Skip to main content

Hybridization histochemistry

Experientia volume 43, pages 741–750 (1987)Cite this article

Summary

The location of gene expression by hybridization histochemistry is being applied in many areas of research and diagnosis. The aim of this technique is to detect specific mRNA in cells and tissues by hybridization with a complementary DNA or RNA probe. Requirements for optimal specificity, sensitivity, resolution and speed of detection may not all be encompassed in one simple technique suitable for all applications, thus appropriate procedures should be selected for specific objectives. With reference to published procedures and our own extensive experience, we have evaluated fixatives, probes, labels and other aspects of the technique critical to the preservation and hybridization in situ of mRNA and detection and quantition of hybrids.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Literature

  1. Ada, G. L., Humphrey, J. H., Askonas, B. A., McDevitt, H. O., and Nossal, G. J. V., Correlation of grain counts with radioactivity (125I and tritium) in autoradiography. Exp. Cell Res.41 (1966) 557–572.

    CAS  PubMed  Article  Google Scholar 

  2. Adams, S. P., Kavka, K. S., Wykes, E. J., Holder, S. B., and Galluppi, G. R., Hindered dialkylamino nucleoside phosphite reagents in the synthesis of two DNA 51mers. J. Am. Chem. Soc.105, (1983) 661–663.

    CAS  Article  Google Scholar 

  3. Adams, P. S., Noonan, D., and Jeffery, W. R., A model for the organization of the poly (A) protein complex in messenger ribonucleoprotein. FEBS Lett.114 (1980) 115–118.

    CAS  PubMed  Article  Google Scholar 

  4. Agrawal, S., Christodoulu, C., and Gait, M. J., Efficient methods for attaching non-radioactive labels to the 5′ ends of synthetic oligodeoxyribonucleotides. Nucl. Acids Res.14 (1986) 6226–6245.

    Article  Google Scholar 

  5. Amara, S. G., Arriza, J. L., Leff, S. E., Swanson, L. W., Evans, R. M., and Rosenfeld, M. G., Expression in brain of a messenger RNA encoding a novel neuropeptide homologous to calcitonin generelated peptide. Science229 (1985) 1094–1097.

    CAS  PubMed  Article  Google Scholar 

  6. Ambros, P. F., Matzke, A. J. M., and Matzke, M. A., Localization of agrobacterium rhizogenes T-DNA in plant chromosomes by in-situ hybridization. EMBO5 (1986) 2073–2077.

    CAS  Google Scholar 

  7. Anderson, M. A., Cornish, E. C., Mau, S.-L., Williams, E. G., Hoggart, R., Atkinson, A., Bonig, I., Grego, B., Simpson, R., Roche, P. J., Haley, J. D., Penschow, J. D., Niall, H. D., Tregear, G. W., Coghlan, J. P., Crawford, R. J., and Clarke, A. E., Cloning of cDNA for a stylar glycoprotein associated with expression of self-incompatibility inNicotiniana alata. Nature321 (1986) 38–44.

    CAS  Article  Google Scholar 

  8. Angerer, L. M., and Angerer, R. C., Detection of poly (A)+ RNA in sea urchin eggs and embryos by quantitative in-situ hybridization. Nucl. Acids Res.9 (1981) 2819–2840.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  9. Arentzen, R., Baldino, F. Jr, Davis, L. G., Higgins, G. A., Lin, Y., Manning, W., and Wolfson, B., In-situ hybridization of putative somatostatin mRNA within hypothalamus of the rat using synthetic oligonucleotide probes. J. Cell Biochem27 (1985) 415–422.

    CAS  PubMed  Article  Google Scholar 

  10. Bauman, J. G. J., Wiegant, J., Borst, P., and von Duijn, P., A new method of fluorescence microscopical localisation of specific DNA sequences by in-situ hybridization of fluorechrome labelled RNA. Exp. Cell Res.128 (1980) 485–490.

    CAS  PubMed  Article  Google Scholar 

  11. Bauman, J. G. J., Wiegant, J., and van Duijn, P., Cytochemical hybridization with fluorochrome-labelled RNA II. Applications. J. Histochem Cytochem.29 (1981) 238–246.

