Abstract
Confocal laser-scanning microscopy of phalloidine-stained actin fibers is a relatively new tool for studying the development of myocardial fiber organization. It seems to show orientation of myocytes in rather early embryonic stages. To further evaluate the differentiation of the myocardium, this technique was compared with transmission electron microscopy in rat embryos aged between 11 and 18 days. Although the confocal images of actin filament patterns pointed to early myocyte orientation, the electron micrographs revealed that even at 17 days the ventricular myocardium was far from mature. Myofibrils never completely filled the myocytes, and lack of organization was the rule rather than the exception. The organized structure as revealed by confocal microscopy was based on cell-to-cell continuity, whereas electron microscopy showed crossing and disarray within individual myocytes. Exceptions were in the ventricular trabeculations, which showed precocious myofiber differentiation. The trabeculations probably support ventricular systole in those stages in which the free walls do not yet provide efficient contractions. The other exception was the wall of the outflow tract, which showed well-oriented myofibrils from early stages onwards. Apparently, the outflow tract has a different function in these stages. The differences found between confocal microscopy and electron microscopy suggest that some caution is indicated in the interpretation of fluorescent images of relatively low magnification.
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References
Challice CE, Virágh S (1973) The architectural development of the early mammalian heart. Tissue Cell 6:447–462
Clark EB, Hu N, Dummett JL, Vandekieft GK, Olson C, Tomanek R (1986) Ventricular function and morphology in chick embryo from stages 18 and 29. Am J Physiol 250:H407-H413
DeJong F, Opthof T, Wilde AAM, Janse MJ, Charles R, Lamers WH, Moorman AFM (1992) Persisting zones of slow impulse conduction in developing chicken hearts. Circ Res 71:240–250
Fawcett DW, McNutt NS (1969) The ultrastructure of the cat myocardium. I. Ventricular papillary muscle. J Cell Biol 42:1–45
Fox CC, Hutchins GM (1972) The architecture of human ventricular myocardium. Johns Hopkins Med J 130:289–299
Fuseler JW, Shay JW, Feit H (1981) The role of intermediate (10 nm) filaments in the development and integration of myofibrillar contractile apparatus in the embryonic mammalian heart. In: Dowben RM, Shay JW (eds) Cell and muscle motility, vol I. Plenum Press, New York London, pp 205–259
Goerttler K (1955) Über Blutstromwirkung als Gestaltungsfaktor für die Entwicklung des Herzens. Beitr Pathol Anat Allg Pathol 115:33–56
Ingber DE, Jamieson JD (1985) Cells as tensegrity structures: architectural regulation of histodifferentiation by physical forces transduced over basement membrane. In: Anderson LC, Gahmberg CG, Eckblom P (eds) Gene expression during normal and malignant differentiation. Academic Press, London, pp 13–23
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 the caudal part of the heart tube play a role in dextro-looping in the embryonic chick heart. Anat Embryol 183:29–39
Jaffee OC (1965) Hemodynamic factors in the development of the chicken embryo heart. Anat Rec 151:69–76
Jouk P-S, Usson Y, Michalowicz G, Parazza F (1995) Mapping of the orientation of myocardial cells by means of polarized light and confocal scanning laser microscopy. Microsc Res Tech 30:480–490
Karnovsky MJ (1965) A formaldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27:137A
Knaapen MWM, Vrolijk BCM, Wenink ACG (1995a) The growth of the individual segments in the embryonic rat heart. Ann NY Acad Sci 752:105–107
Knaapen MWM, Vrolijk BCM, Wenink ACG (1995b) Growth of the myocardial volumes of the individual cardiac segments in the rat embryo. Anat Rec 243:93–100
Knaapen MWM, Vrolijk BCM, Wenink ACG (1996) Cellular distribution of myofibrils, mitochondria and mitochondrial cristae. In: Knaapen MWM. Differentiation of the myocardium in the embryonic rat heart. Quantitative information to support qualitative changes. Thesis, Leiden.
