Summary
In cell cultures of skeletal muscles from 11 day-old chick embryos, myoblasts begin to fuse to myotubes on the second day of culture. Morphological data such as area of projection of the nucleus, length of myotubes and number of nuclei per myotube, as well as histochemical data such as relative DNA content and degree of chromatin condensation were evaluated. The myotubes examined, up to 216 h in culture, had an average of 7 nuclei. These nuclei, which had ceased DNA synthesis prior to cell fusion had less dense chromatin than the nuclei of myoblasts. The data which represent DNA condensation support the concept that transcriptional activity is enhanced during de novo synthesis of muscle protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Al J, Ploem JS (1979) Detection of suspicious cells and rejection of artefacts in cervical cytology using the Leyden television analysis system. J Histochem Cytochem 27:629–634
Anderson MJ, Cohen MW, Zorychta E (1977) Effects of innervation on the distribution of acetylcholine receptors on cultured muscle cells. J Physiol 268:731–756
Bachmann P, Hinrichsen K (1979) Principles and methods for the quantitative determination of Feulgen stained DNA with the television texture analysis system (TAS). Histochemistry 60:61–69
Bachmann P (1980) Motility, linear arrangement and cell-to-cell contact of myogenic cells prior to fusion. Cell Tissue Res 206:431–440
Betz FH, Firket H, Reznik M (1966) Some aspects of muscle regeneration. Int Rev Cytol 19:203–227
Bischoff R, Holtzer H (1969) Mitosis and the processes of differentiation of myogenic cells in vitro. J Cell Biol 41:188–200
Bishop RP, Young IT (1977) The automated classification of mitotic phases for human chromosome spreads. J Histochem Cytochem 25:730–740
Buckingham ME (1977) Muscle protein synthesis and its control during the differentiation of skeletal muscle cells in vitro. Int Rev Biochem 15:269–332
Capers CR (1960) Multinucleation of skeltal muscle in vitro. J Biophys Biochem Cytol 7:559–565 (1960)
Decher J, Bachmann P (1980) The influence of preset integer intervals at the densitometer unit of the television texture analysis system (TAS) on DNA quantification. Histochemistry 65:309–314
Duerksen JD, McCarthy BJ (1971) Distribution of deoxyribonucleic acid sequences in fractionated chromatin. Biochemistry 10:1471–1478
Friedlander M, Beyer E.C, Fischman DA (1978) Muscle development in vitro following X irradiation. Dev Biol 66:457–469
Graumann W (1953) Zur Standardisierung des Schiffschen Reagenz. Z Wiss Mikrosk 61:225–226 (1953)
Kalderon N, Epstein ML, Gilula NB (1977) Cell-to-cell communication and myogenesis. J Cell Biol 75:788–806
Lewis MR (1915) Rhythmical contraction of the skeletal muscle tissue observed in tissue cultures. Am J Physiol 38:153–161
Lewis WH, Lewis MR (1917) Behaviour of cross striated muscle in tissue cultures. Am J Anat 22:169–194
Littau VC, Allfrey VG, Frenster JH, Mirsky AE (1964) Active and inactive regions of nuclear chromatin as revealed by electron microscope autoradiography. Proc Natl Acad Sci USA 52:93–100
Merz M, Hinrichsen K (1969a) Erfassung morphologischer Merkmale von Zellkernen mittels bildanalytischer Meßdaten. I. Versuche zur densitometrischen Eichbarkeit des Quantimet. Prakt Metallogr 6:676–688
Merz M, Hinrichsen K (1969b) Erfassung morphologischer Merkmale von Zellkernen mittels bildanalytischer Meßdaten. II. Die Flächenrepräsentanz der Absorption. Prakt Metallogr 6:711–722
Meyer W, Hinrichsen K (1976) Veränderungen der Kernstruktur durch Actinomycin und ihre Erfassung mit Hilfe der Quantitativen Bildanalyse. Verh Anat Ges 70:923–926
Murphy EC, Hall SH, Shepherd JH, Weiser RS (1973) Fractionation of mouse myeloma chromatin. Biochemistry 12:3843–3853
Okazaki K, Holtzer H (1966) Myogenesis: fusion, myosin synthesis, and mitotic cycle. Proc Natl Acad Sci USA 56:1484–1490
Paterson B, Strohmann RC (1972) Myosin synthesis in cultures of differentiating chicken embryo skeletal muscle. Dev Biol 29:113–138
Ploem JS, Verwoerd N, Bonnet J, Koper G (1979) An automated microscope for quantitative cytology combining television image analysis and stage scanning microphotometry. J Histochem Cytochem 27:136–143
Rodriguez LV, Becker FF (1976) Rat liver chromatin. Fractionation into eu- and heterochromatin with localization of ribosomal genes. Arch Biochem Biophys 173:428–437
Shainberg A, Yagil G, Yaffe D (1971) Alterations of enzymatic activities during muscle differentiation in vitro. Dev Biol 25:1–29
Slater CR (1976) Control of myogenesis in vitro by chick embryo extract. Dev Biol 50:264–284
Stockdale FE, Holtzer H (1961) DNA synthesis and myogenesis. Exp Cell Res 24:508–520
Strehler BL, Konigsberg IR, Kelly JET (1963) Ploidy of myotube nuclei developing in vitro as determined with a recording double beam microspectrophotometer. Exp Cell Res 32:232–241
Tanke HJ, Ingen EM van, Pleom JS (1979) Acriflavine-Feulgen stilbene (AFS) staining: a procedure for automated cervical cytology with a television based system (LEYTAS) J Histochem Cytochem 27:84–86
Tata JR, Baker B (1974) Sub-nuclear fractionation. II. Intranuclear compartmentation of transcription in vivo and in vitro. Exp Cell Res 83:125–138
Turner DC, Maier V, Eppenberger, HM (1974) Creatine kinase and aldolase isoenzyme transitions in cultures of chick skeletal muscle cells. Dev Biol 37:63–89
Vertel BM, Fischman DA (1977) Mitochondrial development during myogenesis. Dev Biol 58:356–371
Yaffe D (1969) Cellular aspects of muscle differentiation in vitro. In: Moscona A, Monroy A (eds) Current topics in developmental biology. Academic Press, New York, pp 37–77
Yasmineh WG, Yunis JJ (1970) Localization of mouse satellite DNA in constitutive heterochromatin. Exp Cell Res 59:69–75
Author information
Authors and Affiliations
Additional information
Dedicated to Prof. Dr. W. Graumann on the occasion of his 65th birthday
Rights and permissions
About this article
Cite this article
Bachmann, P., Hinrichsen, K. Semiautomated evaluation of morphometric data of myoblasts and myotubes and the calculation of the relative DNA content with the television texture analysis system (TAS). Histochemistry 69, 233–242 (1980). https://doi.org/10.1007/BF00489770
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00489770