1984. Sodium gradient-dependent calcium uptake in renal basolateral membrane vesicles. Effect of parathyroid hormone.J. Biol. Chem. 259(17):10827–10833

    PubMed  Google Scholar 

  24. Jo, O., Yanagawa, N. 1984. Trifluoperazin abolishes parathyroid hormone effect on phosphate uptake by isolated rabbit renal brush border membrane. Am. Soc. Nephrol. 17th Meeting, 20A (abstr).

  25. Kahn, A.M., Dolson, G.M., Hise, M.K., Bennett, S.C., Weinman, E.J. 1985. Parathyroid hormone and dibutyryl cAMP inhibit Na+/H+ exchange in renal brush border vesicles.Am. J. Physiol. 248:F212-F218

    Google Scholar 

  26. Keller, T.C.S., Mooseker, M.S. 1982. Ca2+-calmodulin-dependent phosphorylation of myosin, and its role in brush border contractionin vitro.J. Cell. Biol. 95:943–959

    PubMed  Google Scholar 

  27. Khalifa, S., Mills, S., Hruska, K.A. 1983. Stimulation of calcium uptake by parathyroid hormone in renal brush border membrane vesicles: Relationship to membrane phosphorylation.J. Biol. Chem. 258:14400–14406

    PubMed  Google Scholar 

  28. Larson, M., Spring, K.R. 1984. Volume regulation byNecturus gallbladder: Basolateral KCl exit.J. Membrane Biol. 81:219–232

    Google Scholar 

  29. LeFurgey, A., Dratwa, M., Tisher, C.C. 1981. Effects of colchicine and cytochalasin B on vasopressin and cyclic adenosine monophosphate-induced changes in toad urinary bladder.Lab. Invest. 45:308–315

    PubMed  Google Scholar 

  30. LeGrimellec, C., Giocondi, B., Carriere, M.C., Cardinal, J. 1982. Membrane fluidity and enzyme activities in brush border and basolateral membranes of the dog kidney.Am. J. Physiol. 242:F246-F253

    Google Scholar 

  31. Marcus, R., Grant, B.F. 1983. Parathyroid hormone induces a calmodulin-dependent alteration in phosphodiesterase activity of rat kidney in vivo.Endocrinology 112:1065–1069

    PubMed  Google Scholar 

  32. Misch, P., Giebel, P., Faust, R. 1982. Intestinal microvilli responses to feeding and fasting.Eur. J. Cell Biol. 21:264–279

    Google Scholar 

  33. Mooseker, M.S., Bonder, E.M., Conzelman, K.A., Fishkind, D.G., Howe, C.L., Keller, T.C.S. 1984. Brush border cytoskeleton and integration of cellular functions.J. Cell Biol. 99:104–112

    Google Scholar 

  34. Pearl, M., Taylor, A. 1983. High voltage electron microscopy (HVEM) of toad urinary bladder.J. Cell Biol. 97:312a (abstr).

    Google Scholar 

  35. Pollard, T. 1984. Effects of Ca2+ and Mg2− on actin polymerization.In: Calcium Regulation in Biological Systems. S. Ebashi, editor. pp. 71–81. Academic, Tokyo

    Google Scholar 

  36. Rasmussen, H., Goodman, D.B.P. 1977. Relationships between calcium and cyclic nucleotides in cell activation.Physiol. Rev. 57:421–509

    Google Scholar 

  37. Romey, G., Hugues, M., Schmid-Antomarchi, H., Lazdunski, M. 1984. Apamin: A specific toxin to study a class of Ca2+-dependent K+ channels.J. Physiol. (Paris) 79:259–264

    Google Scholar 

  38. Steiner, A.L., Pagliara, A.S., Chase, L.R., Kipnis, D.M. 1972. Radioimmunoassay for cyclic nucleotides. 1. Adenosine 3′5′-monophosphate and guanosine 3′5′-monophosphate in mammalian tissues and body fluids.J. Biol. Chem. 247:1114–1123

    PubMed  Google Scholar 

  39. Teitelbaum, A.P., Strewler, G.J. 1984. Parathyroid hormone receptors coupled to cyclic adenosine monophosphate formation in an established renal cell line.Endocrinology 114:980–985

    PubMed  Google Scholar 

  40. Tsien, R.H., Pozzan, T., Rink, T.J. 1982. Calcium homeostasis in intact lymphocytes: Cytoplasmic free calcium monitored with a new intracellularly trapped fluorescent indicator.J. Cell Biol. 94:325–334

    PubMed  Google Scholar 

  41. Vinay, P., Gougoux, A., Lemieux, G. 1981. Isolation of a pure suspension of rat proximal tubules.Am. J. Physiol. 241:F403-F411

    PubMed  Google Scholar 

  42. Warnock, D. 1984. Ion transport processes in apical membranes of epithelia. (Symposium Summary).Fed. Proc. 43:2473–2487

    Google Scholar 

  43. Weber, K., Glenney, J.R. 1982. Calcium-modulated multifunctional proteins regulating F-actin organization.Cold Spring Harbor Symp. Quant. Biol. 46:541–579

    PubMed  Google Scholar 

  44. Wulf, E., Deboben, A., Bautz, F.A., Faulstich, H., Wieland, T. 1979. Fluorescent phallotoxin, a tool for visualization of cellular actin.Proc. Natl. Acad. Sci. USA 76:4498–4502

    PubMed  Google Scholar 

  45. Yanagawa, N. 1985. Cytosolic calcium in isolated perfused rabbit renal proximal tubules: Heterogeneous response to PTH.Clin. Res. 33:503A (abstr.)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Goligorsky, M.S., Menton, D.N. & Hruska, K.A. Parathyroid hormone-induced changes of the brush border topography and cytoskeleton in cultured renal proximal tubular cells. J. Membrain Biol. 92, 151–162 (1986). https://doi.org/10.1007/BF01870704

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01870704

Key Words

Navigation