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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1990 Oct;86(4):1293–1300. doi: 10.1172/JCI114837

Myocardial localization and isoforms of neural cell adhesion molecule (N-CAM) in the developing and transplanted human heart.

L Gordon 1, J Wharton 1, S E Moore 1, F S Walsh 1, J G Moscoso 1, R Penketh 1, J Wallwork 1, K M Taylor 1, M H Yacoub 1, J M Polak 1
PMCID: PMC296861  PMID: 2212013

Abstract

Neural cell adhesion molecule (N-CAM) has been implicated in cellular interactions involved in cardiac morphogenesis and innervation. Immunohistochemical techniques and Western blot analysis were used to determine the localization and isoforms of N-CAM in the developing and extrinsically denervated human heart. Myocardial and conducting cells in the fetal heart (7-24 wk gestation) exhibited sarcolemmal immunoreactivity, the major desialo N-CAM isoforms being 150, 145, 120, 115, and 110 kD. N-CAM expression appeared to be downregulated in the myocardium during adult life, with relatively little sarcolemmal immunoreactivity being detected in normal donor tissues. In contrast to the temporal changes observed in the myocardium, both the developing and mature cardiac innervation displayed N-CAM immunofluorescence staining, localized to neuronal cell bodies, nerve fascicles and fibres. Extrinsically denervated cardiac allografts, obtained 2 d to 91 mo after transplantation, showed extensive sarcolemmal and intercalated disc immunostaining and expression of 125-, 120-, and 115-kD isoforms. Tissues from explanted recipient hearts and atrial appendage samples obtained during coronary bypass graft operations were also examined and displayed varying amounts of N-CAM immunoreactivity. We conclude that the expression of N-CAM immunoreactivity and isoforms in the human heart is developmentally regulated and may be modulated by factors such as cardiac innervation and myocardial hypertrophy.

