Summary
Quantitative autoradiography of macroscopic specimens using computer-assisted image analysis is now widely used for studying the distribution of peptide receptors in the brain and peripheral tissues and more recently has been used to measure mRNA in tissue sections by in situ hybridisation. The spatial distribution of radiolabelled substances in tissue can be detected by the blackening of the emulsion in sheets of radiation-sensitive film and the resulting pattern of optical densities within the autoradiogram can be quantified by comparison with a calibrated radioactive scale. In this review, the technique of computer-assisted densitometry is described together with guidelines for the selection and preparation of radioactive standards. Strategies are discussed for ensuring that radioactivity can be measured and quantified using film-based emulsions with a precision approaching that of conventional counting techniques.
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References
Baskin, D. G., and Dorsa D. M., Quantitative autoradiography and in vitro radioligand binding, in: Functional Mapping in Biology and Medicine: Computer-assisted Autoradiography, pp. 204–234. Ed. D. L. McEachron. Karger, Basel 1986.
Baskin, D. G., Davidson, D., Corp, E. S., Lewellen, T., and Graham, M., An inexpensive microcomputer digital imaging system for densitometry: quantitative autoradiography of insulin receptors with [125I] and LKB Ultrofilm. J. Neurosci. Meth. 16 (1986) 119–129.
Baldino, F., and Davis, L. G., Gluccocorticoid regulation of vasopressin messenger RNA, in: In Situ Hybridization in Brain, pp. 97–116. Ed. G. R. Uhl. Plenum Press, New York 1986.
Beresford, I. J. M., Davenport, A. P., Hall, M. D., Hill, R. G., and Hughes, J., Autoradiographic localization of CCK receptors on intrinsic striatal neurones. Neuroscience 22 (1987a) S803.
Beresford, I. J. M., Hall, M. D., Clark, C. R., Hill, R. G., Hughes, J., and Sirinathsinghji, D. J. S., Striatal lesions and transplants demonstrate that cholecystokinin receptors are localized on intrinsic striatal neurones: a quantitative autoradiographic study. Neuropeptides 10 (1987b) 109–136.
Beresford, I. J. M., Davenport, A. P., Hall, M. D., Hill, R. G., Hughes, J., and Sirinathsinghji, D. J. S., Experimental hemiparkinsonism in the rat following chronic unilateral infusion of MPP+ into the nigrostriatal dopamine pathway: II. Differential localization of dopamine and cholecystokinin receptors. Neuroscience 27 (1988) in press.
Boast, C. A., Snowhill, E. W., and Altar, C. A., Quantitative Receptor Autoradiography. Alan R. Liss, New York 1986.
Brown, M. J., Davenport, A. P., Emson, P. C., Hall, M. D., and Nunez, D. J., Adrenal cells express atrial natriuretic mRNA (ANP mRNA). An ‘in-situ’ hybridization study. J. Physiol. 398 (1988a) 76P.
Brown, M. J., Davenport, A. P., Emson, P. C., Hall, M. D., and Nunez, D. J., Measurement of atrial natriuretic peptide messenger RNA (ANP mRNA) in the neonatal rat heart by computer-assisted image analysis of ‘in-situ’ hybridization (ISH). J. Physiol. 398 (1988b) 75P.
Camus, A., Javoy-agid, F., Dubois, A., and Scatton, B., Autoradiographic localization and quantification of Dopamine D2 receptors in normal human brain with [3H]N-n-propylnorapomorphine. Brain Res. 575 (1986) 135–149.
Clark, C. R., and Hall, M. D., Hormone receptor autoradiography: Recent developments. TIBS 11 (1986) 195–199.
Davenport, A. P., Hall, M. D., Hill, R. G., and Hughes, J., Evaluation of [125I] reference sources for quantitative receptor autoradiography: comparison between brain paste and polymer standards. Neuroscience 22 (1987) S371.
Davenport, A. P., and Hall, M. D., Comparison between brain paste and polymer [I25I] standards for quantitative receptor autoradiography. J. Neurosci. Meth. 25 (1988) 75–82.
Ehn, E., and Larsson, B., Properties of an antiscratch-layer-free X-ray film for the autoradiographic registration of tritium. Sci. Tools 26 (1979) 24–29.
Eilbert, J. L., Quantitative analysis of autoradiographs, in Functional Mapping in Biology and Medicine: Computer assisted Autoradiography, pp. 122–142. Ed. D. L. McEachron. Karger, Basel 1986.
Geary, W. A., and Wooten, G. F., Regional tritium quenching in quantitative autoradiography in the central nervous system. Brain Res. 336 (1985) 334–336.
Geary, W. A., Toga, A. W., and Wooten, G. F., Quantitative film autoradiography for tritium: Methodological considerations. Brain Res. 337 (1985) 99–108.
Hall, M. D., Davenport, A. P., and Clark, C. R., Quantitative receptor autoradiography. Nature 324 (1986) 493–494.
Herkenham, M., and Sokoloff, L., Quantitative receptor autoradiography: Tissue defatting eliminates differential self-absorption of tritium radiation in grey and white matter of brain. Brain Res. 321 (1984) 363–368.
Horrocks, D. L., Standardizing 125I sources and determining 125I counting efficiencies of well-type gamma counting systems. Clin. Chem. 21 (1975) 370–375.
Kuhar, M. J., Autoradiographic localization of drugs and neurotransmitter receptors, in: Handbook of Chemical Neuroanatomy: Methods in Chemical Neuroanatomy, vol. 1, pp. 398–415. A. Eds A. Bjorklund and T. Hökfelt. Elsevier, Amsterdam.
