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. 1980 Dec 1;87(3):594–600. doi: 10.1083/jcb.87.3.594

Permanently proliferating rat vascular smooth muscle cell with maintained expression of smooth muscle characteristics, including actin of the vascular smooth muscle type

PMCID: PMC2110800  PMID: 6893986

Abstract

Cells of an established clonal line (RVF-SMC) derived from rat vena cava are described by light and electron microscope methods and biochemical analysis of the major proteins. The cells are flat, and they moderately elongate and form monolayers. They are characterized by prominent cables of microfilaments bundles decoratable with antibodies to actin and alpha-actinin. These bundles contain numerous densely stained bodies and are often flanked by typical rows of surface caveolae and vesicles. The cells are rich in intermediate-sized filaments of the vimentin type but do not show detectable amounts of desmin and cytokeratin filaments. Isoelectric focusing and protein chemical studies have revealed actin heterogeneity. In addition to the two cytoplasmic actins, beta and gamma, common to proliferating cells, two smooth muscle-type actins (an acidic alpha-like and a gamma-like) are found. The major (alpha-type) vascular smooth muscle actin accounts for 28% of the total cellular actin. No skeletal muscle or cardiac muscle actin has been detected. The synthesis of large amounts of actin and vimentin and the presence of at least three actins, including alpha- like actin, have also been demonstrated by in vitro translation of isolated poly(A)+ mRNAs. This is, to our knowledge, the first case of expression of smooth muscle-type actin in a permanently growing cell. We conclude that permanent cell growth and proliferation is compatible with the maintained expression of several characteristic cell features of the differentiated vascular smooth muscle cell including the formation of smooth muscle-type actin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett G. S., Fellini S. A., Croop J. M., Otto J. J., Bryan J., Holtzer H. Differences among 100-A filamentilament subunits from different cell types. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4364–4368. doi: 10.1073/pnas.75.9.4364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burke J. M., Ross R. Collagen synthesis by monkey arterial smooth muscle cells during proliferation and quiescence in culture. Exp Cell Res. 1977 Jul;107(2):387–395. doi: 10.1016/0014-4827(77)90360-3. [DOI] [PubMed] [Google Scholar]
  4. Burridge K. A comparison of fibroblast and smooth muscle myosins. FEBS Lett. 1974 Sep 1;45(1):14–17. doi: 10.1016/0014-5793(74)80799-4. [DOI] [PubMed] [Google Scholar]
  5. Campbell G. R., Uehara Y., Mark G., Burnstock G. Fine structure of smooth muscle cells grown in tissue culture. J Cell Biol. 1971 Apr;49(1):21–34. doi: 10.1083/jcb.49.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chamley-Campbell J., Campbell G. R., Ross R. The smooth muscle cell in culture. Physiol Rev. 1979 Jan;59(1):1–61. doi: 10.1152/physrev.1979.59.1.1. [DOI] [PubMed] [Google Scholar]
  7. Chamley J. H., Gröschel-Stewart U., Campbell G. R., Burnstock G. Distinction between smooth muscle, fibroblasts and endothelial cells in culture by the use of fluoresceinated antibodies against smooth muscle actin. Cell Tissue Res. 1977 Feb 14;177(4):445–457. doi: 10.1007/BF00220606. [DOI] [PubMed] [Google Scholar]
  8. Fowler S., Shio H., Wolinsky H. Subcellular fractionation and morphology of calf aortic smooth muscle cells. Studies on whole aorta, aortic explants, and subcultures grown under different conditions. J Cell Biol. 1977 Oct;75(1):166–184. doi: 10.1083/jcb.75.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Franke W. W., Schmid E., Freudenstein C., Appelhans B., Osborn M., Weber K., Keenan T. W. Intermediate-sized filaments of the prekeratin type in myoepithelial cells. J Cell Biol. 1980 Mar;84(3):633–654. doi: 10.1083/jcb.84.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Franke W. W., Schmid E., Osborn M., Weber K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5034–5038. doi: 10.1073/pnas.75.10.5034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Franke W. W., Schmid E., Winter S., Osborn M., Weber K. Widespread occurrence of intermediate-sized filaments of the vimentin-type in cultured cells from diverse vertebrates. Exp Cell Res. 1979 Oct 1;123(1):25–46. doi: 10.1016/0014-4827(79)90418-x. [DOI] [PubMed] [Google Scholar]
