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Use of comparative in situ hybridization and immunocytochemistry for the study of regulatory peptides

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In Situ Hybridization: Medical Applications

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Abstract

In the past decade immunocytochemistry has revolutionized the approach to morphological studies of tissues, allowing the localization of specific peptides and other cellular markers to be demonstrated. However, the knowledge gained by identifying cellular antigens is limited, as simple localization cannot give any information on the dynamic changes of cellular metabolic processes. The recent introduction of in situ hybridization for mRNA or DNA using labelled complementary nucleic acid probes has made possible the understanding of gene expression at the cellular level in different physiological and pathological conditions. Compared to other morphological and molecular biological techniques, in situ hybridization offers precise anatomical localization and increased sensitivity, opening completely new horizons to the scope of morphological studies.

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References

  • Angerer RC, Cox KH and Angerer LM (1985) In situ hybridization to cellular RNAs. Genet. Eng. 7: 43–65.

    CAS  Google Scholar 

  • Bhatt B, Burns J, Flannery D and McGee JO’D (1988) Direct visualisation of single copy genes on banded metaphase chromosome by nonisotopic in situ hybridization. Nuc. Acids Res. 16: 3951–3961.

    Article  CAS  Google Scholar 

  • Bishop AE, Hamid QA, Adams C, Bretherton-Watt D, Jones PM, Denny P, Stamp GWM, Hurt RL, Grimelius L, Harmar AJ, Valentino K, Cedermark B, Legon S, Ghatei MA, Bloom SR and Polak JM (1989) Expression of tachykinins by ileal and lung carcinoid tumours assessed by combined in situ hybridization, immunocytochemistry and radioimmunoassay. Cancer 63: 1129–1137.

    Article  PubMed  CAS  Google Scholar 

  • Brahic M, Haase AT and Cash E (1984) Simultaneous in situ detection of viral RNA and antigens. Proc. Natl. Acad. Sci. USA 81: 5445–5448.

    Article  PubMed  CAS  Google Scholar 

  • Brysch W, Hagendorff G and Schlingensiepen (1988) RNA probes transcribed from synthetic DNA for in situ hybridization. Nuc. Acids Res. 16: 2333.

    Article  CAS  Google Scholar 

  • Casey J and Davidson N (1977) Rates of formation and thermal stabilities of RNA:RNA and DNA:DNA duplexes at high concentration of formamide. Nuc. Acids Res. 4: 1539–1552.

    Article  CAS  Google Scholar 

  • Chan-Palay V, Yasargil G, Hamid Q, Polak JM and Palay SL (1988) Simultaneous demonstration of neuropeptide Y gene expression and peptide storage in single neurons of the human brain. Proc. Natl. Acad. Sci. USA 85: 3213–3215.

    Article  PubMed  CAS  Google Scholar 

  • Cox KH, De Leon DV, Angerer LM and Angerer RC (1984) Detection of mRNA’s in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101: 485–502.

    Article  PubMed  CAS  Google Scholar 

  • Davenport AP and Nunez DJ (1990) Quantification in in situ hybridization. In: In Situ Hybridization — Principles and Practice. (Polak JM and McGee JO’D eds), Oxford University Press.

    Google Scholar 

  • Denny P, Hamid Q, Krause JE, Polak JM and Legon S (1988) Oligoriboprobes: tools for in situ hybridization. Histochemistry 84: 481–483.

    Article  Google Scholar 

  • Giaid A, Hamid H, Adams C, Springall DR, Terenghi G, Polak JM (1989a) Non-isotopic RNA probes. Comparison between different labels and detection systems. Histochemistry 93: 191–196.

    CAS  Google Scholar 

  • Giaid A, Gibson SJ, Ibrahim NBN, Legon S, Bloom SR, Yanagisawa M, Masaki T, Varndell IM and Polak JM (1989b) Endothelin 1, an endothelium-derived peptide is expressed in neurons of the human spinal cord and dorsal root ganglia. Proc. Natl. Acad. Sci. USA 86: 7634–7638.

    Article  PubMed  CAS  Google Scholar 

  • Gibson SJ, Polak JM, Bloom SR, Sabate IM, Mulderry PK, Morrison JFB, Kelly JS, Rosenfeld MG and Evans R (1984) Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and eight mammalian species. J. Neurosci. 4: 3101–3111.

    PubMed  CAS  Google Scholar 

  • Gibson SJ, Polak JM, Giaid A, Hamid QA, Kar S, Jones PM, Denny P, Legon S, Amara SG, Craig RK, Bloom SR, Penketh RJA, Rodek C, Ibraim NBN and Dawson A (1988) Calcitonin gene-related peptide mRNA is expressed in sensory neurons of the dorsal root ganglia and also in spinal motoneurones in man and rat. Neurosci. Lett. 91: 283–288.

    Article  PubMed  CAS  Google Scholar 

  • Gillam IC (1987) Non-radioactive probes for specific DNA sequences. Tib. Tech. 5: 332–334.

