PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
 
J Cell Biol. 1986 December 1; 103(6): 2787–2796.
PMCID: PMC2114627

A monoclonal antibody against alpha-smooth muscle actin: a new probe for smooth muscle differentiation

Abstract

A monoclonal antibody (anti-alpha sm-1) recognizing exclusively alpha- smooth muscle actin was selected and characterized after immunization of BALB/c mice with the NH2-terminal synthetic decapeptide of alpha- smooth muscle actin coupled to keyhole limpet hemocyanin. Anti-alpha sm- 1 helped in distinguishing smooth muscle cells from fibroblasts in mixed cultures such as rat dermal fibroblasts and chicken embryo fibroblasts. In the aortic media, it recognized a hitherto unknown population of cells negative for alpha-smooth muscle actin and for desmin. In 5-d-old rats, this population is about half of the medial cells and becomes only 8 +/- 5% in 6-wk-old animals. In cultures of rat aortic media SMCs, there is a progressive increase of this cell population together with a progressive decrease in the number of alpha- smooth muscle actin-containing stress fibers per cell. Double immunofluorescent studies carried out with anti-alpha sm-1 and anti- desmin antibodies in several organs revealed a heterogeneity of stromal cells. Desmin-negative, alpha-smooth muscle actin-positive cells were found in the rat intestinal muscularis mucosae and in the dermis around hair follicles. Moreover, desmin-positive, alpha-smooth muscle actin- negative cells were identified in the intestinal submucosa, rat testis interstitium, and uterine stroma. alpha-Smooth muscle actin was also found in myoepithelial cells of mammary and salivary glands, which are known to express cytokeratins. Finally, alpha-smooth muscle actin is present in stromal cells of mammary carcinomas, previously considered fibroblastic in nature. Thus, anti-alpha sm-1 antibody appears to be a powerful probe in the study of smooth muscle differentiation in normal and pathological conditions.

