Visualization of collagenase-sensitive acetylcholinesterase in isolated cardiomyocytes and in heart tissue
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Summary
Previous studies have indicated that the asymmetric form of acetylcholinesterase (collagen-tailed) is localized in the basal lamina of the neuromuscular junction of skeletal muscle. The present study shows localization of the asymmetric acetylcholinesterase in the heart of the rat. Antiserum to 14+18 S acetylcholinesterase of the electric eel was raised in rabbits. The purified antibody did not react with collagen type I or laminin. Collagenase reduced the immunoreactivity of the enzyme with the purified antibody. Isolated cardiomyocytes and frozen sections of the heart were stained for acetylcholinesterase with the antibody. Diffuse immunofluorescence appeared over the surface of the cardiomyocytes. In the frozen sections, the immunofluorescence was most intense at the cell boundaries. These data suggest that collagenase-sensitive acetylcholinesterase in the heart is present in the myocytes and occurs in the vicinity of the basal lamina.
Key words
Cardiomyocytes Collagen-tailed acetylcholinesterase Basal lamina Myocardium Rat (Fischer 344)Abbreviations
- AChE
acetylcholinesterase
- BSA
bovine serum albumin
- PBS
phosphate-buffered saline
- DME
Dulbecco's Modified Eagle Medium
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References
- Anglister L, Silman I (1978) Molecular structure of enlongated forms of electric eel acetylcholinesterase. J Mol Biol 125:293–311Google Scholar
- Anglister L, McMahan UJ (1985) Basal lamina directs acetylcholinesterase accumulation at synaptic sites in regenerating muscle. J Cell Biol 101:735–743Google Scholar
- Anglister L, Tarrab-Hazdai R, Fuchs S, Silman I (1978) Immunological cross-reactivity between electric eel acetylcholinesterase and rat tail tendon collagen. Eur J Biochem 94:25–29Google Scholar
- Bon S, Vigny M, Massoulie J (1979) Asymmetric and globular forms of acetylcholinesterase in mammals and birds. Proc Natl Acad Sci USA 76:2546–2550Google Scholar
- Brimijoin S, Rakonczay Z, Mintz K (1986) Immunochemistry of mammalian cholinesterases. Fed Proc 54:2960–2964Google Scholar
- Buxton ILO, Brunton LL (1986) Compartmentation of hormone action. Adv Exp Med Biol 194:117–127Google Scholar
- Couteaux R (1955) Localization of cholinesterases at neuromuscular junction. Int Rev Cytol 4:355–375Google Scholar
- Eghbali M, Seifter S, Robinson TF, Blumenfeld OO (1987) Enzyme-antibody histochemistry: A method for detection of collagens collectively. Histochemistry 87:257–262Google Scholar
- Engval E (1980) Enzyme immunoassay ELISA and EMIT. Methods Enzymol 70:419–439Google Scholar
- Fonnum F (1969) Radiochemical microassays for the determination of choline acetyltransferase and acetylcholinesterase activities. Biochem J 115:465–472Google Scholar
- Grassi J, Massoulie J, Timpl R (1983) Relationship of collagentailed acetylcholinesterase with basal lamina components: Absence of binding with laminin, fibronectin, and collagen typesIV and V and lack of reactivity with different anti-collagen sera. Eur J Biochem 133:31–38Google Scholar
- Hall Z (1973) Multiple forms of acetylcholinesterase and their distribution in endplate and non-endplate regions of rat diaphragm muscle. J Neurobiol 4:343–361Google Scholar
- Inestrosa NC, Silberstein L, Hall ZW (1982) Association of the synaptic form of acetylcholinesterase with extracellular matrix in cultured mouse muscle cells. Cell 29:71–79Google Scholar
- Inestrosa NC, Roberts WL, Marshall TL, Rosenberry TL (1987) Acetylcholinesterase from bovine caudate nucleus is attached to membranes by a novel subunit distinct from those of acetylcholinesterases in other tissues. J Biol Chem 262:4441–4444Google Scholar
- Kent MK, Cooper T (1974) The denervated heart; a model for studying antonomic control of the heart. New Engl J Med 291:1017–1021Google Scholar
- Koelle GB (1963) Cytological distributions and physiological functions of cholinesterases. In: Handbuch der experimentellen Pharmakologie, XV, Springer, Berlin Heidelberg New York, 1987Google Scholar
- Lyles JM, Silman I, DiGiamberardino L, Couraud JY, Barnard EA (1981) Comparison of the molecular forms of the cholinesterases in tissues of normal and dystrophic chickens. J Neurochem 38:1007–1021Google Scholar
- Mark G, Chamley J, Burnstock G (1973) Interactions between autonomic nerves and smooth and cardiac muscle cells in tissue culture. Dev Biol 32:194–200Google Scholar
- Marshall LM, Sanes JR, Memahan UJ (1977) Reinnervation of original synaptic sites on muscle fiber basement membrane after disruption of the muscle cells. Proc Natl Acad Sci USA 74:3073–3077Google Scholar
- Massoulie J, Bon S (1982) The molecular forms of cholinesterase and acetylcholinesterase in vertebrates. Ann Rev Neurosci 5:57–106Google Scholar
- Purves RD, Hill CE, Chamley J, Mark GE, Fry DM, Burnstock G (1974) Functional autonomic neuromuscular junctions in tissue culture. Pflügers Arch 350:1–7Google Scholar
- Rosenberry TL, Richardson JM (1977) Structure of 18 S and 14 S acetylcholinesterase. Identification of collagen-like subunits that are linked by disulfide bonds to catalytic subunits. Biochemistry 16:3550–3558Google Scholar
- Sandusky GE, White SL, Wightman KA (1986) Canine artioventricular node: Scanning electron microscopy and enzyme histochemistry. Am J Vet Res 47:304–308Google Scholar
- Silman I, Futerman H (1987) Post-translational modification as a means of anchoring acetylcholinesterase to the cell surface. Biopolymers 26:S241-S253Google Scholar
- Silman I, Lyles JM, Barnard EA (1978) Intrinsic forms of acetylcholinesterase in skeletal muscle. FEBS Lett 94:166–170Google Scholar
- Silver A (1974) In: Neuberger A, Tatum EL (eds) The biology of cholinesterase in frontiers of biology. North-Holland, AmsterdamGoogle Scholar
- Slavikova J, Ylk J, Hlavickova Y (1982) Acetylcholinesterase and butyrylcholinesterase activity in the atria of the heart of adult albino rats. Physiol Bohemoslov 31:407–417Google Scholar
- Torigoe K, Nakamura T (1987) Fine structure of myomyous junctions in the mouse skeletal muscles. Tissue Cell 19:243–250Google Scholar
- Vigny M, Koenig J, Rieger F (1976) The motor endplate specific form of acetylcholinesterase: Appearance during embryogenesis and reinnervation of rat muscle. J Neurochem 27:1347–1353Google Scholar
- Vigny M, Martin GR, Grotendorst GR (1983) Interactions of asymmetric forms of acetylcholinesterase with basement membrane components. J Biol Chem 258:8794–8798Google Scholar
- Wittenberg BA, Robinson TF (1981) Oxygen requirements, morphology, cell coat, and membrane permeability of calcium-tolerant myocytes from hearts of adult rats. Cell Tissue Res 216:231–251Google Scholar
- Wittenberg BA, White RL, Ginzberg RD, Spray DC (1986) Effect of calcium on the dissociation of the mature rat heart into individual paired myocytes: Electrical properties of cell pairs. Circ Res 59:143–150Google Scholar