    CAS  Google Scholar 

  • Hamid Q, Wharton J, Terenghi G, Hassall CJ, Aimi J, Taylor KM, Nakazato H, Dixon JE, Burnstock G and Polak JM (1987) Localization of atrial natriuretic peptide mRNA and immunoreactivity in the rat heart and human atrial appendage. Proc. Natl. Acad. Sci. USA 84: 6760–6764.

    Article  PubMed  CAS  Google Scholar 

  • Hamid QA, Bishop AE, Springall DR, Adams C, Giaid A, Denny P, Ghatei M, Legon S, Cutitta F, Rode J, Spindel E, Bloom SR and Polak JM (1989) Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. Cancer 63: 266–271.

    Article  PubMed  CAS  Google Scholar 

  • Hofler H, Putz B, Rurhi C, Wirnsberger G, Klimpfinger M and Smolle J (1987) Simultaneous localization of calcitonin mRNA and peptide in a medullary thyroid carcinoma. Virchow. Arch. B 54: 144–151.

    Article  CAS  Google Scholar 

  • Hopman AHN, Wiegant J, Raap AK, L andegent JE, van der Ploeg M and van Dujin P (1986) Bi-colour detection of two target DNAs by non-radioactive in situ hybridization. Histochemistry 85: 1–4.

    Article  PubMed  CAS  Google Scholar 

  • Inoue A, Yanagisawa M, Kimura 5, Kaswya Y, Miyauchi T, Goto T and Masaki T (1989) The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proc. Natl. Acad. Sci. USA 86: 2863–2867.

    Article  PubMed  CAS  Google Scholar 

  • Lawrence JB, Villnave CA and Singer RH (1988) Sensitive, high resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in lymphoma line. Cell 52: 51–61.

    Article  PubMed  CAS  Google Scholar 

  • Leuhersen KR and Baum MP (1987) In vitro synthesis of biotinylated RNA probes from A-T rich templates: problems and solutions. BioTech. 5: 660–670.

    Google Scholar 

  • McCabe JT, Morrell JI and Pfaff DW (1986) In situ hybridization as a quantitative autoradiographic method. In: In Situ Hybridization in Brain. (Uhl GR ed.), pp. 73–96, Plenum Press, New York.

    Chapter  Google Scholar 

  • McCafferty J, Cresswell L, Alldus C, Terenghi G and Fallon R (1989) A shortened protocol for in situ hybridization to mRNA using radiolabelled RNA probes. Techniques 1: 171–182.

    Google Scholar 

  • Melton D, Kneg P, Rebagliati M, Maniatis T, Zinn K and Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmid containing a bacteriophage SP6 promoter. Nuc. Acids Res. 12: 7035–7050.

    Article  CAS  Google Scholar 

  • Mitchell AR, Ambros P, Gosden JR, Morten JEN and Porteous DJ (1986) Gene mapping and physical arrangement of human chromatin in transformed hybrid cells: fluorescent and autoradiographic in situ hybridization compared. Somatic Cell Mol. Genet. 12: 313324.

    Article  Google Scholar 

  • Niedobitek G, Finn T, Herbst H, Bornhoft G, Gerdes J and Stein H (1988) Detection of viral DNA by in situ hybridization using bromodeoxyuridine-labelled DNA probes. Am. J. Pathol. 131: 1–4.

    PubMed  CAS  Google Scholar 

  • Nunez W, Davenport AP, Emson PC and Brown MJ (1989) A quantitative in situ hybridization method using computer-assisted image analysis. Biochem J. 262: 121–127.

    Google Scholar 

  • Polak JM and Van Noorden S (1986) In: Immunocytochemistry — Modern Methods and Applications. 2nd edn. (Polak JM and Van Noorden S eds), Wright, Bristol.

    Google Scholar 

  • Polak JM (1989) Regulatory peptides, Birkhausen Verlag, Basel.

    Book  Google Scholar 

  • Shivers BD, Harl and RE, Pfaff DW and Schachter BS (1986a) Combination of immunocytochemistry and in situ hybridization in the same tissue section of rat pituitary. J. Histochem. Cytochem. 34: 39–43.

    Article  PubMed  CAS  Google Scholar 

  • Shivers BD, Harlan RE, Romano GJ, Howells RD and Pfaff DW (1986b) Cellular localization and regulation of proenkephalin mRNA in rat brain. In: In Situ Hybridization in Brain. (Uhl GR ed.), pp. 3–20, Plenum Press, New York.

    Chapter  Google Scholar 

  • Singer RH, Lawrence JB and Villnave C (1986) Optimisation of in situ hybridization using isotopic and non-isotopic detection methods. BioTechniques 4: 230–250.

    CAS  Google Scholar 

  • Springall DR, Bhatnagar M, Wharton J, Hamid Q, Gulbenkian S, Hedges M, Meleagros L, Bloom SR and Polak JM (1988) Expression of the atrial natriuretic peptide gene in the cardiac muscle of rat extrapulmonary and intrapulmonary veins. Thorax 43: 44–52.

