Skip to main content
Log in

Visualization of collagenase-sensitive acetylcholinesterase in isolated cardiomyocytes and in heart tissue

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

AChE :

acetylcholinesterase

BSA :

bovine serum albumin

PBS :

phosphate-buffered saline

DME :

Dulbecco's Modified Eagle Medium

References

  • Anglister L, Silman I (1978) Molecular structure of enlongated forms of electric eel acetylcholinesterase. J Mol Biol 125:293–311

    Google Scholar 

  • Anglister L, McMahan UJ (1985) Basal lamina directs acetylcholinesterase accumulation at synaptic sites in regenerating muscle. J Cell Biol 101:735–743

    Google 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 94:25–29

    Google Scholar 

  • Bon S, Vigny M, Massoulie J (1979) Asymmetric and globular forms of acetylcholinesterase in mammals and birds. Proc Natl Acad Sci USA 76:2546–2550

    Google Scholar 

  • Brimijoin S, Rakonczay Z, Mintz K (1986) Immunochemistry of mammalian cholinesterases. Fed Proc 54:2960–2964

    Google Scholar 

  • Buxton ILO, Brunton LL (1986) Compartmentation of hormone action. Adv Exp Med Biol 194:117–127

    Google Scholar 

  • Couteaux R (1955) Localization of cholinesterases at neuromuscular junction. Int Rev Cytol 4:355–375

    Google Scholar 

  • Eghbali M, Seifter S, Robinson TF, Blumenfeld OO (1987) Enzyme-antibody histochemistry: A method for detection of collagens collectively. Histochemistry 87:257–262

    Google Scholar 

  • Engval E (1980) Enzyme immunoassay ELISA and EMIT. Methods Enzymol 70:419–439

    Google Scholar 

  • Fonnum F (1969) Radiochemical microassays for the determination of choline acetyltransferase and acetylcholinesterase activities. Biochem J 115:465–472

    Google 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 133:31–38

    Google Scholar 

  • Hall Z (1973) Multiple forms of acetylcholinesterase and their distribution in endplate and non-endplate regions of rat diaphragm muscle. J Neurobiol 4:343–361

    Google Scholar 

  • Inestrosa NC, Silberstein L, Hall ZW (1982) Association of the synaptic form of acetylcholinesterase with extracellular matrix in cultured mouse muscle cells. Cell 29:71–79

    Google 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 262:4441–4444

    Google Scholar 

  • Kent MK, Cooper T (1974) The denervated heart; a model for studying antonomic control of the heart. New Engl J Med 291:1017–1021

    Google Scholar 

  • Koelle GB (1963) Cytological distributions and physiological functions of cholinesterases. In: Handbuch der experimentellen Pharmakologie, XV, Springer, Berlin Heidelberg New York, 1987

    Google 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 38:1007–1021

    Google Scholar 

  • Mark G, Chamley J, Burnstock G (1973) Interactions between autonomic nerves and smooth and cardiac muscle cells in tissue culture. Dev Biol 32:194–200

    Google 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 74:3073–3077

    Google Scholar 

  • Massoulie J, Bon S (1982) The molecular forms of cholinesterase and acetylcholinesterase in vertebrates. Ann Rev Neurosci 5:57–106

    Google Scholar 

  • Purves RD, Hill CE, Chamley J, Mark GE, Fry DM, Burnstock G (1974) Functional autonomic neuromuscular junctions in tissue culture. Pflügers Arch 350:1–7

    Google 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 16:3550–3558

    Google Scholar 

  • Sandusky GE, White SL, Wightman KA (1986) Canine artioventricular node: Scanning electron microscopy and enzyme histochemistry. Am J Vet Res 47:304–308

    Google Scholar 

  • Silman I, Futerman H (1987) Post-translational modification as a means of anchoring acetylcholinesterase to the cell surface. Biopolymers 26:S241-S253

    Google Scholar 

  • Silman I, Lyles JM, Barnard EA (1978) Intrinsic forms of acetylcholinesterase in skeletal muscle. FEBS Lett 94:166–170

    Google Scholar 

  • Silver A (1974) In: Neuberger A, Tatum EL (eds) The biology of cholinesterase in frontiers of biology. North-Holland, Amsterdam

    Google Scholar 

  • Slavikova J, Ylk J, Hlavickova Y (1982) Acetylcholinesterase and butyrylcholinesterase activity in the atria of the heart of adult albino rats. Physiol Bohemoslov 31:407–417

    Google Scholar 

  • Torigoe K, Nakamura T (1987) Fine structure of myomyous junctions in the mouse skeletal muscles. Tissue Cell 19:243–250

    Google 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 27:1347–1353

    Google Scholar 

  • Vigny M, Martin GR, Grotendorst GR (1983) Interactions of asymmetric forms of acetylcholinesterase with basement membrane components. J Biol Chem 258:8794–8798

    Google 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 216:231–251

    Google 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 59:143–150

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eghbali, M., Silman, I., Robinson, T.F. et al. Visualization of collagenase-sensitive acetylcholinesterase in isolated cardiomyocytes and in heart tissue. Cell Tissue Res. 253, 281–286 (1988). https://doi.org/10.1007/BF00222282

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00222282

Key words

Navigation