SIMULTANEOUS DETECTION OF 2 MESSENGER-RNAS IN THE CENTRAL-NERVOUS-SYSTEM - A SIMPLE 2-STEP INSITU HYBRIDIZATION PROCEDURE USING A COMBINATION OF RADIOACTIVE AND NONRADIOACTIVE PROBES

被引：26

作者：

NORMAND, E ^{[1
]}

BLOCH, B ^{[1
]}

机构：

[1] UNIV BORDEAUX 2,HISTOL EMBRYOL LAB,CNRS,URA 1200,146 RUE LEO SAIGNAT,F-33076 BORDEAUX,FRANCE

来源：

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY | 1991年 / 39卷 / 11期

关键词：

INSITU HYBRIDIZATION; MESSENGER RNA; COLOCALIZATION; NEUROPEPTIDE; RECEPTOR; CENTRAL NERVOUS SYSTEM;

D O I：

10.1177/39.11.1918932

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We present here a method enabling the simultaneous detection of two messenger RNAs in tissue sections by use of a two-step in situ hybridization procedure. Tissue sections were hybridized with a radioactive probe and coated with emulsion. The emulsion was processed for development, fixed, and a second hybridization was performed through the emulsion with a biotinylated probe subsequently revealed with streptavidin-alkaline phosphatase. This procedure allows the detection of two mRNAs without loss of signal, removal of the emulsion, or spurious reaction. The simultaneous detection of oxytocin and vasopressin mRNAs in the hypothalamus, and of dopamine receptor and neuropeptide mRNAs in the striatum, demonstrated the efficiency of the procedure. Such a two-step procedure provides a simple and flexible way to make possible comparative analysis of the localization of two mRNAs within the same tissue section.

引用

页码：1575 / 1578

页数：4

共 22 条

[1] INSITU HYBRIDIZATION HISTOCHEMISTRY FOR THE ANALYSIS OF GENE-EXPRESSION IN THE ENDOCRINE AND CENTRAL-NERVOUS-SYSTEM TISSUES - A 3-YEAR EXPERIENCE [J].

BLOCH, B ;

POPOVICI, T ;

LEGUELLEC, D ;

NORMAND, E ;

CHOUHAM, S ;

GUITTENY, AF ;

BOHLEN, P .

JOURNAL OF NEUROSCIENCE RESEARCH, 1986, 16 (01) :183-200

[2] TOPOGRAPHY AND ONTOGENY OF THE NEURONS EXPRESSING VASOPRESSIN, OXYTOCIN, AND SOMATOSTATIN GENES IN THE RAT-BRAIN - AN ANALYSIS USING RADIOACTIVE AND BIOTINYLATED OLIGONUCLEOTIDES [J].

BLOCH, B ;

GUITTENY, AF ;

CHOUHAM, S ;

MOUGIN, C ;

ROGET, A ;

TEOULE, R .

CELLULAR AND MOLECULAR NEUROBIOLOGY, 1990, 10 (01) :99-112

[3]

CHESSELET M, 1990, IN SITU HYBRIDIZATIO

[4] DETECTION OF MESSENGER-RNA MOLECULES CODING FOR NEUROPEPTIDE HORMONES OF THE POND SNAIL LYMNAEA-STAGNALIS BY RADIOACTIVE AND NON-RADIOACTIVE INSITU HYBRIDIZATION - A MODEL STUDY FOR MESSENGER-RNA DETECTION [J].

DIRKS, RW ;

RAAP, AK ;

VANMINNEN, J ;

VREUGDENHIL, E ;

SMIT, AB ;

VANDERPLOEG, M .

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1989, 37 (01) :7-14

[5] DOUBLE-LABELING INSITU HYBRIDIZATION ANALYSIS OF MESSENGERS RNAS FOR CARBONIC ANHYDRASE-II AND MYELIN BASIC-PROTEIN - EXPRESSION IN DEVELOPING CULTURED GLIAL-CELLS [J].

GHANDOUR, MS ;

SKOFF, RP .

GLIA, 1991, 4 (01) :1-10

[6] HISTOLOGICAL DETECTION OF MESSENGER-RNAS WITH BIOTINYLATED SYNTHETIC OLIGONUCLEOTIDE PROBES [J].

GUITTENY, AF ;

FOUQUE, B ;

MOUGIN, C ;

TEOULE, R ;

BLOCH, B .

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1988, 36 (06) :563-571

[7] DETECTION OF 2 VIRAL GENOMES IN SINGLE CELLS BY DOUBLE-LABEL HYBRIDIZATION INSITU AND COLOR MICRORADIOAUTOGRAPHY [J].

HAASE, AT ;

WALKER, D ;

STOWRING, L ;

VENTURA, P ;

GEBALLE, A ;

BLUM, H ;

BRAHIC, M ;

GOLDBERG, R ;

OBRIEN, K .

SCIENCE, 1985, 227 (4683) :189-192

[8] BI-COLOR DETECTION OF 2-TARGET DNAS BY NONRADIOACTIVE INSITU HYBRIDIZATION [J].

HOPMAN, AHN ;

WIEGANT, J ;

RAAP, AK ;

LANDEGENT, JE ;

VANDERPLOEG, M ;

VANDUIJN, P .

HISTOCHEMISTRY, 1986, 85 (01) :1-4

[9]

ICHIMIYA Y, 1989, J NEUROENDOCRINOL, V1, P74

[10] INTRACELLULAR-LOCALIZATION OF MESSENGER-RNAS FOR CYTOSKELETAL PROTEINS [J].

LAWRENCE, JB ;

SINGER, RH .

CELL, 1986, 45 (03) :407-415

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