Full Text

The Full Text of this article is available as a PDF (3.6M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barja F, Coughlin C, Belin D, Gabbiani G. Actin isoform synthesis and mRNA levels in quiescent and proliferating rat aortic smooth muscle cells in vivo and in vitro. Lab Invest. 1986 Aug;55(2):226–233. [PubMed]
  • Benyamin Y, Roustan C, Boyer M. Anti-actin antibodies. Chemical modification allows the selective production of antibodies to the N-terminal region. J Immunol Methods. 1986 Jan 22;86(1):21–29. [PubMed]
  • Bulinski JC, Kumar S, Titani K, Hauschka SD. Peptide antibody specific for the amino terminus of skeletal muscle alpha-actin. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1506–1510. [PubMed]
  • Chaponnier C, Patebex P, Gabbiani G. Human plasma actin-depolymerizing factor. Purification, biological activity and localization in leukocytes and platelets. Eur J Biochem. 1985 Jan 15;146(2):267–276. [PubMed]
  • Franke WW, Schmid E, Freudenstein C, Appelhans B, Osborn M, Weber K, Keenan TW. Intermediate-sized filaments of the prekeratin type in myoepithelial cells. J Cell Biol. 1980 Mar;84(3):633–654. [PMC free article] [PubMed]
  • Franke WW, Schmid E, Vandekerckhove J, Weber K. Permanently proliferating rat vascular smooth muscle cell with maintained expression of smooth muscle characteristics, including actin of the vascular smooth muscle type. J Cell Biol. 1980 Dec;87(3 Pt 1):594–600. [PMC free article] [PubMed]
  • Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR, Majno G. Granulation tissue as a contractile organ. A study of structure and function. J Exp Med. 1972 Apr 1;135(4):719–734. [PMC free article] [PubMed]
  • Gabbiani G, Kocher O, Bloom WS, Vandekerckhove J, Weber K. Actin expression in smooth muscle cells of rat aortic intimal thickening, human atheromatous plaque, and cultured rat aortic media. J Clin Invest. 1984 Jan;73(1):148–152. [PMC free article] [PubMed]
  • Gabbiani G, Schmid E, Winter S, Chaponnier C, de Ckhastonay C, Vandekerckhove J, Weber K, Franke WW. Vascular smooth muscle cells differ from other smooth muscle cells: predominance of vimentin filaments and a specific alpha-type actin. Proc Natl Acad Sci U S A. 1981 Jan;78(1):298–302. [PubMed]
  • Garrels JI, Gibson W. Identification and characterization of multiple forms of actin. Cell. 1976 Dec;9(4 Pt 2):793–805. [PubMed]
  • Gleiber WE, Schiffmann E. Identification of a chemoattractant for fibroblasts produced by human breast carcinoma cell lines. Cancer Res. 1984 Aug;44(8):3398–3402. [PubMed]
  • Gown AM, Vogel AM, Gordon D, Lu PL. A smooth muscle-specific monoclonal antibody recognizes smooth muscle actin isozymes. J Cell Biol. 1985 Mar;100(3):807–813. [PMC free article] [PubMed]
  • Herman IM, D'Amore PA. Microvascular pericytes contain muscle and nonmuscle actins. J Cell Biol. 1985 Jul;101(1):43–52. [PMC free article] [PubMed]
  • Hori K, Hashimoto K, Eto H, Dekio S. Keratin type intermediate filaments in sweat gland myoepithelial cells. J Invest Dermatol. 1985 Nov;85(5):453–459. [PubMed]
  • Ives HE, Schultz GS, Galardy RE, Jamieson JD. Preparation of functional smooth muscle cells from the rabbit aorta. J Exp Med. 1978 Nov 1;148(5):1400–1413. [PMC free article] [PubMed]
  • Keller O, Rudinger J. Preparation and some properties of maleimido acids and maleoyl derivatives of peptides. Helv Chim Acta. 1975 Mar 12;58(2):531–541. [PubMed]
  • Kocher O, Gabbiani G. Cytoskeletal features of normal and atheromatous human arterial smooth muscle cells. Hum Pathol. 1986 Sep;17(9):875–880. [PubMed]
  • Kocher O, Skalli O, Bloom WS, Gabbiani G. Cytoskeleton of rat aortic smooth muscle cells. Normal conditions and experimental intimal thickening. Lab Invest. 1984 Jun;50(6):645–652. [PubMed]
  • Kocher O, Skalli O, Cerutti D, Gabbiani F, Gabbiani G. Cytoskeletal features of rat aortic cells during development. An electron microscopic, immunohistochemical, and biochemical study. Circ Res. 1985 Jun;56(6):829–838. [PubMed]
  • Kuroda M. Change of actin isomers during differentiation of smooth muscle. Biochim Biophys Acta. 1985 Dec 13;843(3):208–213. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Larson DM, Fujiwara K, Alexander RW, Gimbrone MA., Jr Heterogeneity of myosin antigenic expression in vascular smooth muscle in vivo. Lab Invest. 1984 Apr;50(4):401–407. [PubMed]
  • Leavitt J, Gunning P, Kedes L, Jariwalla R. Smooth muscle alpha-action is a transformation-sensitive marker for mouse NIH 3T3 and Rat-2 cells. Nature. 316(6031):840–842. [PubMed]
  • Leavitt J, Leavitt A, Attallah AM. Dissimilar modes of expression of beta- and gamma-actin in normal and leukemic human T lymphocytes. J Biol Chem. 1980 Jun 10;255(11):4984–4987. [PubMed]
