Skip to main content
SpringerLink
Log in

A new method for quenching correction leads to revisions of data in receptor autoradiography

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

Differential quenching of β-emission affects strongly the analysis of receptor distribution patterns in quantitative receptor autoradiography with tritiated ligands. Different methods for the quenching correction have been described in the past, but some of these are of limited value, if a detailed anatomical parcellation is necessary. Other methods correct exclusively local variations in lipid concentration, which is an important, but only one of several factors causing quenching. A new method for the measurement of quenching (or autoradiographic efficiency) is presented, which permits an anatomically detailed and direct determination of the total quenching without lipid extraction procedures. This method is based on the measurement of autoradiographic efficiency in cryostat sections homogeneously labeled with tritiated formaldehyde by an underlying gelatine section containing this labeled compound. Regional and layer specific measurements of autoradiographic efficiency in cortical and subcortical regions of the human and rat brain are reported. A significant correlation was found between the density of myelin and autoradiographic efficiency but other factors were also shown to influence differential quenching. The use of the here presented correction procedure leads to revisions of the laminar distribution patterns reported for different receptors in human and rat cortical areas. Our results show, that a complete quenching correction is necessary for the mapping of receptor distributions with tritiated ligands.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alexander GM, Schwartzman RJ (1984) Quantitative computer analysis of autoradiographs utilizing a charge-coupled device solid-state camera. J Neurosci Methods 12:29–36

    Google Scholar 

  • Alexander GM, Schwartzman RJ, Bell RD, Renthal A (1981) Quantitative measurement of local cerebral metabolic rate for glucose utilizating tritiated 2-deoxyglucose. Brain Research 223:59–67

    Google Scholar 

  • Altar CA, Walter RJ, Neve KA (1984) Computer-assisted video analysis of [3H] spiroperidol binding autoradiographs. J Neurosci Methods 10:173–188

    Google Scholar 

  • Baskin DG, Filuk PE, Stahl WL (1989) Standardization of tritium-sensitive film for quantitative autoradiography. J Histochem Cytochem 37:1337–1344

    Google Scholar 

  • Benfenati F, Cimino M, Agnati LF, Fuxe K (1986) Quantitative autoradiography of central neurotransmitter receptors: methododical and statistical aspects with special reference to computer-assisted image analysis. Acta Physiol Scand 128:129–146

    Google Scholar 

  • Boast CA, Snowhill EW, Altar CA (1986) Quantitative receptor autoradiography. Liss, New York, pp 1–268

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochemistry 72:248–254

    Google Scholar 

  • Brodmann K (1909) Verlgeichende Lokalisationslehre der Großhirnrinde, Barth, Leipzig, pp 1–324

    Google Scholar 

  • Clark CR, Hall MD (1986) Hormone receptor autoradiography: recent developments. Trends Biochem Sci 11:195–199

    Google Scholar 

  • Davenport AP, Beresford IJM, Hall MD, Hill RG, Hughes J (1988) Quantitative autoradiography in neuroscience. In: Leeuwen FW van, Buijs RM, Pool CW, Pach O (eds) Molecular neuroanatomy. Elsevier, Amsterdam New York Oxford, pp 121–145

    Google Scholar 

  • De Lima AD, Bloom FE, Morrison JH (1988) Synaptic organizaton of serotonin-immunoreactive fibers in primary visual cortex of the macaque monkey. J Comp Neurol 274:280–294

    Google Scholar 

  • Ernsberger P, Arango V, Reis DJ (1988a) A high density of muscarinic receptors in the rostral ventrolateral medulla of the rat is revealed by correction for autoradiographic efficiency. Neurosci Lett 85:179–186

    Google Scholar 

  • Ernsberger P, Arneric SP, Arango V, Reis DJ (1988b) Quantitative distribution of muscarinic receptors and choline acetyltransferase in rat medulla: examination of transmitter-receptor mismatch. Brain Res 452:336–344

    Google Scholar 

  • Foote RW (1987) Fiber tracts that contain more opioid binding sites than gray matter does: a quantitative autoradiographic study in the guinea-pig. Neuroscience 20:109–116

    Google Scholar 

  • Gallyas F (1979) Silver staining of myelin by means of physical development. Neurol Res 1:203–209

    Google Scholar 

  • Geary WA, Wooten GF (1985) Regional tritium quenching in quantitative autoradiography of the central nervous system. Brain Res 336:334–336

    Google Scholar 

  • Geary WA, Toga AW, Wooten GF (1985) Quantitative film autoradiography for tritium: methodological considerations. Brain Res 337:99–108

    Google Scholar 

  • Hall MD, Davenport AP, Clark CR (1986) Quantitative receptor autoradiography. Nature 324:493–494

    Google Scholar 

  • Herkenham M (1985) Autoradiographic methods for receptor localization. In: Karten HJ (ed) Modern neuroanatomical methods. Neuroscience short course No. 1 syllabus. Soc Neurosci, Bethesda, Maryland, pp 44–60

    Google Scholar 

  • Herkenham M (1988a) Influence of tissue treatment on quantitative receptor autoradiography. In: Leeuwen FW van, Buijs RM, Pool CW, Pach O (eds) Molecular neuroanatomy. Elsevier, Amsterdam New York Oxford, pp 111–120

    Google Scholar 

  • Herkenham M (1988b) Mismatches between receptor and transmitter localizations in the brain. In: Leeuwen FW van, Buijs RM, Pool CW, Pach O (eds) Molecular neuroanatomy. Elsevier, Amsterdam New York Oxford, pp 219–229

    Google Scholar 

  • Herkenham M, Pert CB (1982) Light microscopic localization of brain opiate receptors: a general autoradiographic method which preserves tissue quality. J Neurosci 2:1129–1149

