Logo of jbiocytThe Journal of Cell BiologyThis Article at jcb.orgEditorsContactThe Rockefeller University PressInstructions to Authors
J Biophys Biochem Cytol. Sep 25, 1957; 3(5): 705–724.
PMCID: PMC2224122

AN ANALYSIS OF MYOGENESIS BY THE USE OF FLUORESCENT ANTIMYOSIN

Abstract

Antibodies against myosin of adult chicken skeletal muscle were labelled with fluorescein and used as staining reagents to analyze the development of trunk myoblasts in the chick embryo. Myoblasts from the brachial myotomes were studied in three ways: (a) Specimens were fixed, sectioned, and stained with iron-hematoxylin. (b) Living myoblasts, and myoblasts prepared by glycerol extraction, were teased and examined by phase contrast microscopy. (c) Embryo trunks were treated with fluorescent antimyosin or with a control solution of fluorescent normal globulin, and were examined by fluorescence and phase contrast microscopy. Both glycerol-extracted and fixed materials were used. Cross-striated myofibrils appeared first in stage 16 to 17 embryos in the series studied by antimyosin staining and fluorescence microscopy. Striated myofibrils appeared first in stage 18 to 19 embryos, in the series stained by iron-hematoxylin, and at stage 22 to 23, in the series studied by glycerol extraction and phase contrast microscopy. In each series, myofibrils without apparent cross-striations were detected shortly before cross-striations were observed. Specific staining by antimyosin occurred only in differentiating myoblasts. Within the myoblasts antimyosin staining was confined to the A bands of the slender myofibrils. The following observations suggest that the first delicate striated structure to appear in the early 3 day myoblast was remarkably mature: (1) The sarcomere pattern both in length and in internal detail, was similar to that of adult muscle. (2) The distribution of myosin, as revealed by antimyosin staining, was the same in the embryonic as in the mature myofibril. (3) Glycerol-extracted myoblasts contracted vigorously on exposure to ATP. The changes in sarcomere band pattern were indistinguishable from those occurring during contraction of adult muscle induced by ATP. (4) ATP contraction was blocked by prior antimyosin staining in embryonic myoblasts as in mature muscle. It is suggested that the early myofibril grows laterally as a thin sheet associated with the sarcolemma, and that growth in length occurs in the growth tips of the elongating myoblast.

Full Text

The Full Text of this article is available as a PDF (1.3M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • ANFINSEN CB, REDFIELD RR. Protein structure in relation to function and biosynthesis. Adv Protein Chem. 1956;11:1–100. [PubMed]
  • ASHLEY CA, PORTER KR, PHILPOTT DE, HASS GM. Observations by electron microscopy on contraction of skeletal myofibrils induced with adenosinetriphosphate. J Exp Med. 1951 Jul 1;94(1):9–20. [PMC free article] [PubMed]
  • COONS AH, KAPLAN MH. Localization of antigen in tissue cells; improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med. 1950 Jan 1;91(1):1–13. [PMC free article] [PubMed]
  • CSAPO A, HERRMANN H. Quantitative changes in contractile proteins of chick skeletal muscle during and after embryonic development. Am J Physiol. 1951 Jun;165(3):701–710. [PubMed]
  • Ebert JD. An Analysis of the Synthesis and Distribution of the Contractile Protein, Myosin, in the Development of the Heart. Proc Natl Acad Sci U S A. 1953 Apr;39(4):333–344. [PMC free article] [PubMed]
  • FLEWETT TH, CHALLICE CE. The cutting of sections for electron microscopy using ester wax as an embedding medium. J Pathol Bacteriol. 1951 Oct;63(4):748–751. [PubMed]
  • FINCK H, HOLTZER H, MARSHALL JM., Jr An immunochemical study of the distribution of myosin in glycerol extracted muscle. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):175–178. [PMC free article] [PubMed]
  • HIBBS RG. Electron microscopy of developing cardiac muscle in chick embryos. Am J Anat. 1956 Jul;99(1):17–51. [PubMed]
  • HODGE AJ. Studies on the structure of muscle. III. Phase contrast and electron microscopy of dipteran plight muscle. J Biophys Biochem Cytol. 1955 Jul 25;1(4):361–380. [PMC free article] [PubMed]
  • HUGHES A. The growth of embryonic neurites; a study of cultures of chick neural tissues. J Anat. 1953 Apr;87(2):150–162. [PMC free article] [PubMed]
  • MOMMAERTS WFHM, PARRISH RG. Studies on myosin. I. Preparation and criteria of purity. J Biol Chem. 1951 Feb;188(2):545–552. [PubMed]
  • NEFF JC, BECKER EL. Antigen-antibody reactions in agar. III. Rate of change of band migration with antigen concentration. J Immunol. 1957 Jan;78(1):5–10. [PubMed]
  • SZENT-GYORGYI A. Free-energy relations and contraction of actomyosin. Biol Bull. 1949 Apr;96(2):140–161. [PubMed]
  • SZENT-GYORGYI AG. Meromyosins, the subunits of myosin. Arch Biochem Biophys. 1953 Feb;42(2):305–320. [PubMed]
  • SZENT-GYORGYI AG, MAZIA D, SZENT-GYORGYI A. On the nature of the cross-striation of body muscle. Biochim Biophys Acta. 1955 Mar;16(3):339–342. [PubMed]
  • WEBER HH, PORTZEHL H. Muscle contraction and fibrous muscle proteins. Adv Protein Chem. 1952;7:161–252. [PubMed]
  • WEISZ PB. Morphogenesis in Protozoa. Q Rev Biol. 1954 Sep;29(3):207–229. [PubMed]

Articles from The Journal of Biophysical and Biochemical Cytology are provided here courtesy of The Rockefeller University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • PubMed
    PubMed
    PubMed citations for these articles

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...