Summary
Using a confocal laser scanning microscope (CLSM), we observed subcellular three-dimensional (3-D) arrangements of actin filaments stained with fluorescein-labeled phalloidin during myofibrinogenesis of chick embryonic heart (7- to 13-somite stages). Serial optical tomograms were obtained from whole-mounted heart tubes and reconstructed into stereoscopic images. Development of myofibrils in myocardial differentiation considerably differed in inner and outer myocardial cell layers. In the outer layer, initial myofibrils appeared along cell membranes at the 8-somite stage. They increased rapidly and constituted network structures with spatial extension over cell-cell junctions. In the inner layer, myofibrils appeared at the bottom, facing the cardiac jelly, at the 10-somite stage, and, when the straight heart tube began to bend, they were already aligned circumferentially in the direction of the heart tube. Double staining of fluorescein-phalloidin and DiI [1,1′-dioctadecyl-3, 3,3′,3′-tetramethylindo-carbocyanine perchlorate; DiI-C18-(3)] of the looped heart revealed that while myocytes in the outer layer were round, those of the inner layer were spindle-shaped, and their long axes coincided with the circumferential direction. These results suggest that the circumferentially arranged myofibrils at the bottom of the inner layer may play an important role in the looping of the heart tube.
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References
Borg TK, Xuehui M, Hilenski L, Vinson N, Terracio L (1990) The role of the extracellular matrix on myofibrinogenesis in vitro. In: Clark E, Takao A (eds) Developmental cardiology: morphogenesis and function. Futura, New York, pp 175–190
Dlugosz AA, Antin PB, Nachmias VT, Holtzer H (1984) The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes. J Cell Biol 99:2268–2278
Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Hiruma T, Hirakow R (1985) An ultrastructural topographical study on myofibrinogenesis in the heart of the chick embryo during first onset period. Anat Embryol 172:325–329
Honig MG, Hume RI (1986) Fluorescent carbocyanine dyes allow living neurons of identified origin to be studied in long-term cultures. J Cell Biol 103:171–187
Itasaki N, Nakamura H, Yasuda M (1989) Changes in the arrangement of actin bundles during heart looping in the chick embryo. Anat Embryol 180:413–420
Itasaki N, Nakamura H, Sumida H, Yasuda M (1991) Actin bundles on the right side in caudal part of the heart tube play a role in dextro-looping in the embryonic chick heart. Anat Embryol 183:29–39
Linder E (1960) Myofibrils in the early development of chick embryo hearts as observed with the electron microscope. Anat Rec 36:234–235
Little CD, Piquet DM, Davis LA, Walters L, Drake CJ (1989) Distribution of laminin, collagen type IV, collagen type I, and fibronectin in chicken cardiac jelly/basement membrane. Anat Rec 224:417–425
Manasek FJ (1968) Embryonic development of the heart. I. A light and electron microscopic study of myocardial development in the early chick embryo. J Morphol 125:329–366
Manasek FJ (1981) Determinations of heart shape in early embryos. Fed Proc 40:2011–2016
Manasek FJ, Isobe Y, Shimada Y, Hopkins W (1984) The embryonic myocardial cytoskeleton, interstitial pressure, and the control of morphogenesis. In: Nora JJ, Takao A (eds) Congenital heart disease: causes and processes. Futura, New York, pp 359–376
Minamikawa T, Takamatsu T, Fujita S (1991) Application of laser microtomography of in situ three dimensional subcellular morphology. Acta Histochem Cytochem 24:55–60
Nagafuchi A, Takeichi M (1988) Cell biding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J 7:3679–3684
Sanger JM, Mittal B, Pochapin MB, Sanger JW (1986) Myofibrinogenesis in living cells microinjected with fluorescently labeled alpha-actinin. J Cell Biol 102:2053–2066
Stalsberg H (1969a) The origin of heart asymmetry: right and left contributions to the early chick embryo heart. Dev Biol 19:109–127
Stalsberg H (1969b) Regional mitotic activity in the precardiac mesoderm and differentiating heart tube in the chick embryo. Dev Biol 20:18–45
Takamatsu T, Fujita S (1987) Confocal laser scanning microscopy and its three dimensional application. J Microsc 149:167–174
Tokuyasu KT (1987) Immunohistochemical studies of cardiac myofibrinogenesis in early chick embryos. II. Generation of alphaactinin dots within titin spots at the time of the first myofibril formation. J Cell Biol 105:2795–2801
Tokuyasu KT (1989) Immunohistochemical Studies of cardiac myofibrinogenesis in early chick embryos. III. Generation of fasciae adherents and costameres. J Cell Biol 108:43–53
Tokuyasu KT (1990) Co-development of embryonic myocardium and myocardial circulation. In: Clark E, Takao A (eds) Developmental cardiology: morphogenesis and function. Futura, New York, pp 201–218
Tokuyasu KT, Maher PA (1987) Immunocytochemical studies of cardiac myofibrinogenesis in early chick embryos. I. Presence of immunofluorescent titin spots and in the pre-myofibril stages. J Cell Biol 105:2781–2793
Volk T, Geiger B (1986) A-CAM: A 135-kD receptor of intercellular adherens junctions. II. Antibody-mediated modulation of junction formation. J Cell Biol 103:1451–1464
Wilens S (1955) The migration of heart mesoderm and associated areas in Amblystoma punctatum. J Exp Zool 129:579–605
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Shiraishi, I., Takamatsu, T., Minamikawa, T. et al. 3-D observation of actin filaments during cardiac myofibrinogenesis in chick embryo using a confocal laser scanning microscope. Anat Embryol 185, 401–408 (1992). https://doi.org/10.1007/BF00188551
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DOI: https://doi.org/10.1007/BF00188551