A synthetic oligonucleotide probe encoding for atrial natriuretic peptide detects specific mRNA transcripts in rat heart but not brain using in situ hybridization histochemistry

1990 ◽  
Vol 93 (5) ◽  
pp. 519-523 ◽  
Author(s):  
E. L. Sutin ◽  
P. Montpied ◽  
D. M. Jacobowitz
1994 ◽  
Vol 42 (9) ◽  
pp. 1271-1276 ◽  
Author(s):  
M Numata ◽  
T Ono ◽  
S Iseki

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.


Author(s):  
MR Islam

The aim of the present study to explore the convenient method of in situ hybridization histochemistry for neuronal tissues studies. The glutamate receptor 1 (GluR1) 35S labeled oligonucleotide probe was used for in situ hybridization histochemistry. The glutamate receptor 1 mRNA expression was found abundantly in the dentate gyrus, Ammon`s horn, and subiculum of the rat hippocampus. In comparison to immunohistochemistry as well as use several types of probes like double strand DNA probe, single strand DNA probe, and RNA probe, the most convenient is oligonucleotide probe for the rapid assessment of the expression of a cloned ion channel by in situ hybridization histochemistry. Key words: Oligonucleotide probe, autoradiogram, hippocampus, rat DOI = 10.3329/bjvm.v5i1.1327 Bangl. J. Vet. Med. (2007). 5 (1 & 2): 111-114


1994 ◽  
Vol 42 (9) ◽  
pp. 1277-1283 ◽  
Author(s):  
E Mezey ◽  
B J Hoffman ◽  
G Harta ◽  
M Palkovits ◽  
J Northup

In this study we examined the cause of unusually intense signals obtained in immune cells by in situ hybridization histochemistry using 35S-labeled oligonucleotides. We verified that the phenomenon is an amplification of a specific signal due to a series of chemical interactions after the probe binds to a specific mRNA in the tissue. The presence of oxidative enzymes in the tissue seems to be necessary for this reaction to occur. Therefore, most cells of the immune system (e.g., macrophages, neutrophil and eosinophil leukocytes), being rich in oxidative enzymes, will show some signal amplification. The intensification of the signal can be avoided if MgCl2 is substituted for CoCl2 in the synthesis of [35S]-thiophosphate-labeled probes, if 2,3-dimercaptopropanol [British anti-Lewisite (BAL)] is added to the hybridization buffer, or if [33P]-phosphate is used instead of [35S]-thiophosphate in the labeling of the probes.


1988 ◽  
Vol 66 (8) ◽  
pp. 1113-1121 ◽  
Author(s):  
V. K. M. Han ◽  
A. J. D'Ercole ◽  
D. C. Lee

Transforming growth factors (TGFs) are polypeptides that are produced by transformed and tumour cells, and that can confer phenotypic properties associated with transformation on normal cells in culture. One of these growth-regulating molecules, transforming growth factor alpha (TGF-α), is a 50 amino acid polypeptide that is related to epidermal growth factor (EGF) and binds to the EGF receptor. Previous studies have shown that TGF-α is expressed during rodent embryogenesis between 7 and 14 days gestation. To investigate the cellular sites of TGF-α mRNA expression during development, we have performed Northern analyses and in situ hybridization histochemistry on the conceptus and maternal tissues at various gestational ages. Contrary to previous reports, both Northern analyses and in situ hybridization histochemistry indicate that TGF-α mRNA is predominantly expressed in the maternal decidua and not in the embryo. Decidual expression is induced following implantation, peaks at day 8, and declines through day 15 when the decidua is being resorbed. In situ hybridization revealed that expression of TGF-α mRNA is highest in the region of decidua adjacent to the embryo and is low or nondetectable in the uterus, placenta, and embryo. In addition, we could not detect TGF-α mRNA expression in other maternal tissues, indicating that the induction of TGF-α transcripts in the decidua is tissue specific, and not a pleiotropic response to changes in hormonal milieu that occur during pregnancy. The developmentally regulated expression of TGF-α mRNA in the decidua, together with the presence of EGF receptors in this tissue, suggests that this peptide may stimulate mitosis and angiogenesis locally by an autocrine mechanism. Because EGF receptors are also present in the embryo and placenta, TGF-α may act on these tissues by a paracrine or endocrine mechanism.


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