    CAS  PubMed  Article  Google Scholar 

  12. Bauman, J. G. J., Wiegant, J., and van Duijn, P., Cytochemical hybridization with fluorochrome-labelled RNA III. Increased sensitivity by the use of anti-fluorescein antibodies. Histochemistry73 (1981) 181–185.

    CAS  PubMed  Article  Google Scholar 

  13. Beaucage, S. L., and Caruthers, M. H., Deoxynucleoside phosphoramidites — A new class of key intermediates for deoxypolynucleotide synthesis. Tetrahedron Lett.22 (1981) 1859–1862.

    CAS  Article  Google Scholar 

  14. Blum, H. E., Stowring, L., Figus, A., Mongomery, C. K., Haase, A., and Vyas, G. N., Detection of hepatitis B virus DNA in hepatocytes, bile duct epithelium, and vascular elements by in-situ hybridization. Proc. natl Acad. Sci. USA80 (1983) 6685–6687.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  15. Bloch, B., LeGuellec, D., and de Keyzer, Y., Detection of the messenger RNAs coding for the opioid peptide precursors in pituitary and adrenal by in situ hybridization: study in several mammalian species. Neurosci. Lett.53 (1985) 141–148.

    CAS  PubMed  Article  Google Scholar 

  16. Bloch, B., Milner, R. J., Baird, A., Gubler, E., Reymond, C., Bohlen, P., le Guellec, D., and Bloom, F. E., Detection of the messenger RNA coding for preproenkephalin A in bovine adrenal in situ hybridization. Reg. Peptides8 (1984) 345–354.

    CAS  Article  Google Scholar 

  17. Brahic, M., and Haase, A. T., Detection of viral sequences of low reiteration frequency by in-situ hybridization. Proc. natl Acad. Sci. USA75 (1978) 6125–6129.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  18. Brahic, M., Haase, A. T., and Cash, E., Simultaneous in-situ detection of viral RNA and proteins. Proc. natl Acad. Sci. USA81 (1984) 5445–5448.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  19. Brigati, D. J., Myerson, D., Leary, J. J., Spalholz, B., Travis, S. Z., Fong, C. K. Y., Hsiung, G. D., and Ward, D. C., Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labelled hybridization probes. Virology126 (1983) 32–50.

    CAS  PubMed  Article  Google Scholar 

  20. Buongiorno-Nardelli, S., and Amaldi, F., Autoradiographic detection of mulecular hybrids between RNA and DNA in tissue sections. Nature225 (1970) 946–948.

    CAS  PubMed  Article  Google Scholar 

  21. Coghlan, J. P., Aldred, P., Butkus, A., Crawford, R. J., Darby, J. A., Fernley, R. T., Haralambidis, J., Hudson, P. J., Mitri, R., Niall, H. D., Penschow, J. D., Roche, R. J., Scanlon, D. B., Tregear, G. W., Richards, R., van Leeuwen, B. L., Rall, L., Scott, J., and Bell, G., Hybridization histochemistry, in: Endocrinology, pp. 18–24. Eds R. Labrie and L. Proulx. Excerpta Medica, Amsterdam 1984.

    Google Scholar 

  22. Coghlan, J. P., Penschow, J. D., Tregear, G. W., and Niall, H. D., Hybridization histochemistry: Use of complementary DNA for tissue localization of specific mRNA populations, in: Receptors, Membranes and Transport Mechanisms in Medicine, pp. 1–11. Eds A. Doyle and F. Medelson. Excerpta Medica, Amsterdam 1984.

    Google Scholar 

  23. Coghlan, J. P., Penschow, J. D., Hudson, P. J., and Niall, H. D., Hybridization histochemistry: Use of recombinant DNA for tissue localizations of specific mRNA populations. Clin. exp. Hypert.A6 (1984) 63–78.

    CAS  Google Scholar 

  24. Coghlan, J. P., Aldred, P., Haralambidis, J., Niall, H. D., Penschow, J. D., and Tregear, G. W., Hybridization histochemistry. Analyt. Biochem.149 (1985) 1–28.

    CAS  PubMed  Article  Google Scholar 

  25. Coghlan, J. P., Aldred, P., Butkus, A., Crawford, R. J., Darby, I. A., Haralambidis, J., Penschow, J. D., Roche, P. J., Troiani, C., and Tregear, G. W., Hybridization histochemistry — locating gene expression, in: Neuroendocrine Molecular Biology, pp. 33–39. Eds G. Fink, A. J. Harmar and K. W. McKerns. Plenum Publ. Corp., 1986.