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Selected References

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  1. Barbas J. A., Chaix J. C., Steinmetz M., Goridis C. Differential splicing and alternative polyadenylation generates distinct NCAM transcripts and proteins in the mouse. EMBO J. 1988 Mar;7(3):625–632. doi: 10.1002/j.1460-2075.1988.tb02856.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barton C. H., Dickson G., Gower H. J., Rowett L. H., Putt W., Elsom V., Moore S. E., Goridis C., Walsh F. S. Complete sequence and in vitro expression of a tissue-specific phosphatidylinositol-linked N-CAM isoform from skeletal muscle. Development. 1988 Sep;104(1):165–173. doi: 10.1242/dev.104.1.165. [DOI] [PubMed] [Google Scholar]
  3. Booth C. M., Brown M. C. Localization of neural cell adhesion molecule in denervated muscle to both the plasma membrane and extracellular compartments by immuno-electron microscopy. Neuroscience. 1988 Nov;27(2):699–709. doi: 10.1016/0306-4522(88)90299-0. [DOI] [PubMed] [Google Scholar]
  4. Covault J., Cunningham J. M., Sanes J. R. Neurite outgrowth on cryostat sections of innervated and denervated skeletal muscle. J Cell Biol. 1987 Dec;105(6 Pt 1):2479–2488. doi: 10.1083/jcb.105.6.2479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Covault J., Merlie J. P., Goridis C., Sanes J. R. Molecular forms of N-CAM and its RNA in developing and denervated skeletal muscle. J Cell Biol. 1986 Mar;102(3):731–739. doi: 10.1083/jcb.102.3.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Covault J., Sanes J. R. Neural cell adhesion molecule (N-CAM) accumulates in denervated and paralyzed skeletal muscles. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4544–4548. doi: 10.1073/pnas.82.13.4544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daniloff J. K., Levi G., Grumet M., Rieger F., Edelman G. M. Altered expression of neuronal cell adhesion molecules induced by nerve injury and repair. J Cell Biol. 1986 Sep;103(3):929–945. doi: 10.1083/jcb.103.3.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dickson G., Gower H. J., Barton C. H., Prentice H. M., Elsom V. L., Moore S. E., Cox R. D., Quinn C., Putt W., Walsh F. S. Human muscle neural cell adhesion molecule (N-CAM): identification of a muscle-specific sequence in the extracellular domain. Cell. 1987 Sep 25;50(7):1119–1130. doi: 10.1016/0092-8674(87)90178-4. [DOI] [PubMed] [Google Scholar]
  9. Doherty P., Barton C. H., Dickson G., Seaton P., Rowett L. H., Moore S. E., Gower H. J., Walsh F. S. Neuronal process outgrowth of human sensory neurons on monolayers of cells transfected with cDNAs for five human N-CAM isoforms. J Cell Biol. 1989 Aug;109(2):789–798. doi: 10.1083/jcb.109.2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edelman G. M. Cell adhesion molecules in neural histogenesis. Annu Rev Physiol. 1986;48:417–430. doi: 10.1146/annurev.ph.48.030186.002221. [DOI] [PubMed] [Google Scholar]
  11. Filip D. A., Radu A., Simionescu M. Interstitial cells of the heart valves possess characteristics similar to smooth muscle cells. Circ Res. 1986 Sep;59(3):310–320. doi: 10.1161/01.res.59.3.310. [DOI] [PubMed] [Google Scholar]
  12. Gower H. J., Barton C. H., Elsom V. L., Thompson J., Moore S. E., Dickson G., Walsh F. S. Alternative splicing generates a secreted form of N-CAM in muscle and brain. Cell. 1988 Dec 23;55(6):955–964. doi: 10.1016/0092-8674(88)90241-3. [DOI] [PubMed] [Google Scholar]
  13. Grumet M., Rutishauser U., Edelman G. M. Neural cell adhesion molecule is on embryonic muscle cells and mediates adhesion to nerve cells in vitro. Nature. 1982 Feb 25;295(5851):693–695. doi: 10.1038/295693a0. [DOI] [PubMed] [Google Scholar]
  14. Hoffman S., Sorkin B. C., White P. C., Brackenbury R., Mailhammer R., Rutishauser U., Cunningham B. A., Edelman G. M. Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J Biol Chem. 1982 Jul 10;257(13):7720–7729. [PubMed] [Google Scholar]
  15. Hurko O., Walsh F. S. Human fetal muscle-specific antigen is restricted to regenerating myofibers in diseased adult muscle. Neurology. 1983 Jun;33(6):737–743. doi: 10.1212/wnl.33.6.737. [DOI] [PubMed] [Google Scholar]
  16. Ibsen S., Berezin V., Nørgaard-Pedersen B., Bock E. Enzyme-linked immunosorbent assay of the D2-glycoprotein. J Neurochem. 1983 Aug;41(2):356–362. doi: 10.1111/j.1471-4159.1983.tb04750.x. [DOI] [PubMed] [Google Scholar]
  17. Imakita M., Tazelaar H. D., Rowan R. A., Masek M. A., Billingham M. E. Myocyte hypertrophy in the transplanted heart. A morphometric analysis. Transplantation. 1987 Jun;43(6):839–842. [PubMed] [Google Scholar]
  18. Kondo Y., Matheny J. L., Hardy J. D. Autonomic reinnervation of cardiac transplants: further observations in dogs and rhesus monkeys. Ann Surg. 1972 Jul;176(1):42–48. doi: 10.1097/00000658-197207000-00009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kontos H. A., Thames M. D., Lower R. R. Responses to electrical and reflex autonomic stimulation in dogs with cardiac transplantation before and after reinnervation. J Thorac Cardiovasc Surg. 1970 Mar;59(3):382–392. [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. McNutt N. S. Ultrastructure of intercellular junctions in adult and developing cardiac muscle. Am J Cardiol. 1970 Feb;25(2):169–183. doi: 10.1016/0002-9149(70)90577-1. [DOI] [PubMed] [Google Scholar]
