The growth and differentiation af aortal smooth muscle cells after calcitriol treatment are associated with microtubule reorganisation - an in vitro study

• Autorzy: Tukaj C , Bohdanowicz J. , Kubasik-Juraniec J.
• Języki publikacji: EN

Abstrakty

EN

The smooth muscle cells (SMCs) of the arterial media play a predominant role in functional and structural alterations of the arterial wall. The transition from the “contractile” to the “synthetic” phenotype appears to be an early critical event in the development of atherosclerotic disease. A number of observations suggest that 1,25(OH)₂D₃ (calcitriol) is of importance in maintaining normal cardiovascular function through its receptors in cardiac myocytes or aortal SMCs. The present study has focused on the microtubular (MT) network reorganisation after exposure to calcitriol. SMCs isolated by enzymatic digestion from the aortal media of neonatal rats were cultured on glass cover slips. 1 μM of 1,25(OH)₂D₃ was added to the culture medium every second day. The cytoskeletal features of SMCs after calcitriol were visualised by the immunofluorescence staining of α-tubulin. The alterations in α-tubulin expression and the distribution of microtubules related to the activities of the vascular smooth muscle cells, namely adhesion, migration, multilayer formation and cell division, were observed. A spindle shape, decreased cell adhesion, low expression of α-tubulin and a longitudinally arranged microtubular network manifested the high rate of SMC differentiation in the calcitriol-treated culture. A flat stellate morphology, high expression of α-tubulin and a radially distributed three-dimensional microtubular network were observed in the SMCs of the control culture. Destructive changes in the microtubular architecture which altered the cellular shape were evident in SMCs undergoing apoptosis. Cells with apoptotic features were more frequent in calcitriol-exposed culture. In contrast to the regular SMC divisions observed in the control culture, some of the mitotic cells exposed to calcitriol contained broader bipolar, multipolar or disordered spindles. These alterations in the SMCs’ microtubular cytoskeleton after calcitriol treatment were concomitant with changes in cell growth, differentiation and apoptosis, and may suggest a similarity to atherosclerotic plaque formation.

Słowa kluczowe

EN

cytoskeleton; tissue culture; reorganization; muscle cell; in vitro; growth; smooth muscle; differentiation

• Wydawca: -
• Czasopismo: Folia Morphologica
• Rocznik: 2004
• Tom: 63
• Numer: 1
• Opis fizyczny: p.51-57,fig.,ref.

Twórcy

autor

Tukaj C

• Medical University of Gdansk, Debinki 1, 80-211 Gdansk, Poland

autor

Bohdanowicz J.

autor

Kubasik-Juraniec J.