    Google Scholar 

  • Herkenham M, Sokoloff L (1984) Quantitative receptor autoradiography: tissue defatting eliminates differential self-absorption of tritium radiation in gray and white matter of brain. Brain Res 321:363–368

    Google Scholar 

  • Korr H (1985) Determination of correction factors of 3H-β-self-absorption for quantitative evaluation of grain number in autoradiographic studies. Interferometric studies of different cell types in the mouse brain. Histochemistry 83:65–70

    Google Scholar 

  • Kuhar MJ, Unnerstall JR (1985) Quantitative receptor mapping by autoradiography: some current technical problems. Trends Neurosci 8:49–53

    Google Scholar 

  • Kuhar MJ, DeSouza EB, Unnerstall JR (1986) Neurotransmitter receptor mapping by autoradiography and other methods. Ann Rev Neurosci 9:27–59

    Google Scholar 

  • Lidow MS, Goldman-Rakic PS, Rakic P, Gallager DW (1988) Differential quenching and limits of resolution in autoradiograms of brain tissue labeled with 3H-, 125I-and 14C-compounds. Brain Res 459:105–119

    Google Scholar 

  • Lidow MS, Goldman-Rakic PS, Gallager DW, Rakic P (1989) Quantitative autoradiographic mapping of serotonin 5-HT1 and 5-HT2 receptors and uptake sites in the neocortex of the rhesus monkey. J Comp Neurol 280:27–42

    Google Scholar 

  • Maurer W, Primbsch E (1964) Größe der β-Selbstabsorption bei der 3H-Autoradiographie. Exp Cell Res 33:8–18

    Google Scholar 

  • Merker B (1983) Silver staining of cell bodies by means of physical development. J Neurosci Methods 9:235–241

    Google Scholar 

  • Palacios JM, Niehoff DL, Kuhar MJ (1981) Receptor autoradiography with tritium-sensitive film: potential for computerized densitometry. Neurosci Lett 25:101–105

    Google Scholar 

  • Palacios JM, Cortés R, Dietl MM (1988) A laboratory for the in vitro labeling of receptors in tissue sections for autoradiography. In: Leeuwen FW van, Buijs RM, Pool CW, Pach O (eds) Molecular neuroanatomy. Elsevier, Amsterdam New York Oxford, pp 95–109

    Google Scholar 

  • Ramm P, Kulick JH, Stryker MP, Frost BJ (1984) Video and scanning microdensitometer-based imaging systems in autoradiographic densitometry. J Neurosci Methods 11:89–100

    Google Scholar 

  • Rainbow TC, Biegon A, Berck DJ (1984) Quantitative receptor autoradiography with tritium-labeled ligands: comparison of biochemical and densitometric measurements. J Neurosci Methods 11:231–241

    Google Scholar 

  • Schleicher A, Zilles K (1988) The use of automated image analysis for quantitative receptor autoradiography. In: Leeuwen FW van, Buijs RM, Pool CW, Pach O (eds) Molecular neuroanatomy. Elsevier, Amsterdam New York Oxford, pp 147–157

    Google Scholar 

  • Unnerstall JR, Niehoff DL, Kuhar MJ, Palacios JM (1982) Quantitative receptor autoradiography using [3H]Ultrofilm: application to multiple benzodiazepine receptors. J Neurosci Methods 6:59–78

    Google Scholar 

  • Unnerstall JR, Marshall RD, DeSouza EB, Kuhar MJ (1984) Receptor autoradiography in brain: coping with regional differences in autoradiography efficiency with tritium. Fed Proc 43:288

    Google Scholar 

  • Zilles K (1985) The cortex of the rat. A stereotaxic atlas. Springer, Berlin Heidelberg New York Tokyo, pp 1–121

    Google Scholar 

  • Zilles K (1988) Receptor autoradiography in the hippocampus of man and rat. Adv Anat Embryol Cell Biol 111:61–80

    Google Scholar 

  • Zilles K (1989) Codistribution of transmitter receptors in the human and rat hippocampus. In: Chan-Palay V, Köhler C (eds) The hippocampus — New vistas. Liss, New York, pp 189–205

    Google Scholar 

  • Zilles A, Schleicher A, Rath M, Glaser T, Traber J (1986) Quantitative autoradiography of transmitter binding sites with an image analyzer. J Neurosci Methods 18:207–220

    Google Scholar 

  • Zilles A, Schleicher A, Rath M, Bauer A (1988) Quantitative receptor autoradiography in the human brain. Methodical aspect. Histochemistry 90:129–137

    Google Scholar 

  • Zilles K, Schröder H, Schröder U, Horváth E, Werner L, Luiten PGM, Maelicke A, Strosberg AD (1989) Distribution of cholinergic receptors in the rat and human neocortex. In: Frotscher M, Misgeld U (eds), Central cholinergic synaptic transmission. Birkhäuser, Basel Berlin Boston, pp 212–228

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute I of Anatomy, University of Cologne, Joseph-Stelzmann-Strasse 9, W-5000, Cologne 41, Federal Republic of Germany

    K. Zilles, K. zur Nieden & A. Schleicher

  2. Department of Neurobiology, Troponwerke, Berliner Strasse 156, W-5000, Cologne 80, Federal Republic of Germany

    J. Traber

Authors
  1. K. Zilles
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. zur Nieden
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Schleicher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Traber
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zilles, K., zur Nieden, K., Schleicher, A. et al. A new method for quenching correction leads to revisions of data in receptor autoradiography. Histochemistry 94, 569–578 (1990). https://doi.org/10.1007/BF00271983

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00271983

Keywords

Search

Navigation