Summary
The myotome of early chick embryos was investigated histochemically by means of the acetylcholinesterase (AChE) reaction.
Light-microscopically, at the cervical level, the myotome was first recognized and AChE activity demonstrated at stage 13 (2 day-old embryo). Subsequently, the myotome elongated ventro-laterally along the inner surface of the dermomyotome and reached the ventro-lateral end of the dermomyotome at stage 17 to 18 (3 day-old embryo). AChE activity in the myotome showed subsequent increase in intensity during the course of development. The myotome consisted mainly of AChE-positive cells displaying enzymatic activity along the nuclear membrane and within the cytoplasm. In contrast, almost all cells of the dermomyotome and the interstitial cells were AChE-negative.
Electron-microscopically, the myotome cells of the 2 day-old embryo and the cells in the dorso-medial portion of the myotome of the 3 day-old embryo were morphologically undifferentiated; AChE activity was detected in the nuclear envelope and in single short profiles of the endoplasmic reticulum (ER). On the other hand, in the 3 day-old embryo the cells in the ventro-lateral portion of the myotome showed AChE activity in the nuclear envelope, numerous profiles of the ER and some Golgi complexes. These AChE-positive cells were regarded as developing myogenic cells based on their morphological characteristics.
The present findings indicate (i) that the appearance of AChE activity in the cytoplasm is the first sign of the differentiation of myogenic cells, and (ii) that in these myogenic cells the increase in AChE activity is based on the development of the ER.
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References
Allen ER, Pepe FA (1965) Ultrastructure of developing muscle cells in the chick embryo. Am J Anat 116:115–148
Atsumi S (1971) The histogenesis of motor neurons with special reference to the correlation of their endplate formation. I. The development of endplates in the intercostal muscle in the chick embryo. Acta Anat 80:161–182
Carlson BM (1981) Patten's foundations of embryology. McGraw-Hill Book Company, New York St. Louis San Francisco Auckland Bogotá Hamburg Johannesburg London Madrid Mexico Montreal New Delhi Panama Paris São Paulo Singapore Sydney Tokyo Toronto, p 279
Dalton AJ (1955) A chrome-osmium fixative for electron microscopy. Anat Rec 121:281
Dessouky DA, Hibbs RG (1965) An electron microscopic study of the development of the somatic muscle of the chick embryo. Am J Anat 116:523–566
Drews U (1975) Cholinesterase in embryonic development. Prog Histochem Cytochem 7:1–52
Engel WK (1961) Cytochemical localization of cholinesterases in cultured skeletal muscle cells. J Histochem Cytochem 9:66–72
Ezerman EB, Ishikawa H (1967) Differentiation of the sarcoplasmic reticulum and T-system in developing chick skeletal muscle in vitro. J Cell Biol 35:405–420
Fambrough D, Rash JE (1971) Development of acetylcholine sensitivity during myogenesis. Dev Biol 26:55–68
Fluck RA, Strohman RC (1973) Acetylcholinesterase activity in developing skeletal muscle in vitro. Dev Biol 33:417–428
Friedel SC, Johnson DD (1977) Neurotrophic influence on acetylcholinesterase isozymes in cultured skeletal muscle. Exp Neurol 57:257–263
Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Hay ED (1963) The fine structure of differentiating muscle in the salamander tail. Z Zellforsch mikrosk Anat 59:6–34
Hirano H (1967) A histochemical study of the cholinesterase activity in the neuromuscular junction in developing chick skeletal muscle. Arch Histol Jpn 28:89–101
Ishikawa H (1968) Formation of elaborate networks of T-system tubules in cultured skeletal muscle with special reference to the T-system formation. J Cell Biol 38:51–66
Ishikawa H (1974) Morphological analysis of muscle differentiation (in Japanese). Seitai no Kagaku 25:28–41
Ishikawa H (1976) Morphogenetic aspects of myogenesis in vitro (in Japanese). Shinkei Kenkyu no Shinpo 20:37–49