  26. Coghlan, J. P., Penschow, J. D., Fraser, J. R., Aldred, P., Haralambidis, J., and Tregear, G. W., Location of gene expression in mammalian cells, in: In-Situ Hybridization. Eds K. Valentino, J. Eberwine and J. D. Barchas. Cambridge Press 1987, in press.

  27. Cox, K. H., DeLeon, D. V., Angerer, L. M., and Angerer, R. C., Detection of mRNA's in sea urchin embryos by in-situ hybridization using asymmetric RNA probes. Devl Biol.101 (1984) 485–502.

    CAS  Article  Google Scholar 

  28. Craig, R. K., Hall, L., Edbrooke, M. R., Allison, J., and MacIntyre, I., Partial nucleotide sequence of human calcitonin precursor mRNA identifies flanking cryptic peptides. Nature295 (1982) 345–347.

    CAS  PubMed  Article  Google Scholar 

  29. Dalgarno, L., Rice, C. M., and Strauss, J. H., Ross River Virus 26S RNA: complete nucleotide sequence and deduced sequence of the encoded structural proteins. Virology129 (1983) 170–187.

    CAS  PubMed  Article  Google Scholar 

  30. Darby, I. A., Aldred, G. P., Coghlan, J. P., Fernley, R. T., Penschow, J. D., and Ryan, G. B., Use of synthetic oligonucleotide and recombinant DNA probes to study renin gene expression. Clin. exp. Pharm. Physiol.12 (1985) 199–203.

    CAS  Article  Google Scholar 

  31. Darby, I. A., Aldred, P., Crawford, R. J., Fernley, R. T., Niall, H. D., Penschow, J. D., Ryan, G. B., and Coghlan, J. P., Renin gene expression in vessels of the ovine renal cortex. J. Hypert.3 (1985) 9–11.

    CAS  Article  Google Scholar 

  32. Denhart, D. T., A membrane-filter technique for the detection of complementary DNA. Biochem. biophys. Res. Commun.23 (1966) 641–646.

    Article  Google Scholar 

  33. Deschepper, C. F., Mellon, S. H., Cumin, F., Baxter, J. D., and Ganong, W. F., Analysis by immunocytochemistry and in-situ hybridization of renin and its mRNA in kidney, testis, adrenal and pituitary of rat. Proc. natl. Acad. Sci. USA83 (1986) 7552–7556.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  34. Edwards, M. K., and Wood, W. B., Location of specific messenger RNA's inCaenorhabditis elegans by cytological hybridization. Devl Biol.97 (1983) 375–390.

    CAS  Article  Google Scholar 

  35. Favre, A., Morel, C., and Scherrer, K., The secondary structure and poly (A) content of globin messenger RNA as a pure RNA and in polyribosome-derived ribonucleoprotein complexes. Eur. J. Biochem.57 (1975) 147–157.

    CAS  PubMed  Article  Google Scholar 

  36. Forster, A. C., McInnes, J. L., Skingle, D. C., and Symons, R. H., Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin. Nucl. Acids Res.13 (1985) 746–761.

    Article  Google Scholar 

  37. Fuller, P. J., Clements, J. A., Whitfield, P. L., and Funder, J. W., Kallikrein gene expression in the rat anterior pituitary. Molec. cell. Endocr.39 (1985) 99–105.

    CAS  PubMed  Article  Google Scholar 

  38. Gall, J., and Pardue, M., Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc. natl. Acad. Sci. USA63 (1969) 378–383.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  39. Galloway, D. A., Fenoglio, C. M., Shevchuk, M., and McDougall, J. K., Detection ofHerpes simplex RNA in human sensory ganglia. Virology95 (1979) 265–268.

    CAS  PubMed  Article  Google Scholar 

  40. Gee, C. E., Chen, C.-L. C., and Roberts, J. L., Identification of proopiomelanocortin neurones in rat hypothalamus by in-situ cDNA-mRNA hybridization. Nature306 (1983) 374–376.

