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. 1982 Jan;79(2):665–669. doi: 10.1073/pnas.79.2.665

Immunocytochemistry with internally labeled monoclonal antibodies.

A C Cuello, J V Priestley, C Milstein
PMCID: PMC345807  PMID: 6177000

Abstract

One of the advantages of the production of monoclonal antibodies by tissue culture methods is that they can be internally labeled by using appropriate radioactive amino acid in the culture fluid. Thus, radioactive immunological probes of high specific activity can be prepared. Here we report applications of these internally labeled monoclonal antibodies for the direct localization of immunoreactive sites in the central nervous system of the rat at both light and electron microscopic levels (radioimmunocytochemistry). We explored the combined use of radioimmunocytochemistry and immunoenzymatic methods for the simultaneous detection of two antigenic sites: substance P and serotonin or substance P and enkephalin. In neurobiology this procedure could help to clarify certain aspects of transmitter-specific synaptic interactions and the coexistence of neuroactive substances in single neuronal cell bodies or nerve terminals. We also describe the application of radioimmunocytochemistry with internally labeled monoclonal antibodies to quantify the immunoreactions in discrete microscopic areas.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barber R. P., Vaughn J. E., Slemmon J. R., Salvaterra P. M., Roberts E., Leeman S. E. The origin, distribution and synaptic relationships of substance P axons in rat spinal cord. J Comp Neurol. 1979 Mar 15;184(2):331–351. doi: 10.1002/cne.901840208. [DOI] [PubMed] [Google Scholar]
  2. Chan-Palay V., Jonsson G., Palay S. L. Serotonin and substance P coexist i, neurons of the rat's central nervous system. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1582–1586. doi: 10.1073/pnas.75.3.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cuello A. C., Galfre G., Milstein C. Detection of substance P in the central nervous system by a monoclonal antibody. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3532–3536. doi: 10.1073/pnas.76.7.3532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cuello A. C., Milstein C., Priestley J. V. Use of monoclonal antibodies in immunocytochemistry with special reference to the central nervous system. Brain Res Bull. 1980 Sep-Oct;5(5):575–587. doi: 10.1016/0361-9230(80)90264-6. [DOI] [PubMed] [Google Scholar]
  5. Henry J. L. Effects of substance P on functionally identified units in cat spinal cord. Brain Res. 1976 Sep 24;114(3):439–451. doi: 10.1016/0006-8993(76)90965-3. [DOI] [PubMed] [Google Scholar]
  6. Hunt S. P., Kelly J. S., Emson P. C. The electron microscopic localization of methionine-enkephalin within the superficial layers (I and II) of the spinal cord. Neuroscience. 1980;5(11):1871–1890. doi: 10.1016/0306-4522(80)90036-6. [DOI] [PubMed] [Google Scholar]
  7. Hökfelt T., Johansson O., Ljungdahl A., Lundberg J. M., Schultzberg M. Peptidergic neurones. Nature. 1980 Apr 10;284(5756):515–521. doi: 10.1038/284515a0. [DOI] [PubMed] [Google Scholar]
  8. Hökfelt T., Ljungdahl A., Steinbusch H., Verhofstad A., Nilsson G., Brodin E., Pernow B., Goldstein M. Immunohistochemical evidence of substance P-like immunoreactivity in some 5-hydroxytryptamine-containing neurons in the rat central nervous system. Neuroscience. 1978;3(6):517–538. doi: 10.1016/0306-4522(78)90017-9. [DOI] [PubMed] [Google Scholar]
  9. Jessell T. M., Iversen L. L. Opiate analgesics inhibit substance P release from rat trigeminal nucleus. Nature. 1977 Aug 11;268(5620):549–551. doi: 10.1038/268549a0. [DOI] [PubMed] [Google Scholar]
  10. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  11. Ljungdahl A., Hökfelt T., Nilsson G. Distribution of substance P-like immunoreactivity in the central nervous system of the rat--I. Cell bodies and nerve terminals. Neuroscience. 1978;3(10):861–943. doi: 10.1016/0306-4522(78)90116-1. [DOI] [PubMed] [Google Scholar]
  12. Miller R. J., Chang K. J., Cooper B., Cuatrecasas P. Radioimmunoassay and characterization of enkephalins in rat tissues. J Biol Chem. 1978 Jan 25;253(2):531–538. [PubMed] [Google Scholar]
  13. Preston R. J., McCrea R. A., Chang H. T., Kitai S. T. Anatomy and physiology of substantia nigra and retrorubral neurons studied by extra- and intracellular recording and by horseradish peroxidase labeling. Neuroscience. 1981;6(3):331–344. doi: 10.1016/0306-4522(81)90127-5. [DOI] [PubMed] [Google Scholar]

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