A Rapid and Highly Sensitive In Situ mRNA Hybridization Method With Digoxigenin Labeled cRNA Probes

2006 ◽  
Vol 29 (2) ◽  
pp. 91-95
Author(s):  
Yuichi Sato ◽  
Benio Tsuchiya ◽  
Ryo Nagashio ◽  
Shi-Xu Jiang ◽  
Isao Okayasu
Keyword(s):  
Hybridization Method ◽  
Highly Sensitive ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization

scholarly journals Telokin expression is restricted to smooth muscle tissues during mouse development

2001 ◽  
Vol 280 (1) ◽  
pp. C12-C21 ◽  
Author(s):  
B. Paul Herring ◽  
Gary E. Lyons ◽  
April M. Hoggatt ◽  
Patricia J. Gallagher
Keyword(s):  
Smooth Muscle ◽  
Postnatal Development ◽  
Mouse Development ◽  
Muscle Tissues ◽  
Cell Layers ◽  
Mrna Analysis ◽  
Muscle Myosin ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization

Telokin is a 17-kDa protein with an amino acid sequence that is identical to the COOH terminus of the 130-kDa myosin light chain kinase (MLCK). Telokin mRNA is transcribed from a second promoter, located within an intron, in the 3′ region of the MLCK gene. In the current study, we show by in situ mRNA hybridization that telokin mRNA is restricted to the smooth muscle cell layers within adult smooth muscle tissues. In situ mRNA analysis of mouse embryos also revealed that telokin expression is restricted to smooth muscle tissues during embryonic development. Telokin mRNA expression was first detected in mouse gut at embryonic day 11.5; no telokin expression was detected in embryonic cardiac or skeletal muscle. Expression of telokin was also found to be regulated during postnatal development of the male and female reproductive tracts. In both uterus and vas deferens, telokin protein expression greatly increased between days 7 and 14 of postnatal development. The increase in telokin expression correlated with an increase in the expression of several other smooth muscle-restricted proteins, including smooth muscle myosin and α-actin.

scholarly journals Cardiac endothelial cells maintain open chromatin and expression of cardiomyocyte myofibrillar genes

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Nora Yucel ◽  
Jessie Axsom ◽  
Yifan Yang ◽  
Li Li ◽  
Joshua H Rhoades ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Open Chromatin ◽  
Gene Promoters ◽  
Isolated Nuclei ◽  
Parenchymal Cells ◽  
Cardiac Transcription Factors ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization

Endothelial cells (ECs) are widely heterogenous depending on tissue and vascular localization. Jambusaria et al. recently demonstrated that ECs in various tissues surprisingly possess mRNA signatures of their underlying parenchyma. The mechanism underlying this observation remains unexplained, and could include mRNA contamination during cell isolation, in vivo mRNA paracrine transfer from parenchymal cells to ECs, or cell-autonomous expression of these mRNAs in ECs. Here, we use a combination of bulk RNASeq, single-cell RNASeq datasets, in situ mRNA hybridization, and most importantly ATAC-Seq of FACS-isolated nuclei, to show that cardiac ECs actively express cardiomyocyte myofibril (CMF) genes and have open chromatin at CMF gene promoters. These open chromatin sites are enriched for sites targeted by cardiac transcription factors, and closed upon expansion of ECs in culture. Together, these data demonstrate unambiguously that the expression of CMF genes in ECs is cell-autonomous, and not simply a result of technical contamination or paracrine transfers of mRNAs, and indicate that local cues in the heart in vivo unexpectedly maintain fully open chromatin in ECs at genes previously thought limited to cardiomyocytes.

scholarly journals Dealing with the problem of non-specific in situ mRNA hybridization signals associated with plant tissues undergoing programmed cell death

Plant Methods ◽  
2010 ◽  
Vol 6 (1) ◽  
pp. 7 ◽  
Author(s):  
Jaana Vuosku ◽  
Suvi Sutela ◽  
Mira Sääskilahti ◽  
Johanna Kestilä ◽  
Anne Jokela ◽  
...  
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Plant Tissues ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization

Endothelin-1 and its binding sites are upregulated in pressure overload cardiac hypertrophy

1995 ◽  
Vol 268 (5) ◽  
pp. H2084-H2091 ◽  
Author(s):  
M. Arai ◽  
A. Yoguchi ◽  
T. Iso ◽  
T. Takahashi ◽  
S. Imai ◽  
...  
Keyword(s):  
Cardiac Hypertrophy ◽  
Binding Sites ◽  
Pressure Overload ◽  
Sham Operation ◽  
Endothelin 1 ◽  
Important Relationship ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization

The purpose of this study was to determine whether endothelin and endothelin receptors play an important role in the development of cardiac hypertrophy due to pressure overload in vivo. Cardiac hypertrophy was produced by placing a constricting clip around the suprarenal abdominal aorta of rats. Hemodynamic parameters and plasma and ventricular concentrations of endothelin-1 (ET-1) were measured in control unoperated rats, and 30 min, 2 and 6 h, and 1 and 8 days after operation in pressure overload rats and sham-operated rats. The density and dissociation constant of ET-1 binding sites were also measured in control rats and 1 and 8 days after pressure overload and sham operation. Additionally, in situ mRNA hybridization for preproendothelin-1 (preproET-1) mRNA was performed to determine which cells were responsible for increased ET-1 levels. Ventricular ET-1 levels increased markedly on day 8 of pressure overload, whereas plasma ET-1 levels increased transiently only 30 min after operation, quickly returning to control level. In addition, ventricular ET-1 levels on day 8 showed a significant positive correlation with the degree of cardiac hypertrophy. In situ mRNA hybridization revealed that cardiac myocytes expressed preproET-1 mRNA in hypertrophied hearts in vivo. In accord with the elevation of ventricular ET-1 levels, the density of ET-1 binding sites was increased significantly, without affecting their binding affinity, on day 8 of pressure overload. These data are compatible with the hypothesis that increases in locally produced ET-1 and the density of ET-1 binding sites have an important relationship with the development of cardiac hypertrophy in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

In situ mRNA hybridization study of the distribution of choline acetyltransferase in the human brain

1998 ◽  
Vol 806 (1) ◽  
pp. 8-15 ◽  
Author(s):  
Satomi Kasashima ◽  
Youko Muroishi ◽  
Hiroko Futakuchi ◽  
Isao Nakanishi ◽  
Yoshio Oda
Keyword(s):  
Human Brain ◽  
Choline Acetyltransferase ◽  
Hybridization Study ◽  
Mrna Hybridization ◽  
In Situ Mrna Hybridization
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