    CAS  PubMed  Article  Google Scholar 

  41. Gee, C. E., and Roberts, J. L., In-situ hybridization histochemistry: a technique for the study of gene expression in single cells. DNA2 (1983) 155–161.

    Article  Google Scholar 

  42. Gerhard, D. S., Kawasaki, E. S., Carter Bancroft, F., and Szabo, P., Localization of a unique gene by direct hybridization in-situ. Proc. natl. Acad. Sci. USA78 (1981) 3755–3759.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  43. Gillespie, D., and Spiegelman, S., A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. molec. Biol.12 (1965) 829–842.

    CAS  PubMed  Article  Google Scholar 

  44. Godard, C. M., and Jones, K. W., Improved method for detection of cellular transcripts by in-situ hybridization. Detection of poly (A) sequences in individual cells. Histochemistry65 (1980) 291–300.

    CAS  PubMed  Article  Google Scholar 

  45. Goochee, C., Rasband, W., and Sokoloff, L., Computerized densitometry and colour coding of [14C] deoxyglocuse autoradiographs. Ann. Neurol.7 (1980) 359–370.

    CAS  PubMed  Article  Google Scholar 

  46. Gowans, E. J., Burrell, C. J., Jilbert, A. R., and Marmion, B. P., Detection of hepatitis B virus sequences in infected hepatocytes by in-situ cytohybridization. J. med. Virol.8 (1981) 67–78.

    CAS  PubMed  Article  Google Scholar 

  47. Haase, A. T., Gantz, G., Eble, B., Walker, D., Stowring, L., Ventura, P., Blum, H., Wietgrefe, S., Zupancic, M., Tourtellotte, W., Gibbs, C. J. Jr, Norrby, E., and Rozenblatt, S., Natural history of restricted synthesis and expression of measles virus genes in subacute sclerosing panencephalitis. Proc. natl. Acad. Sci. USA82 (1985) 3020–3024.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  48. Haase, A. T., Walker, D., Stowring, L., Ventura, P., Geballe, A., Blum, H., Brahic, M., Goldberg, R., and O'Brien, K., Detection of two viral genomes in single cells by double label hybridization in-situ and colour microradioautography. Science227 (1985) 189–191.

    CAS  PubMed  Article  Google Scholar 

  49. Hafen, E., Levine, M., Garber, R. L., and Gehring, W. J., An improved in-situ hybridization method for the detection of cellular RNA's inDrosophila tissue sections and its application for localising transcripts of the homeotic Antennapedia gene complex. EMBO2 (1983) 617–623.

    CAS  Google Scholar 

  50. Hall, B. D., and Spiegelman, S., Sequence complementarity of T-2 DNA and T-2 specific RNA. Proc. natl. Acad. Sci. USA47 (1961) 137–163.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  51. Harper, M. E., Marselle, L. M., Gallo, R. C., and Wong-Staal, F., Detection of lymphocytes expressing human T-lymphotropic virus type III in lymph nodes and peripheral blood from infected individuals by in-situ hybridization. Proc. natl. Acad. Sci. USA83 (1986) 772–776.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  52. Hayashi, S., Gillam, I. C., Delaney, A. D., and Tener, G. M., Acetylation of chromosome squashes ofDrosophila melanogaster decreases the background in autoradiographs from hybridization with125I-labelled RNA. J. Histochem. Cytochem.36 (1978) 677–679.

    Article  Google Scholar 

  53. Hopman, A. H. N., Wiegant, J., Tesser, G. I., and Van Duijn, P., A nonradioactive in-situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. Nucl. Acids Res.14 (1986) 6471–6488.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  54. Hudson, P., Penschow, J., Shine, J., Ryan, G., Niall, H., and Coghlan, J., Hybridization histochemistry: Use of recombinant DNA as a ‘homing probe’ for tissue localisation of specific mRNA populations. Endocrinology108 (1981) 353–356.

    CAS  PubMed  Article  Google Scholar 

  55. Hutchinson, N. J., Langer-Sofer, P. R., Ward, D. C., and Hamkalo, B. A., In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold. J. Cell Biol.95 (1982) 609–618.

    Article  Google Scholar 

  56. Jablonski, E., Moomaw, E. W., Tullis, R. H., and Ruth, J. L., Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes. Nucl. Acids Res.14 (1986) 6114–6128.