Bibliografia

• 1. Bochaton-Piallat ML, Ropraz P, Gabbiani F, Gabbiani G (1996) Phenotypic heterogeneity of rat arterial smooth muscle cell clones. Implications for the development of experimental intimal thickening. Arterioscler Thromb Vasc Biol, 16: 815–820.
• 2. Campbell JH, Kocher O, Skalli O, Gabbiani G, Campbell GR (1989) Cytodifferentiation and expression of alpha-smooth muscle actin mRNA and protein during primary culture of aortic smooth muscle cells: correlation with cell density and proliferative state. Arteriosclerosis, 9: 633–643.
• 3. Chamley-Campbell JH, Campbell GR, and Ross R (1979) The smooth muscle cell in culture. Physiol Rev, 59: 1–61.
• 4. Cassimeris L (1999) Accessory protein regulation of microtubule dynamics throughout the cell cycle. Curr Opin Cell Biol, 11: 134–141.
• 5. Chitaley K, Webb RC (2001) Microtubule depolymerization facilitates contraction of vascular smooth muscle via increased activation of RhoA/Rho-kinase. Med Hypotheses, 56: 381–385.
• 6. Heald R (2000) A dynamic duo of microtubule modulators. Nat Cell Biol, 2: E11–E12.
• 7. Hegele RA (1996) The pathogenesis of atherosclerosis. Clin Chim Acta, 146: 21–38.
• 8. Ireland CM, Pittman SM (1995) Tubulin alterations in taxol-induced apoptosis parallel those observed with other drugs. Biochem Pharmacol, 49: 1491–1499.
• 9. Jono S, Nishizawa Y, Shioi A, Morii H (1998) 1,25-Dihydroxyvitamin D3 increases in vitro vascular calcification by modulating secretion of endogenous parathyroid hormone-related peptide. Circulation, 98: 1302–1306.
• 10. Kockx MM, Knaapen MW (2000) The role of apoptosis in vascular disease. J Pathol, 190: 267–280.
• 11. Koh E, Morimoto S, Fukuo K, Hironaka T, Onishi T, Kumahara Y (1988) 1,25-dihydroxyvitamin D3 binds specifically to rat vascular smooth muscle cells and stimulate their proliferation in vitro. Life Sci, 42: 315–323.
• 12. Lang F, Ritter M, Gamper N, Huber S, Fillon S, Tanneur V, Lepple-Wienhues A, Szabo I, Gulbins E (2000) Cell volume in the regulation of cell proliferation and apoptotic cell death. Cell Physiol Biochem, 10: 417–428.
• 13. Li Z, Cheng H, Lederer WJ, Froehlich J, Lakatta EG (1997) Enhanced proliferation and migration and altered cytoskeletal proteins in early passage smooth muscle cells from young and old rat aortic explants. Exp Mol Pathol, 64: 1–11.
• 14. Mayr M, Xu Q (2001) Smooth muscle cell apoptosis in arteriosclerosis. Exp Gerontol, 36: 968–687.
• 15. Merke J, Hofmann W, Goldschmidt D, Ritz E (1987) Demonstration of 1,25(OH)2 vitamin D3 receptors and actions in vascular smooth muscle cells in vitro. Calcif Tissue Int, 41: 112–114.
• 16. Mitsuhashi T, Morris RC, Ives HE (1991) 1,25-dihydroxyvitamin D₃ modulates growth of vascular smooth muscle cells. J Clin Invest, 87: 1889–1895.
• 17. Nabi IR (1999) The polarization of the motile cell. J Cell Sci, 112: 1803–1811.
• 18. Owens GK (1995) Regulation of differentiation of vascular smooth muscle cells. Physiol Rev, 75: 487–517.
• 19. Reichel H, Koeffler HP, Norman AW (1989) The role of the vitamin D endocrine system in health and disease. New Engl J Med, 320: 980–981.
• 20. Sato N, Mizumoto K, Nakamura M, Maehara N, Minamishima Y, Nishio S, Nagai E, Tanaka M (2001) Correlation between centrosome abnormalities and chromosomal instability in human pancreatic cancer cells. Cancer Gen Cytogen, 126: 13–19.
• 21. Skalli O, Bloom WS, Ropraz P, Azzarone B, Gabbiani G (1986) Cytoskeletal remodeling of rat aortic smooth muscle cells in vitro: relationships to culture conditions and analogies to in vivo situations. J Submicrosc Cytol, 18: 481–493.
• 22. Tao F, Chaudry S, Tolloczko B, Martin JG, Kelly SM (2003) Modulation of smooth muscle phenotype in vitro by homologous cell substrate. Am J Physiol Cell Physiol, 284: C1531–C154.
• 23. Thoumine O, Ott A (1996) Influence of adhesion and cytoskeletal integrity on fibroblast traction. Cell Motil Cytoskeleton, 35: 269–280.
• 24. Thyberg J, Hedin U, Sjölund M, Palmberg L, Bottger BA (1990) Regulation of differentiated properties and proliferation of arterial smooth muscle cells. Arteriosclerosis, 10: 966–990.
• 25. Tukaj C, Kubasik-Juraniec J, Kraszpulski M (2000) Morphological changes of aortal smooth muscle cells exposed to calcitriol in culture. Med Sci Monit, 6: 668–674.
• 26. Tukaj C, Bohdanowicz J, Kubasik-Juraniec J (2000) Effects of 1,25-dihydroxycholecalciferol on cytoskeleton organization of aortal smooth muscle cells in primary culture. In: Norman AW, Bouillon R, Thomasset M (eds.). Vitamin D Endocrine System: structural, biological, genetic and clinical aspects. Univ. California, Riverside, pp. 431–434.
• 27. Tukaj C, Bohdanowicz J, Kubasik-Juraniec J (2002) A scanning electron microscopic study of phenotypic plasticity and surface structural changes of aortal smooth muscle cells in primary culture. Folia Morphol, 61: 191–198.
• 28. Worth NF, Rolfe BE, Song J, Campbell GR (2001) Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteins. Cell Motil Cytoskeleton, 49: 130–145.