Skip to main content
Log in

Immunocytochemistry of the microtubules of fat-laden cells

Brown fat cells and adrenocortical cells in primary monolayer culture

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

Cytoplasmic microtubules of fat-laden cells containing fine lipid droplets, such as brown fat cells and adrenocortical cells, were studied in relation to the metabolism of intracellular lipid. In these cells, the amount and distribution of lipid droplets reflect the state of inherent cellular function. Materials used were primary monolayer culture of fetal rat brown fat cells and that of bovine adrenocortical cells. The method was the immunocytochemistry with anti-tubulin antibody. When brown fat cells were being lipolyzed or the steroidogenesis of adrenocortical cells were being stimulated, the cytoplasmic microtubules in the cells were organized in a radial pattern in response to the behavior of the lipid droplets. It is assumed that the microtubules were in the regulation of cellular function in terms of the metabolism of lipid droplets in these cells. We have devised, in the course of the current study, a double fluorescence technique as an observational method whereby microtubules were observed immunocytochemically and lipid droplets by a secondary fluorescence with phosphine E staining.

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

  • Adler KB, Brody AR, Craighead JE (1981) Studies on the mechanism of mucin secretion of cells of the porcine tracheal epithelium. Proc Soc Exp Biol Med 166:96–106

    Google Scholar 

  • Bickle D, Tilney LG, Porter KR (1966) Microtubules and pigment migration in the melanophores of fundulus heteroclitus L. Protoplasma 61:322–345

    Google Scholar 

  • Bohn W (1978) A fixation method for improved antibody penetration in electron microscopal immunoperoxidase studies. J Histochem Cytochem 26:293–297

    Google Scholar 

  • Byers HR, Fujiwara K, Beckerle MC, Porter KR (1980) Microtubule organization and function in chromatophores and scleroblasts in vitro and in situ. In: De Brabander M, De Mey J (eds) Microtubule and microtubule inhibitor. Elsevier/North-Holland, Amsterdam, pp 43–56

    Google Scholar 

  • Clark MA, Shay JW (1981) The role of tubulin in the steroidogenic response of murine adrenal and rat leydig cells. Endocrinology 109:2261–2263

    Google Scholar 

  • Dales S, Hsu KC, Nagayama A (1973) The fine structure and immunological labelling of the achromatic mitotic apparatus after disruption of cell membrane. J Cell Biol 59:643–660

    Google Scholar 

  • Eggstein M, Kuhlman E (1974) Glycerol. In: Bergmeyer HU (ed) Method of enzymic analysis, vol 4. Verlag Chemie, Weinheim/ Academic Press, New York and London, pp 1825–1831

    Google Scholar 

  • Hornsby PJ, Gill GN (1978) Characterization of adult bovine adrenocortical cells throughout their life span in tissue culture. Endocrinology 102:926–936

    Google Scholar 

  • Le Marchand Y, Singh A, Patzelt C, Jeanrenaud B (1975) In vivo and in vitro evidences for a role of microtubules in the secretory processes of liver. In: Borgers M, De Brabander M (eds) Microtubules and microtubules inhibitor. Elsevier/North-Holland, Amsterdam, pp 153–164

    Google Scholar 

  • Levy H, Deane HW, Rubin BL (1959) Visualization of steroid-3β-ol-dehydrogenase activity in tissue of intact and hypophysectomized rats. Endocrinology 65:932–943

    Google Scholar 

  • Malaisse WJ, Leclercq-Meyer V, Van Obberghen E, Somers G, Devis G, Ravazzola M, Malaisse-Lagae F, Orci L (1975) The role of the microtubular-microfilamentous system in insulin and glucagon release by the endocrine pancreas. In: Borgers M, De Brabander M (eds) Microtubules and microtubules inhibitor. Elsevier/North-Holland, Amsterdam, pp 143–152

    Google Scholar 

  • Murphy DB, Tilney LG (1974) The role of microtubules in the movement of pigment granules in toleost melanophores. J Cell Biol 61:757–779

    Google Scholar 

  • Nagayama A, Dales S (1970) Rapid purification and the immunological specificity of mammalian microtubular paracrystals possessing an ATPase activity. Proc Natl Acad Sci USA 66:464–471

    Google Scholar 

  • Neve P, Ketelbant-Balasse P, Willems C, Dumont JE (1972) Effect of inhibitors of microtubules and microfilaments on dog thyroid slices in vitro. Exp Cell Res 74:227–244

    Google Scholar 

  • Osborn M, Weberk (1976) Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane. Proc Natl Acad Sci USA 73:867–871

    Google Scholar 

  • Pretki M, Crettaz M, Jeanrenaud B (1981) Role of microtubules in insulin and glucagon stimulation of amino acid transport in isolated rat hepatocytes. J Biol Chem 256:4336–4340

    Google Scholar 

  • Schliwa M, (1975) Microtubule distribution and melanosome movements in fish melanophores. In: Borgers M, De Brabander M (eds) Microtubules and microtubule inhibitor. Elsevier/North-Holland, Amsterdam, pp 215–228

    Google Scholar 

  • Schliwa M, Osborn M, Weber K (1978) Microtubules system of isolated fish melanophores as revealed by immunofluorescence microscopy. J Cell Biol 76:229–236

    Google Scholar 

  • Silverstein SC, Steinman RM, Cohn ZA (1977) Endocytosis. Annu Rev Biochem 46:669–722

    Google Scholar 

  • Slautterback DB (1961) A fine tubular component of secretory cells. Am Soc Cell Biol (Abstr) 199, cf (82)

  • Sugihara H, Miyahara S, Ohta K, Yonemitsu N (1983) Hormonal sensitivity of brown fat cells of fetal rats in monolayer culture. Exp Clin Endocrinol (in press)

  • Valk BW, Popper H (1944) Microscopic demonstration of rat in urine and stool by means of fluorescence microscopy. Am J Clin Pathol 14:234

    Google Scholar 

  • Weber K, Pollack R, Bibring T (1975) Antibody against tubulin: The specific visualization of cytoplasmic microtubules in tissue culture cells. Proc Natl Acad Sci USA 72:459–463

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sugihara, H., Yonemitsu, N., Ohta, K. et al. Immunocytochemistry of the microtubules of fat-laden cells. Histochemistry 79, 227–236 (1983). https://doi.org/10.1007/BF00489784

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00489784

Keywords

Navigation