    Article  Google Scholar 

  57. Jacobs, J., Todd, K., Birnstiel, M. L., and Bird, A., Molecular hybridization of3H-labelled ribosomal RNA with DNA in ultrathin sections prepared for electron microscopy. Biochim. biophys. Acta228 (1971) 761–766.

    Article  Google Scholar 

  58. Jacobs, J. W., Simpson, E., Penschow, J., Hudson, P., Coghlan, J., and Niall, H., Characterisation and localisation of calcitonin messenger ribonucleic acid in rat thyroid. Endocrinology113 (1983) 1616–1622.

    CAS  PubMed  Article  Google Scholar 

  59. Jamrich, M., Mahon, K. A., Gavis, E. R., and Gall, J. G., Histone RNA in amphibian oocytes visualized by in-situ hybridization to methacrylate-embedded tissue sections. EMBO3 (1984) 1939–1943.

    CAS  Google Scholar 

  60. John, H. A., Birnstiel, M. L., and Jones, K. W., RNA-DNA hybrids at the cytological level. Nature223 (1969) 582–587.

    CAS  PubMed  Article  Google Scholar 

  61. Jones, K. W., Bishop, J. O., and Brito-da-Cuhna, A., A complex formation between Poly-r-(U) and various chromosomal loci inRhyncosciara. Chromosoma43 (1973) 375–390.

    CAS  PubMed  Article  Google Scholar 

  62. Karnovsky, M. J., A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J. Cell Biol.27 (1965) 137A-140A.

    Google Scholar 

  63. Koenig, S., Gendelman, H. E., Orenstein, J. M., Dal Canto, M. C., Pezeshkpour, G. H., Yungbluth, M., Janotta, F., Askamit, A., Martin, M. A., and, Fauci, A. S., Detection of AIDS virus in macrophages in brain tissue from AIDS patients with encephalopathy. Science223 (1986) 1089–1093.

    Article  Google Scholar 

  64. Kornberg, T., Sidén, I., O'Farrell, P., and Simon, M., The engrailed locus ofDrosophila: in-situ localization of transcripts reveal compartment-specific expression. Cell40 (1985) 45–53.

    CAS  PubMed  Article  Google Scholar 

  65. Kretschner, P. J., Coon, H. C., Davis, A., Harrison, M., and Nienhuis, A. W., Haemoglobin switching in sheep. J. biol. Chem.256 (1981) 1975–1982.

    Google Scholar 

  66. Landegent, J. E., Jansen in de Wal, N., Ploen, J. S., and van der Ploeg, M., Sensitive detection of hybridocytochemical results by means of reflection-contrast microscopy. J. Histochem. Cytochem.33 (1985) 1241–1246.

    CAS  PubMed  Article  Google Scholar 

  67. Langer-Sofer, P. R., Levine, M., and Ward, D. C., Immunological method for mapping genes on Drosophila polytene chromosomes. Proc. natl. Acad. Sci. USA79 (1982) 4381–4385.

    Article  Google Scholar 

  68. Lawrence, J. B., and Singer, R. H., Quantitative analysis of in-situ hybridization methods for the detection of actin gene expression. Nucl. Acids Res.13 (1985) 1777–1799.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  69. Lawrence, J. B., and Singer, R. H., Intracellular localization of messenger RNA's for cytoskeletal proteins. Cell45 (1986) 407–415.

    CAS  PubMed  Article  Google Scholar 

  70. McAllister, L. B., Scheller, R. H., Kandel, E. R., and Axel, R., In-situ hybridication to study the origin and fate of identified neurones. Science222 (1983) 800–808.

    CAS  PubMed  Article  Google Scholar 

  71. McDougall, J. K., Myerson, D., and Beckmann, A., Detection of viral DNA and RNA by in-situ hybridization. J. Histochem. Cytochem.34 (1986) 33–38.

    CAS  PubMed  Article  Google Scholar 

  72. McGinnis, W., Levine, M. S., Hafen, E., Kuroiwa, A., and Gehring, W. J., A conserved DNA sequence in homoeotic genes of theDrosophila Antennapedia and bithorax complexes. Nature308 (1984) 428–433.