    Article  PubMed  CAS  Google Scholar 

  • Steel JH, Hamid Q, Van Noorden S, Jones P, Denny P, Burrin J, Legon S, Bloom SR and Polak JM (1988a) Combined use of in situ hybridization and immunocytochemistry for the investigation of prolactin gene expression in immature, pubertal, pregnant, lactating and ovarectomized rats. Histochemistry 89: 75–80.

    Article  PubMed  CAS  Google Scholar 

  • Steel JH, Hamid Q, Van Noorden S, Ch andrachud L, Jones P, Denny P, Burrin J, McNicol AM, Legon S, Bloom SR and Polak JM (1988b) Changes in prolactin and pro-opiomelanocortin messenger RNA in rat pituitary as shown by in situ hybridization. In: Neuroendocrine Perspectives, Vol. 6, (Scarlon MF, Wass JAH eds), Springer-Verlag, New York.

    Google Scholar 

  • Steel JH, O’Halloran DJ, Jones PM, Chin WW, Bloom SR and Polak JM (1989) Simultaneous immunocytochemistry and in situ hybridization of beta thyroid-stimulating hormone and its messenger ribonucleic acid in euthyroid and hypothyroid rat pituitary. Mol. Cell. Probes, 4: 385–396s.

    Article  Google Scholar 

  • Terenghi G, Polak JM, Ghatei MA, Mulderry PK, Butler JM, Unger WG and Bloom SR (1985) Distribution and origin of calcitonin generelated peptide immunoreactivity in the sensory innervation of the mammalian eye. J. Comp. Neurol. 233: 506–516.

    Article  PubMed  CAS  Google Scholar 

  • Terenghi G, Polak JM, Hamid Q, O’Brien E, Denny P, Legon S, Dixon J, Minth CD, Palay SL, Yasargil G and Chan-Palay V (1987) Localization of neuropeptide Y mRNA in neurons of human cerebral cortex by means of in situ hybridization with a complementary RNA probe. Proc. Natl. Acad. Sci. USA 84: 7315–7318.

    Article  PubMed  CAS  Google Scholar 

  • Terenghi G and Fallon RA (1990) Techniques and applications of in situ hybridization. In: Current Topics in Pathology — Pathology of the Nucleus, Vol. 82, (Underwood JCE ed.), pp. 289–337, Springer-Verlag, Amsterdam.

    Google Scholar 

  • Van der Ploeg M, L andegent JE, Hopman HHN and Raap AK (1986) Nonautoradiographic hybridocytochemistry. J. Histochem. Cytochem. 34: 126–133.

    Google Scholar 

  • Varndell IM, Polak JM, Sikri KL, Minth CD, Bloom SR and Dixon JE (1984) Visualisation of mRNA directing peptide synthesis by in situ hybridization using a novel single str anded cDNA probe: potential for the investigation of gene expression and endocrine cell activity. Histochemistry 81: 597–601.

    Article  PubMed  CAS  Google Scholar 

  • Wertmur JG, Ruyecham WT and Donthart RJ (1981) Denaturation and renaturation of Penicillin crysogenum mycophage double-str anded ribonucleic acid in tetraalkylammonium salt solution. Biochemistry 20: 2999–3002.

    Article  Google Scholar 

  • Wolf S, Quaas R, Hahu U and Witting B (1987) Synthesis of highly radioactiyely labelled RNA hybridization probes from synthetic single-str anded DNA oligonucleotides. Nuc. Acids Res. 15: 858.

    Article  Google Scholar 

  • Wolfson B, Manning RW, David LG, Arentzen R and Baldino F Jr (1985) Colocalization of corticotropin releasing factor and vasopressin mRNA in neurons after adrenalectomy. Nature 315: 59–63.

    Article  PubMed  CAS  Google Scholar 

  • Young WS and Kuhar MJ (1986) Quantitative in situ hybridization and determination of mRNA content. In: In Situ Hybridization in Brain. (Uhl GR ed.), pp. 243–248, Plenum Press, New York.

    Google Scholar 

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Author information

Authors and Affiliations

  1. Histochemistry Department, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, W12 0NN, London, UK

    Giorgio Terenghi & Julia M. Polak

Authors
  1. Giorgio Terenghi
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  2. Julia M. Polak
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Editors and Affiliations

  1. Department of Biochemistry, Charing Cross and Westminster Medical School, London, UK

    G. R. Coulton  & J. de Belleroche  & 

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© 1992 Springer Science+Business Media Dordrecht

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Terenghi, G., Polak, J.M. (1992). Use of comparative in situ hybridization and immunocytochemistry for the study of regulatory peptides. In: Coulton, G.R., de Belleroche, J. (eds) In Situ Hybridization: Medical Applications. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-2984-8_2

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  • DOI: https://doi.org/10.1007/978-94-011-2984-8_2

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-5326-6

  • Online ISBN: 978-94-011-2984-8

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