scholarly journals Intracellular Targeting of Calmodulin mRNAs in Primary Hippocampal Cells

2003 ◽  
Vol 51 (4) ◽  
pp. 541-544 ◽  
Author(s):  
Elod Kortvely ◽  
Szilvia Varszegi ◽  
Arpad Palfi ◽  
Karoly Gulya
Keyword(s):  
In Situ Hybridization ◽  
Glial Cells ◽  
Cultured Cells ◽  
Intracellular Distribution ◽  
Low Level ◽  
Rna Granules ◽  
Intracellular Targeting ◽  
Dendritic Mrna ◽  
Mrna Targeting

We investigated the intracellular distribution of the mRNAs corresponding to the three non-allelic CaM genes in cultured hippocampal cells by in situ hybridization with digoxigenin-labeled gene-specific riboprobes. In neurons the perikaryon was heavily stained and strong dendritic mRNA targeting was detected for all three CaM genes. The color labeling exhibited a punctate distribution, suggesting that CaM mRNAs are transported in RNA granules. Immunocytochemistry for S100 demonstrated that glial cells express CaM mRNAs at a very low level. A minority of the cultured cells were negative for either labeling.

scholarly journals Detection of v- and c-erbB oncogene mRNA in cultured cells by in situ hybridization.

1990 ◽  
Vol 52 (1) ◽  
pp. 175-178
Author(s):  
Toshio IKEDA ◽  
Yasuhiro YOSHIKAWA ◽  
Kazuya YAMANOUCHI
Keyword(s):  
In Situ Hybridization ◽  
Cultured Cells

scholarly journals Intracellular distribution of histone mRNAs in human fibroblasts studied by in situ hybridization.

1988 ◽  
Vol 85 (2) ◽  
pp. 463-467 ◽  
Author(s):  
J. B. Lawrence ◽  
R. H. Singer ◽  
C. A. Villnave ◽  
J. L. Stein ◽  
G. S. Stein
Keyword(s):  
In Situ Hybridization ◽  
Human Fibroblasts ◽  
Intracellular Distribution ◽  
Histone Mrnas

Methodologies for specific intron and exon RNA localization in cultured cells by haptenized and fluorochromized probes

1993 ◽  
Vol 104 (4) ◽  
pp. 1187-1197 ◽  
Author(s):  
R.W. Dirks ◽  
F.M. van de Rijke ◽  
S. Fujishita ◽  
M. van der Ploeg ◽  
A.K. Raap
Keyword(s):  
In Situ Hybridization ◽  
Cell Lines ◽  
Cultured Cells ◽  
Direct Detection ◽  
Housekeeping Genes ◽  
Housekeeping Gene ◽  
Immediate Early ◽  
High Signal ◽  
Pretreatment Conditions

We have determined optimal conditions for the detection of mRNA sequences in cultured cells by nonradioactive in situ hybridization. For this purpose a number of different cell lines have been used: rat 9G cells for the detection of human cytomegalovirus immediate early mRNA, and HeLa as well as 5637 carcinoma cells for the detection of housekeeping gene mRNAs. Extensive optimization of fixation and pretreatment conditions revealed that most intense hybridization signals are obtained when cells are grown on glass microscope slides, fixed with a mixture of formaldehyde and acetic acid, pretreated with pepsin and denatured prior to hybridization. In addition, we also studied the potential of fluorochromized probes for the direct detection of multiple RNA sequences. The optimized in situ hybridization procedure revealed that immediate early mRNA transcripts are, in addition to a cytoplasmic localization, localized within nuclei of rat 9G cells. Double hybridization experiments showed that intron and exon sequences colocalize within the main nuclear signal. In addition, the presence of small, intron-specific, fluorescent spots scattered around the main nuclear signals indicates that intron sequences which are spliced out can be visualized. Additional information about the functioning of cells could be obtained by the detection of mRNA simultaneously with bromodeoxyuridine, incorporated during S-phase, or its cognate protein. The sensitivity of these methods is such that mRNAs of abundantly expressed housekeeping genes can be detected in a variety of cell lines with high signal to noise ratios.

scholarly journals Significance of HER2 Low-Level Copy Gain in Barrett's Cancer: Implications for Fluorescence In situ Hybridization Testing in Tissues

2007 ◽  
Vol 13 (17) ◽  
pp. 5115-5123 ◽  
Author(s):  
Sandra Rauser ◽  
Roland Weis ◽  
Herbert Braselmann ◽  
Marcus Feith ◽  
Hubert J. Stein ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Low Level ◽  
Barrett’S Cancer

scholarly journals Gene expression analysis by non-radioactive RNA-RNA in situ hybridization techniques

2004 ◽  
Vol 23 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Danijela Drakulic ◽  
Milena Stevanovic ◽  
Gordana Nikcevic
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Cultured Cells ◽  
Specific Gene ◽  
Rna In Situ Hybridization ◽  
Sox Gene ◽  
Labeled Rna ◽  
Set Up

RNA-RNA in situ hybridization is a reliable method for studying tissue and cell specific gene expression, which enables visualization of labeled antisense RNA probe hybridized to specific mRNA. In this study we employed non-radioactive RNA-RNA in situ hybridization using biotin- or digoxigenin-labeled RNA probes in order to detect SOX gene expression in carcinoma cell lines. By this approach we confirmed results obtained by Northern blot analysis, where the presence of SOX2 mRNA in NT2/D1 and SOX14 mRNA in HepG2 cells has been established. Our aim was to set up RNA-RNA in situ hybridization method in in vitro cultured cells in order to perform further analyses of SOX gene expression on various normal and cancer tissues.

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