Karnovsky MJ, Roots L (1964) A “direct-coloring” thiocholine method for cholinesterases. J Histochem Cytochem 12:219–221
Khera K, Lahman QN (1965) Cholinesterases and motor endplates in developing duck skeletal muscle. J Histochem Cytochem 13:559–565
Kilarski W, Jakubowska M (1979) An electron microscope study of myofibril formation in embryonic rabbit skeletal muscle. Z Mikrosk Anat Forsch 93:1159–1181
Langman J, Nelson GR (1968) A radioautographic study of the development of the somite in the chick embryo. J Embryol Exp Morphol 19:217–226
Lewis PR, Shute CCD (1966) The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscope. J Cell Biol 1:381–390
Lewis PR, Shute CCD (1969) An electron-microscopic study of cholinesterase distribution in the rat adrenal medulla. J Microscopy 89:181–193
Linkhart TA, Hauschka SD (1979) Clonal analysis of vertebrate myogenesis IV. Acetylcholinesterase and acetylcholine receptor in myogenic and nonmyogenic clones from chick embryo leg cells. Dev Biol 69:529–548
Miki A, Atoji Y, Fujimoto E, Kikui Y, Mizoguti H (1981 a) A new method to perform autoradiography using 3H-thymidine and histochemical demonstration of acetylcholinesterase activity on the same section (in Japanese). Acta Anat Nippon 56:213–217
Miki A, Atoji Y, Mizoguti H (1981 b) The relationship between the proliferating ability and acetylcholinesterase activity of the neural tube cells in early chick embryos. Acta Histochem Cytochem 14:461–475
Millonig G (1962) Further observation on a phosphate buffer for osmium solution in fixation. Electron Microscopy Vol 2 Fifth International Congress for Electron Microscopy Academic Press, New York, p8
Mumenthaler M, Engel WK (1961) Cytological localization of cholinesterase in developing chick embryo skeletal muscle. Acta Anat 47:274–299
Okazaki K, Holtzer H (1966) Myogenesis: fusion, myosin synthesis, and the mitotic cycle. Proc Natl Acad Sci USA 56:1484–1490
Prives J, Paterson BM (1974) Differentiation of cell membranes in cultures of embryonic chick breast muscle. Proc Natl Acad Sci USA 71:3208–3211
Prives J, Silman I, Amsterdam A (1976) Appearance and disappearance of acetylcholine receptor during differentiation of chick skeletal muscle in vitro. Cell 7:543–550
Przybylski RJ, Blumberg JM (1966) Ultrastructural aspects of myogenesis in the chick. Lab Invest 15:836–863
Sawyer HR, Goldner TK, Nieberg PS, Wilson BW (1976) Ultrastructural localization of acetylcholinesterase in cultured cells. I. Embryo muscle. J Histochem Cytochem 24:969–978
Tennyson VM, Brzin M, Slotwiner P (1971) The appearance of acetylcholinesterase in the myotome of the embryonic rabbit. An electron microscope cytochemical and biochemical study. J Cell Biol 51:703–721
Tennyson VM, Brzin M, Kremzner LT (1973) Acetylcholinesterase activity in the myotube and muscle satellite cell of the fetal rabbit: an electron microscopic-cytochemical and biochemical study. J Histochem Cytochem 21:634–652
Teräväinen H (1968) Carboxylic esterases in developing myoneural junctions of rat striated muscle. Histochemie 12:307–315
Veneroni G, Murray MR (1969) Formation de novo and development of neuromuscular junctions in vitro. J Embryol Exp Morphol 21:369–382
Wake K (1964) Motor endplates in developing chick embryo skeletal muscle. Histological structure and histochemical localization of cholinesterase activity. Arch Histol Jpn 25:23–41
Wake K (1976) Formation of myoneural and myotendinous junctions in the chick embryo. Cell Tissue Res 173:383–400
Williams LW (1910) The somites of the chick. Am J Anat 11:55–100
Wilson BM, Nieberg PS, Walker CR, Linkhart TA, Fry DM (1973) Production and release of acetylcholinesterase by cultured chick embryo muscle. Dev Biol 33:285–299
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Miki, A., Mizoguti, H. Acetylcholinesterase activity in the myotome of the early chick embryo. Cell Tissue Res. 227, 23–40 (1982). https://doi.org/10.1007/BF00206329
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DOI: https://doi.org/10.1007/BF00206329