    CAS  PubMed  Article  Google Scholar 

  73. McWilliams, D., and Boime, I., Cytological localisation of placental lactogen messenger ribonucleic acid in syncytiotrophoblast layers of human placenta. Endocrinology107 (1980) 761–765.

    CAS  PubMed  Article  Google Scholar 

  74. Maxam, A. M., and Gilbert, W., Sequencing end-labelled DNA with base-specific chemical cleavages. Meth. Enzymol.65 (1980) 499–560.

    CAS  PubMed  Article  Google Scholar 

  75. Melton, D. A., Krieg, P. A., Rebagliati, M. R., Maniatis, T., Zinn, K., and Green, M. R., Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP-6 promoter. Nucl. Acids Res.12 (1984) 7035–7056.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  76. Mendelsohn, F. A. O., Quirion, R., Saavedra, J. M., Aguilera, G., and Catt, K. J., Autoradiographic localization of angiotensin II receptors in rat brain. Proc. natl. Acad. Sci. USA81 (1984) 1575–1579.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  77. Moench, T. R. Gendelman, H. E., Clements, J. E., Narayan, O., and Griffin, D. E., Efficiency of in-situ hybridization as a function of probe size and fixation technique. J. virol. Meth.11 (1985) 119–130.

    CAS  Article  Google Scholar 

  78. Muragagi, A., and Wallace, R. B., Biotin-labelled oligonucleotides: enzymatic synthesis and use as hybridization probes. DNA3 (1984) 269–277.

    Article  Google Scholar 

  79. Ohkubo, H., Kageyama, R., Ujihara, M., Hirose, T., Inayama, S., and Nakanishi, S., Cloning and sequence analysis of cDNA for rat angiotensinogen. Proc. natl. Acad. Sci. USA80 (1983) 2196–2200.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  80. Orth, G., Jeantur, P., and Croissant, O., Evidence for and localisation of vegetative viral DNA replication by autoradiographic detection of RNA-DNA hybrids in sections of tumours induced by Shope papilloma virus. Proc. natl. Acad. Sci. USA68 (1971) 1876–1880.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  81. Panthier, J. J., Foote, S., Chambrand, B., Strosberg, A. D., Corvol, P., and Rougeon, F., Complete amino acid sequence and maturation of the mouse submaxillary gland renin precursor. Nature298 (1982) 90–92.

    CAS  PubMed  Article  Google Scholar 

  82. Penschow, J. D., Haralambidis, J., Aldred, P., Tregear, G. W., and Coghlan, J. P., Location of gene expression in CNS using hybridization histochemistry. Meth. Enzymol.124 (1986) 534–548.

    CAS  PubMed  Article  Google Scholar 

  83. Pfeifer-Ohlsson, S., Goustin, A. S., Rydnert, J., Bjersing, L., Wahlström, T., Stehelin, D., and Ohlsson, R., Spatial and temporal pattern of cellular myc oncogene expression in developing human placenta: implications for embryonic cell proliferation. Cell38 (1984) 585–596.

    CAS  PubMed  Article  Google Scholar 

  84. Pintar, J. E., Schachter, B. S., Herman, A. B., Durgerian, S., and Krieger, D. T., Characterization and localization of proopiomelanocortin messenger RNA in the adult rat testis. Science225 (1984) 632–634.

    CAS  PubMed  Article  Google Scholar 

  85. Pochet, R., Brocas, H., Vassart, G., Toubeau, G., Seo, H., Refetoff, S., Dumont, J. E., and Pasteels, J. L., Radioisotopic localisation of prolactin mRNA on histological sections by in-situ hybridization. Brain Res.211 (1981) 433–438.

    CAS  PubMed  Article  Google Scholar 

  86. Rall, L. B., Scott, J., Bell, G. I., Crawford, R. J., Penschow, J. D., Niall, M. D., and Coghlan, J. P., Mouse prepro-epidermal growth factor synthesis by the kidney and other tissues. Nature313 (1985) 228–231.

    CAS  PubMed  Article  Google Scholar 

  87. Richards, R. I., Catanzaro, D. F., Mason, A. J., Morris, B. J., Baxter, J. D., and Shine, J., Mouse glandular kallikrein genes. J. biol. Chem.257 (1982) 2758–2761.