Kuhar, M. J., Recent progress in receptor mapping: which neurons contain the receptors. TINS 10 (1987) 308–310.
Kuhar, M. J., and Unnerstall, J. R., Quantitative receptor mapping by autoradiography: Some current technical problems. TINS 8 (1985) 49–53.
Kuhar, M. J., De Souza, E. B., and Unnerstall, J. R., Neurotransmitter receptor mapping by autoradiography and other methods. A. Rev. Neurosci. 9 (1986) 27–59.
Larsson, B., and Ullberg, S., A film for the registration of tritium. Sci. Tools, Special Issue on Whole Body Autoradiography (1977) 30–33.
Laskey, R. A., Radiosotope detection by fluorography and intensifying screens. Amersham International plc, Amersham, UK 1984.
Lewis, M., Pert, A., Pert, C., and Herkenham, M., Opiate receptor localization in rat cerebral cortex. J. comp. Neurol. 216 (1983) 339–358.
McEachron, D. L., Adler, N. T., and Tretiak, O. J., Two views of functional mapping and autoradiography, in: Functional Mapping in Biology and Medicine: Computer assisted Autoradiography, pp. 1–46. Ed. D. L. McEachron. Karger, Basel 1986.
Nunez, D. J., Davenport, A. P., Emson, P. C., Hall, M. D., and Brown, M. J., In-situ hybridization to localize artrial natriuetic peptide messenger RNA (ANP mRNA). Quantitation using computer-assisted (Quantimet 920) image analysis. Clin. Sci. molec. Med. Suppl. 17 (1987) 1p.
Pan, H. S., Frey, K. A., Young, A. B., and Penney, J. B., Changes in [3H]-muscimol binding in the substantia nigra, endopeduncular nucleus, globus pallidus and thalamus after striatal lesions as demonstrated by quantitative receptor autoradiography. J. Neurosci. 3 (1983) 1189–1198.
Paxinos, G., and Watson, C., The rat brain in stereotaxic coordinates. Academic Press, Sydney 1986.
Palacios, J. M., Probst, A., and Cortes R., Mapping receptors in human brain. TINS 9 (1986) 284–289.
Rainbow, T. C., Bleisch, W. V., Biegon, A., and McEwen, B. S., Quantitative densitometry of neurotransmiter receptors. J. Neuorsci. Meth. 5 (1982) 127–138.
Rainbow, T. C., Biegon, A., and Berck, D. J., Quantitative receptor autoradiography with tritium-labelled ligands: comparison of biochemical and densitometric measurements. J. Neurosci. Meth. 11 (1984) 231–241.
Rogers, A. W., Techniques of Autoradiography, 3rd edn. Elsevier, Amsterdam 1979.
Savasta, M., Dubois, A., and Scatton, B., Autoradiographic localization of DI Dopamine receptors in rat brain with [3H]SCH 23390. Brain Res. 375 (1986) 291–301.
Sherman, T. G., Kelsey, J. E., Khachaturian, H., Burke, S., Akil, H., and Watson, S. J., Opioid peptides and vasopressin: Application of in situ hybridization to studies of the hypothalamus and pituitary, in: In Situ Hybridization in Brain, pp. 49–62. Ed. G. R. Uhl. Plenum Press, New York 1986.
Shivers, B. D., Harlan, R. E., Romano, G. J., Howells, R. D., and Pfaff, D. W., Cellular location and regulation of proenkephalin mRNA in rat brain, in: In Situ Hybridization in Brain, pp. 3–20. Ed. G. R. Uhl. Plenum Press, New York 1986.
Thiel, E., Dormer, P., and Ruppelt, W., Thierfelder, S., Quantitative immunoautoradiography at the cellular level II. Absolute measurements using labelled standard cells as a source of reference. J. Immun. Meth. 12 (1986) 237–251.
Uhl, G. R., Evans, J., Parta, M., Walworth, C., Hill, K., Sasek, C., Voigt, M., and Reppert, S., Vasopressin and somatostatin mRNA in situ hybridization, in: In Situ Hybridization in Brain, pp. 21–47. Ed. G. R. Uhl. Plenum Press, New York 1986.
Ullberg, S. A., The technique of whole body autoradiography. Sci. Tools, Special Issue on Whole Body Autoradiography (1987) 2–29.
Unnerstall, J. R., Niehoff, D. L., Kuhar, M. J., and Palacios, J. M., Quantitative receptor autoradiography using [3H]Ultrofilm: application to multiple benzodiazepine receptors, J. Neurosci. Meth. 6 (1982) 59–73.
Young, W. S., and Kuhar, M. J., A new method for receptor autoradiography: [3H]opioid receptors in rat brain. Brain Res. 179 (1979) 255–270.
Young, W. S., and Kuhar, M. J., Quantitative in situ hybridization and determination of mRNA content, in: In Situ Hybridization in Brain, pp. 245–248. G. R. Uhl. Plenum Press, New York 1986.
Zilles, K., Schleicher, A., Rath, M., Glaser, T., and Traber, J., Quantitative autoradiography of transmitter binding sites with an image analyzer. J. Neurosci. Meth. 18 (1986) 207–220.
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Davenport, A.P., Hill, R.G., Hughes, J. (1989). Quantitative analysis of autoradiograms. In: Polak, J.M. (eds) Regulatory Peptides. Experientia Supplementum, vol 56. Birkhäuser Basel. https://doi.org/10.1007/978-3-0348-9136-3_8
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DOI: https://doi.org/10.1007/978-3-0348-9136-3_8
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Print ISBN: 978-3-7643-1976-2
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