  22. Mirsky R., Jessen K. R., Schachner M., Goridis C. Distribution of the adhesion molecules N-CAM and L1 on peripheral neurons and glia in adult rats. J Neurocytol. 1986 Dec;15(6):799–815. doi: 10.1007/BF01625196. [DOI] [PubMed] [Google Scholar]
  23. Moore S. E., Thompson J., Kirkness V., Dickson J. G., Walsh F. S. Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines. J Cell Biol. 1987 Sep;105(3):1377–1386. doi: 10.1083/jcb.105.3.1377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moore S. E., Walsh F. S. Specific regulation of N-CAM/D2-CAM cell adhesion molecule during skeletal muscle development. EMBO J. 1985 Mar;4(3):623–630. doi: 10.1002/j.1460-2075.1985.tb03675.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murray B. A., Owens G. C., Prediger E. A., Crossin K. L., Cunningham B. A., Edelman G. M. Cell surface modulation of the neural cell adhesion molecule resulting from alternative mRNA splicing in a tissue-specific developmental sequence. J Cell Biol. 1986 Oct;103(4):1431–1439. doi: 10.1083/jcb.103.4.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pinto J. E., Nazarali A. J., Torda T., Saavedra J. M. Autoradiographic characterization of beta-adrenoceptors in rat heart valve leaflets. Am J Physiol. 1989 Mar;256(3 Pt 2):H821–H827. doi: 10.1152/ajpheart.1989.256.3.H821. [DOI] [PubMed] [Google Scholar]
  27. Prediger E. A., Hoffman S., Edelman G. M., Cunningham B. A. Four exons encode a 93-base-pair insert in three neural cell adhesion molecule mRNAs specific for chicken heart and skeletal muscle. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9616–9620. doi: 10.1073/pnas.85.24.9616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Prentice H. M., Moore S. E., Dickson J. G., Doherty P., Walsh F. S. Nerve growth factor-induced changes in neural cell adhesion molecule (N-CAM) in PC12 cells. EMBO J. 1987 Jul;6(7):1859–1863. doi: 10.1002/j.1460-2075.1987.tb02444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rieger F., Nicolet M., Pinçon-Raymond M., Murawsky M., Levi G., Edelman G. M. Distribution and role in regeneration of N-CAM in the basal laminae of muscle and Schwann cells. J Cell Biol. 1988 Aug;107(2):707–719. doi: 10.1083/jcb.107.2.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rutishauser U. Developmental biology of a neural cell adhesion molecule. Nature. 1984 Aug 16;310(5978):549–554. doi: 10.1038/310549a0. [DOI] [PubMed] [Google Scholar]
  31. Rutishauser U., Grumet M., Edelman G. M. Neural cell adhesion molecule mediates initial interactions between spinal cord neurons and muscle cells in culture. J Cell Biol. 1983 Jul;97(1):145–152. doi: 10.1083/jcb.97.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rutishauser U., Hoffman S., Edelman G. M. Binding properties of a cell adhesion molecule from neural tissue. Proc Natl Acad Sci U S A. 1982 Jan;79(2):685–689. doi: 10.1073/pnas.79.2.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thiery J. P., Duband J. L., Rutishauser U., Edelman G. M. Cell adhesion molecules in early chicken embryogenesis. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6737–6741. doi: 10.1073/pnas.79.21.6737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thompson J., Dickson G., Moore S. E., Gower H. J., Putt W., Kenimer J. G., Barton C. H., Walsh F. S. Alternative splicing of the neural cell adhesion molecule gene generates variant extracellular domain structure in skeletal muscle and brain. Genes Dev. 1989 Mar;3(3):348–357. doi: 10.1101/gad.3.3.348. [DOI] [PubMed] [Google Scholar]
  35. Thompson J., Moore S. E., Walsh F. S. Thyroid hormones regulate expression of the neural cell adhesion molecule in adult skeletal muscle. FEBS Lett. 1987 Jul 13;219(1):135–138. doi: 10.1016/0014-5793(87)81205-x. [DOI] [PubMed] [Google Scholar]
  36. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Walsh F. S., Dickson G., Moore S. E., Barton C. H. Unmasking N-CAM. Nature. 1989 Jun 15;339(6225):516–516. doi: 10.1038/339516a0. [DOI] [PubMed] [Google Scholar]
  38. Walsh F. S., Ritter M. A. Surface antigen differentiation during human myogenesis in culture. Nature. 1981 Jan 1;289(5793):60–64. doi: 10.1038/289060a0. [DOI] [PubMed] [Google Scholar]
  39. Wharton J., Gordon L., Walsh F. S., Flanigan T. P., Moore S. E., Polak J. M. Neural cell adhesion molecule (N-CAM) expression during cardiac development in the rat. Brain Res. 1989 Mar 27;483(1):170–176. doi: 10.1016/0006-8993(89)90050-4. [DOI] [PubMed] [Google Scholar]
  40. Wharton J., Gulbenkian S., Merighi A., Kuhn D. M., Jahn R., Taylor K. M., Polak J. M. Immunohistochemical and ultrastructural localisation of peptide-containing nerves and myocardial cells in the human atrial appendage. Cell Tissue Res. 1988 Oct;254(1):155–166. doi: 10.1007/BF00220029. [DOI] [PubMed] [Google Scholar]
  41. Wharton J., Polak J. M., Gordon L., Banner N. R., Springall D. R., Rose M., Khagani A., Wallwork J., Yacoub M. H. Immunohistochemical demonstration of human cardiac innervation before and after transplantation. Circ Res. 1990 Apr;66(4):900–912. doi: 10.1161/01.res.66.4.900. [DOI] [PubMed] [Google Scholar]

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