Cellular Localization of Induced Human Interferon-beta MRNA by Non-radioactive in Situ Hybridization

Overview

Journal Histochemistry

Specialty Biochemistry

Date 1989 Jan 1

PMID 2732096

Citations 3

Authors

H Multhaupt

G Gross

P Fritz

K Kohler

Affiliations

Soon will be listed here.

Abstract

Induced interferon-beta (IFN-beta) mRNA was localized in human FS-4 fibroblasts by in situ hybridization using biotinylated probes. The hybridization sites were detected by incubation with a nick-translated genomic DNA probe (1.8 kb) via streptavidin-colloidal gold followed by silver contrast enhancement. The positive signals were observed by reflection-contrast light microscopy. IFN-beta mRNA was transiently induced by poly r(I): r(C) in fibroblasts 2-4 h after induction. Induction in the presence of cycloheximide and actinomycin D (superinduction conditions) exhibited an enhanced level of IFN-beta mRNA with a maximum at 4-8 h. The kinetics of the IFN-beta mRNA expression in the cytoplasm as revealed by in situ hybridization proved to be compatible with the results of Northern blotting experiments of total cellular RNA.

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References

Wehner K, Fritz P, Kohler K, Multhaupt H . Visualization and quantitation of poly A+-mRNA by using in situ hybridization with biotinylated oligonucleotides. Acta Histochem Suppl. 1989; 37:83-8. View

Field A, TYTELL A, Lampson G, HILLEMAN M . Inducers of interferon and host resistance. II. Multistranded synthetic polynucleotide complexes. Proc Natl Acad Sci U S A. 1967; 58(3):1004-10. PMC: 335739. DOI: 10.1073/pnas.58.3.1004. View

Landegent J, Jansen in de Wal N, van Ommen G, Baas F, de Vijlder J, VAN DUIJN P . Chromosomal localization of a unique gene by non-autoradiographic in situ hybridization. Nature. 1985; 317(6033):175-7. DOI: 10.1038/317175a0. View

Denhardt D . A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966; 23(5):641-6. DOI: 10.1016/0006-291x(66)90447-5. View

Gall J, Pardue M . Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969; 63(2):378-83. PMC: 223575. DOI: 10.1073/pnas.63.2.378. View

Singer R, Ward D . Actin gene expression visualized in chicken muscle tissue culture by using in situ hybridization with a biotinated nucleotide analog. Proc Natl Acad Sci U S A. 1982; 79(23):7331-5. PMC: 347333. DOI: 10.1073/pnas.79.23.7331. View

Raj N, Pitha P . Analysis of interferon mRNA in human fibroblast cells induced to produce interferon. Proc Natl Acad Sci U S A. 1981; 78(12):7426-30. PMC: 349280. DOI: 10.1073/pnas.78.12.7426. View

HOEFSMIT E, Korn C, Blijleven N, Ploem J . Light microscopical detection of single 5 and 20 nm gold particles used for immunolabelling of plasma membrane antigens with silver enhancement and reflection contrast. J Microsc. 1986; 143(Pt 2):161-9. DOI: 10.1111/j.1365-2818.1986.tb02774.x. View

Hayashi S, Gillam I, Delaney A, TENER G . Acetylation of chromosome squashes of Drosophila melanogaster decreases the background in autoradiographs from hybridization with [125I]-labeled RNA. J Histochem Cytochem. 1978; 26(8):677-9. DOI: 10.1177/26.8.99471. View

10.

Holgate C, Jackson P, Cowen P, Bird C . Immunogold-silver staining: new method of immunostaining with enhanced sensitivity. J Histochem Cytochem. 1983; 31(7):938-44. DOI: 10.1177/31.7.6189883. View

11.

Bauman J, Wiegant J, Borst P, VAN DUIJN P . A new method for fluorescence microscopical localization of specific DNA sequences by in situ hybridization of fluorochromelabelled RNA. Exp Cell Res. 1980; 128(2):485-90. DOI: 10.1016/0014-4827(80)90087-7. View

12.

Landegent J, Jansen in de Wal N, Ploem J, VAN DER PLOEG M . Sensitive detection of hybridocytochemical results by means of reflection-contrast microscopy. J Histochem Cytochem. 1985; 33(12):1241-6. DOI: 10.1177/33.12.2415575. View

13.

Goodbourn S, Maniatis T . Overlapping positive and negative regulatory domains of the human beta-interferon gene. Proc Natl Acad Sci U S A. 1988; 85(5):1447-51. PMC: 279788. DOI: 10.1073/pnas.85.5.1447. View

14.

Berger S, Hitchcock M, Zoon K, Birkenmeier C, Friedman R, Chang E . Characterization of interferon messenger RNA synthesis in Namalva cells. J Biol Chem. 1980; 255(7):2955-61. View

15.

Gross G, Mayr U, BRUNS W, Grosveld F, Dahl H, Collins J . The structure of a thirty-six kilobase region of the human chromosome including the fibroblast interferon gene IFN-beta. Nucleic Acids Res. 1981; 9(11):2495-507. PMC: 326866. DOI: 10.1093/nar/9.11.2495. View

16.

Cavalieri R, Havell E, Vilcek J, Pestka S . Induction and decay of human fibroblast interferon mRNA. Proc Natl Acad Sci U S A. 1977; 74(10):4415-9. PMC: 431953. DOI: 10.1073/pnas.74.10.4415. View

17.

Vilcek J, Ng M . Post-transcriptional control of interferon synthesis. J Virol. 1971; 7(5):588-94. PMC: 356167. DOI: 10.1128/JVI.7.5.588-594.1971. View

18.

Raj N, Pitha P . Two levels of regulation of beta-interferon gene expression in human cells. Proc Natl Acad Sci U S A. 1983; 80(13):3923-7. PMC: 394171. DOI: 10.1073/pnas.80.13.3923. View

19.

Varndell I, Polak J, Sikri K, Minth C, Bloom S, Dixon J . Visualisation of messenger RNA directing peptide synthesis by in situ hybridisation using a novel single-stranded cDNA probe. Potential for the investigation of gene expression and endocrine cell activity. Histochemistry. 1984; 81(6):597-601. DOI: 10.1007/BF00489541. View

20.

Langer P, Waldrop A, Ward D . Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981; 78(11):6633-7. PMC: 349103. DOI: 10.1073/pnas.78.11.6633. View