    CAS  PubMed  Google Scholar 

  88. Rigby, P. W. J., Dieckmann, M., Rhodes, C., and Berg, P., Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. molec Biol.113 (1977) 237–241.

    CAS  PubMed  Article  Google Scholar 

  89. Robins, D. M., Ripley, S., Henderson, A. S., and Axel, R., Transforming DNA integrates into the host chromosome. Cell23 (1981) 29–39.

    CAS  PubMed  Article  Google Scholar 

  90. Rogers, A. W., Techniques of autoradiography, pp. 201–284. Elsevier, North Holland, Amsterdam 1979.

    Google Scholar 

  91. Rosenfeld, M. G., Mermod, J.-J., Amara, S. G., Swanson, L. W., Sawchenko, P. E., Rivier, J., Vale, W. W., and Evans, R. M., Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature304 (1983) 129–135.

    CAS  PubMed  Article  Google Scholar 

  92. Rudkin, G. T., and Stollar, B. D., High resolution detection of DNA-RNA hybrids in-situ by indirect immunofluorescence. Nature265 (1977) 472–473.

    CAS  PubMed  Article  Google Scholar 

  93. Schalling, M., Hökfelt, T., Wallace, B., Goldstein, M., Filer, D., Yamin, C., and Schlesinger, D. H., Tyrosine-3-hydroxylase in rat brain and adrenal medulla: hybridization histochemistry and immunohistochemistry combined with retrograde tracing. Proc. natl. Acad. Sci. USA83 (1986) 6208–6212.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  94. Scheller, R. H., Kaldany, R.-R., Kreiner, T., Mahon, A. C., Nambu, J. R., Schaefer, M., and Taussig, R., Neuropeptides: mediators of behaviour inAplysia. Science225 (1984) 1300–1308.

    CAS  PubMed  Article  Google Scholar 

  95. Scott, J., Patterson, S., Rall, L., Bell, G. L., Crawford, R., Penschow, J. D., Niall, H. D., and Coghlan, J., The structure and biosynthesis of epidermal growth factor precursor. J. Cell Sci. Suppl.3 (1985) 19–28.

    Article  Google Scholar 

  96. Seeburg, P., The human growth hormone gene family: nucleotide sequences show recent divergence and predict a new polypeptide hormone. DNA1 (1982) 239–249.

    CAS  PubMed  Article  Google Scholar 

  97. Shivers, B. D., Harlan, R. E., Hejtmancik, J. F., Conn, P. M., and Pfaff, D. W., Localization of cells containing LHRH-like mRNA in rat forebrain using in-situ hybridization. Endocrinology118 (1986) 883–885.

    CAS  PubMed  Article  Google Scholar 

  98. Shivers, B. D., Harlan, R. E., Pfaff, D. W., and Schachter, B. S., Combination of immunocytochemistry and in-situ hybridization in the same tissue section of rat pituitary. J. Histochem. Cytochem.34 (1986) 39–43.

    CAS  PubMed  Article  Google Scholar 

  99. Shivers, B. D., Schachter, B. S., and Pfaff, D. W., In-situ hybridization for the study of gene expression in the brain. Meth. Enzymol.124 (1986) 497–510.

    CAS  PubMed  Article  Google Scholar 

  100. Singer, R. H., and Ward, D. C., Actin gene expression visualized in chicken muscle tissue culture by using in-situ hybridzation with a biotinated nucleotide analog. Proc. natl. Acad. Sci. USA79 (1982) 7331–7335.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  101. Smith, F. M., Fung, S., Hunkapiller, M. W., Hunkapiller, T. J., and Hood, L. E., The synthesis of oligonucleotides containing an aliphatic amino group at the S'terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis. Nucl. Acids Res.13 (1985) 2399–2502.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  102. Stuart, W. D., and Porter, D. L., An improved in-situ hybridization method. Exp. Cell Res.113 (1978) 219–222.

    CAS  Article  Google Scholar 

  103. Szabo, P., Elder, E., Steffensen, D. M., and Uhlenbeck, O. C., Quantitative in-situ hybridization of ribosomal RNA species to polytene chromosomes ofDrosophila melanogaster. J. molec. Biol.115 (1977) 539–563.