Komoiyama M, Kouchi K, Maruyama K, Shimada Y (1993) Dynamics of actin and assembly of connectin (titin) during myofibrillogenesis in embryonic chick cardiac muscle cells in vitro. Dev Dyn 196:291–299
Kouchi K, Takahashi H, Shimada Y (1993) Incorporation of biotin-labelled actin into nascent myofibrils of cardiac myocytes: an immunoelectron microscopic study. J Muscle Res Cell Motil 14:292–301
Lacktis JW, Manasek FJ (1978) An analysis of deformation during a normal morphogenetic event. In: Rosenquist GC, Bergsma D (eds) Morphogenesis and malformation of the cardiovascular system. Birth defects, original article series XIV-7. Liss, New York, pp 205–227
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 (1970) Histogenesis of the embryonic myocardium. Am J Cardiol 25:149–168
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, Mount Kisco, New York, pp 359–376
Manasek J (1976) Heart development: interactions involved in cardiac morphogenesis. In: Poste G, Nicolson GL (eds) The cell surface in animal embryogenesis and development. North-Holland, Amsterdam New York Oxford, pp 545–598
Maron BJ, Sato N, Roberts WC, Edwards JE, Chandra RS (1979) Quantitative analysis of cardiac muscle cell disorganization in the ventricular septum. Comparison of fetuses and infants with and without congenital heart disease and patients with hypertrophic cardiomyopathy. Circulation 60:685–696
Nakamura A, Kulikowski RR, Lacktis JW, Manasek FJ (1980) Heart looping: a regulated response to deforming forces. In: Van Praagh R, Takao A (eds) Etiology and morphogenesis of congenital heart disease. Futura, Mount Kisco, New York, pp 81–98
Ostadal B, Schiebler TH, Rychter Z (1988) Relations between development of the capillary wall and myo-architecture of the rat heart. In: Skalak R, Fox CF (eds) UCLA Symposium on molecular and cellular biology. Liss, New York, pp 375–388
Sanchez-Quintana D, Climent V, Garcia-Martinez V, Rojo M, Hurle JM (1994) Spatial arrangement of the heart muscle fascicles and intramyocardial connective tissue in the Spanish fighting bull (Bos taurus). J Anat 184:273–283
Shiraishi I, Takamatsu T, Minimikawa T, Fujita S (1992) 3D-observation of actin filaments during cardiac myofibrinogenesis in chick embryo using a confocal laser scanning microscope. Anat Embryol 185:401–408
Streeter DD (1979) Gross morphology and geometry of the heart. In: Berne RM (ed) The cardiovascular system, vol I. The heart. Am Physiol Soc, Bethesda, pp 61–112
Taber LA, Sun H, Clark EB, Keller BB (1994) Epicardial strains in embryonic chick ventricle at stages 16 through 24. Circ Res 75:896–903
Tandler J (1913) Anatomie des Herzens. Fischer, Jena
Terracio L, Peters W, Durig B, Miller B, Borg K, Borg TK (1988) Cellular hypertrophy can be induced by cyclical mechanical stretch in vitro. In: Skalak R, Fox CF (eds) UCLA Symposium on molecular and cellular biology. Liss, New York, pp 1–6
Tokuyasu KT (1980) Co-development of embryonic myocardium and myocardial circulation. In: Clark EB, Takao A (eds) Developmental cardiology: Morphogenesis and function. Futura, Mount Kisco, New York, pp 205–218
Usson Y, Parazza F, Jouk P-S, Michalowicz G (1994) Method for the study of the three-dimensional orientation of the nuclei of myocardial cells in fetal human heart by means of confocal scanning laser microscopy. J Microsc 174:101–110
VanGroningen JP, Wenink ACG, Testers LHM (1991) Myocardial capillaries: increase in number by splitting of existing vessels. Anat Embryol 184:65–70
Virágh S, Challice CE (1973) Origin and differentiation of cardiac muscle cells in the mouse. J Ultrastruct Res 42:1–24
Virágh S, Challice CE (1981) The origin of the epicardium and the embryonic myocardial circulation in the mouse. Anat Rec 201:157–168
Vrolijk BCM, Knaapen MWM, Wenink ACG (1995) Technical possibilities to obtain random ultrathin sections. Eur J Morphol 33:299–301
Wenink ACG, Spliet WGM, Mansoer JR (1988) Entwicklung der räumlichen Anordnung des Myokardiums. Wien Klin Wochenschr 110:805–811
Zak R (1981) Contractile function as a determinant of muscle growth. In: Dowben RM, Shay JW (eds) Cell and muscle motility, vol I. Plenum Press, New York London, pp 1–33
Zak R, Kizu A, Bugaisky L (1979) Cardiac hypertrophy: its characteristics as a growth process. Am J Cardiol 44:941–946
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Wenink, A.C.G., Knaapen, M.W.M., Vrolijk, B.C.M. et al. Development of myocardial fiber organization in the rat heart. Anat Embryol 193, 559–567 (1996). https://doi.org/10.1007/BF00187927
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DOI: https://doi.org/10.1007/BF00187927