  12. Fyrberg E. A., Donady J. J. Actin Heterogeneity in primary embryonic culture cells from Drosophila melanogaster. Dev Biol. 1979 Feb;68(2):487–502. doi: 10.1016/0012-1606(79)90220-3. [DOI] [PubMed] [Google Scholar]
  13. Garrels J. I., Gibson W. Identification and characterization of multiple forms of actin. Cell. 1976 Dec;9(4 Pt 2):793–805. doi: 10.1016/0092-8674(76)90142-2. [DOI] [PubMed] [Google Scholar]
  14. Gimbrone M. A., Jr, Cotran R. S. Human vascular smooth muscle in culture. Growth and ultrastructure. Lab Invest. 1975 Jul;33(1):16–27. [PubMed] [Google Scholar]
  15. Gruenstein E., Rich A. Non-identity of muscle and non-muscle actins. Biochem Biophys Res Commun. 1975 May 19;64(2):472–477. doi: 10.1016/0006-291x(75)90345-9. [DOI] [PubMed] [Google Scholar]
  16. Horovitch S. J., Storti R. V., Rich A., Pardue M. L. Multiple actins in Drosophila melanogaster. J Cell Biol. 1979 Jul;82(1):86–92. doi: 10.1083/jcb.82.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hunter T., Garrels J. I. Characterization of the mRNAs for alpha-, beta- and gamma-actin. Cell. 1977 Nov;12(3):767–781. doi: 10.1016/0092-8674(77)90276-8. [DOI] [PubMed] [Google Scholar]
  18. Izant J. G., Lazarides E. Invariance and heterogeneity in the major structural and regulatory proteins of chick muscle cells revealed by two-dimensional gel electrophoresis. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1450–1454. doi: 10.1073/pnas.74.4.1450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kelly P. T., Cotman C. W. Synaptic proteins. Characterization of tubulin and actin and identification of a distinct postsynaptic density polypeptide. J Cell Biol. 1978 Oct;79(1):173–183. doi: 10.1083/jcb.79.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kimes B. W., Brandt B. L. Characterization of two putative smooth muscle cell lines from rat thoracic aorta. Exp Cell Res. 1976 Mar 15;98(2):349–366. doi: 10.1016/0014-4827(76)90446-8. [DOI] [PubMed] [Google Scholar]
  21. Lazarides E., Hubbard B. D. Immunological characterization of the subunit of the 100 A filaments from muscle cells. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4344–4348. doi: 10.1073/pnas.73.12.4344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lazarides E., Lindberg U. Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4742–4746. doi: 10.1073/pnas.71.12.4742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mayne R., Vail M. S., Mayne P. M., Miller E. J. Changes in type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity. Proc Natl Acad Sci U S A. 1976 May;73(5):1674–1678. doi: 10.1073/pnas.73.5.1674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  25. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rubenstein P. A., Spudich J. A. Actin microheterogeneity in chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Jan;74(1):120–123. doi: 10.1073/pnas.74.1.120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Storti R. V., Coen D. M., Rich A. Tissue-specific forms of actin in the developing chick. Cell. 1976 Aug;8(4):521–527. doi: 10.1016/0092-8674(76)90220-8. [DOI] [PubMed] [Google Scholar]
  28. Storti R. V., Rich A. Chick cytoplasmic actin and muscle actin have different structural genes. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2346–2350. doi: 10.1073/pnas.73.7.2346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vandekerckhove J., Weber K. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 1978 Dec 25;126(4):783–802. doi: 10.1016/0022-2836(78)90020-7. [DOI] [PubMed] [Google Scholar]
  30. Vandekerckhove J., Weber K. Mammalian cytoplasmic actins are the products of at least two genes and differ in primary structure in at least 25 identified positions from skeletal muscle actins. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1106–1110. doi: 10.1073/pnas.75.3.1106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vandekerckhove J., Weber K. The complete amino acid sequence of actins from bovine aorta, bovine heart, bovine fast skeletal muscle, and rabbit slow skeletal muscle. A protein-chemical analysis of muscle actin differentiation. Differentiation. 1979;14(3):123–133. doi: 10.1111/j.1432-0436.1979.tb01021.x. [DOI] [PubMed] [Google Scholar]

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