    CAS  PubMed  Article  Google Scholar 

  104. Taylor, J. M., Illmensee, R., and Summers, J., Efficient transcription of RNA into DNA by avian sarcoma virus polymerase. Biochim. biophys. Acta442 (1976) 324–330.

    CAS  PubMed  Article  Google Scholar 

  105. Tu, C-P. D., and Cohen, S. N., 3′end-labelling of DNA with [α-32P] cordycepin-5-triphosphate. Gene10 (1980) 177–181.

    CAS  PubMed  Article  Google Scholar 

  106. van Leeuwen, B. H., Evans, B. A., Tregear, G. W., and Richards, R. I., Mouse glandular kallikrein genes. J. biol. Chem.261 (1986) 5529–5535.

    PubMed  Google Scholar 

  107. Varndell, I. M., Polak, J. M., Sikri, K. L., Minth, C. D., Bloom, S. R., and Dixon, J. E., Visualisation of messenger RNA directing peptide synthesis by in-situ hybridization using a novel single-stranded cDNA probe. Histochemistry81 (1984) 597–601.

    CAS  PubMed  Article  Google Scholar 

  108. Venezky, D. L., Angerer, L. M., and Angerer, R. C., Accumulation of histone repeat transcripts in the sea urchin egg pronucleus. Cell24 (1981), 385–391.

    CAS  PubMed  Article  Google Scholar 

  109. Wahl, G. M., Stern, M., and Stark, G. R., Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization using dextran sulphate. Proc. natl. Acad. Sci. USA76 (1979) 3683–3687.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  110. Warembourg, M., Tranchant, O., Perret, C., Desplan, C., and Thomasset, M., In-situ detection of vitamin D-induced calcium binding protein (9-kDa CaBP) messenger RNA in rat duodenum. J. Histochem. Cytochem.34 (1986) 277–280.

    CAS  PubMed  Article  Google Scholar 

  111. Wilcox, J. N., Gee, C. E., and Roberts, J. L., In-situ cDNA: mRNA hybridization: development of a technique to measure mRNA levels in individual cells. Meth. Enzymol.124 (1986) 510–533.

    CAS  PubMed  Article  Google Scholar 

  112. Wintour, E. M., The role of the adrenal in control of foetal-developmental gene-switches in haemoglobin. Proc. XXXth Congress I.U.P.S. (1986) 121 (abstr.).

  113. Wintour, E. M., Smith, M. B., Bell, R. J., McDougall, J. G., and Cauchi, M. N., The role of fetal adrenal hormones in the switch from fetal to adult globin synthesis in sheep. J. Endocr.104 (1985) 165–170.

    CAS  PubMed  Article  Google Scholar 

  114. Zajac, J. D., Penschow, J. D., Mason, T., Tregear, G. W., Coghlan, J. P., and Martin, T. J., Identification of calcitonin and calcitonin gene-related peptide messenger ribonucleic acid in medullary thyroid carcinomas by hybridization histochemistry. J. clin. Endocr. Metab.62 (1986) 1037–1043.

    CAS  PubMed  Article  Google Scholar 

  115. Zawatsky, R., De Maeyer, E., and De Maeyer-Guignard, J., Identification of individual interferon-producing cells by in-situ hybridization. Proc. natl. Acad. Sci. USA82 (1985) 1136–1140.

    Article  Google Scholar 

Download references

Author information

Affiliations

  1. Howard Florey Institute of Experimental Physiology and Medicine, University of Melbourne, 3052, Parkville, Victoria, Australia

    J. D. Penschow, J. Haralambidis, P. E. Darling, I. A. Darby, E. M. Wintour, G. W. Tregear & J. P. coghlan

Authors
  1. J. D. Penschow
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Haralambidis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. E. Darling
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I. A. Darby
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E. M. Wintour
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. G. W. Tregear
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J. P. coghlan
    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

Penschow, J.D., Haralambidis, J., Darling, P.E. et al. Hybridization histochemistry. Experientia 43, 741–750 (1987). https://doi.org/10.1007/BF01945351

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01945351

Key words

  • Hybridization histochemistry
  • hybridocytochemistry
  • in situ hybridization
  • gene expression
  • histochemical hybridization
  • nucleic acid hybridization
  • histocytochemistry