  • Longtine JA, Pinkus GS, Fujiwara K, Corson JM. Immunohistochemical localization of smooth muscle myosin in normal human tissues. J Histochem Cytochem. 1985 Mar;33(3):179–184. [PubMed]
  • Lubit BW, Schwartz JH. Immunological characterization of an anti-actin antibody specific for cytoplasmic actins and its use for the immunocytological localization of actin in Aplysia nervous tissue. J Histochem Cytochem. 1983 Jun;31(6):728–736. [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed]
  • Osborn M, Weber K. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest. 1983 Apr;48(4):372–394. [PubMed]
  • Otey CA, Kalnoski MH, Lessard JL, Bulinski JC. Immunolocalization of the gamma isoform of nonmuscle actin in cultured cells. J Cell Biol. 1986 May;102(5):1726–1737. [PMC free article] [PubMed]
  • Owens GK, Loeb A, Gordon D, Thompson MM. Expression of smooth muscle-specific alpha-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation. J Cell Biol. 1986 Feb;102(2):343–352. [PMC free article] [PubMed]
  • Pardo JV, Pittenger MF, Craig SW. Subcellular sorting of isoactins: selective association of gamma actin with skeletal muscle mitochondria. Cell. 1983 Apr;32(4):1093–1103. [PubMed]
  • Rosenberg S, Stracher A, Lucas RC. Isolation and characterization of actin and actin-binding protein from human platelets. J Cell Biol. 1981 Oct;91(1):201–211. [PMC free article] [PubMed]
  • Roustan C, Benyamin Y, Boyer M, Cavadore JC. Structural variations in actins. A study of the immunological reactivity of the N-terminal region. Biochem J. 1986 Jan 1;233(1):193–197. [PubMed]
  • Rubenstein PA. Differential behavior of gizzard isoactins. Arch Biochem Biophys. 1981 Sep;210(2):598–608. [PubMed]
  • Rungger-Brändle E, Gabbiani G. The role of cytoskeletal and cytocontractile elements in pathologic processes. Am J Pathol. 1983 Mar;110(3):361–392. [PubMed]
  • Saurat JH, Didierjean L, Skalli O, Siegenthaler G, Gabbiani G. The intermediate filament proteins of rabbit normal epidermal Merkel cells are cytokeratins. J Invest Dermatol. 1984 Dec;83(6):431–435. [PubMed]
  • Schmid E, Osborn M, Rungger-Brändle E, Gabbiani G, Weber K, Franke WW. Distribution of vimentin and desmin filaments in smooth muscle tissue of mammalian and avian aorta. Exp Cell Res. 1982 Feb;137(2):329–340. [PubMed]
  • Schürch W, Lagacé R, Seemayer TA. Myofibroblastic stromal reaction in retracted scirrhous carcinoma of the breast. Surg Gynecol Obstet. 1982 Mar;154(3):351–358. [PubMed]
  • Simpson PA, Spudich JA, Parham P. Monoclonal antibodies prepared against Dictyostelium actin: characterization and interactions with actin. J Cell Biol. 1984 Jul;99(1 Pt 1):287–295. [PMC free article] [PubMed]
  • Skalli O, Bloom WS, Ropraz P, Azzarone B, Gabbiani G. Cytoskeletal remodeling of rat aortic smooth muscle cells in vitro: relationships to culture conditions and analogies to in vivo situations. J Submicrosc Cytol. 1986 Jul;18(3):481–493. [PubMed]
  • Storti RV, 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. [PubMed]
  • Strauch AR, Rubenstein PA. Induction of vascular smooth muscle alpha-isoactin expression in BC3H1 cells. J Biol Chem. 1984 Mar 10;259(5):3152–3159. [PubMed]
  • Takeuchi K. Differences between smooth and skeletal muscle myosins in their interactions with F-actin. J Biochem. 1982 Mar;91(3):1001–1007. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Travo P, Weber K, Osborn M. Co-existence of vimentin and desmin type intermediate filaments in a subpopulation of adult rat vascular smooth muscle cells growing in primary culture. Exp Cell Res. 1982 May;139(1):87–94. [PubMed]
  • Tremblay G. Stromal aspects of breast carcinoma. Exp Mol Pathol. 1979 Aug;31(1):248–260. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • Vandekerckhove J, Weber K. Actin typing on total cellular extracts: a highly sensitive protein-chemical procedure able to distinguish different actins. Eur J Biochem. 1981 Jan;113(3):595–603. [PubMed]
  • Wehland J, Schröder HC, Weber K. Amino acid sequence requirements in the epitope recognized by the alpha-tubulin-specific rat monoclonal antibody YL 1/2. EMBO J. 1984 Jun;3(6):1295–1300. [PubMed]
  • Whalen RG, Butler-Browne GS, Gros F. Protein synthesis and actin heterogeneity in calf muscle cells in culture. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2018–2022. [PubMed]
  • Witt DP, Brown DJ, Gordon JA. Transformation-sensitive isoactin in passaged chick embryo fibroblasts transformed by Rous sarcoma virus. J Cell Biol. 1983 Jun;96(6):1766–1771. [PMC free article] [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press