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  • 1
    Electronic Resource
    Electronic Resource
    Expression of m2 muscarinic acetylcholine receptor mRNA in primary culture of human prostate stromal cells (2000)
    Springer
    Urological research 28 (2000), S. 196-200 
    Details
    ISSN: 1434-0879
    Keywords: Key words Muscarinic acethylcholine receptor subtypes mRNA expression ; Primary culture of human prostate stromal cells ; Reverse transcription polymerase chain reaction ; RNA blotting In situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aim of this study was to investigate the expression of the muscarinic acetylcholine receptor (mAchR) subtypes mRNA in primary culture of human prostate stromal cells using the reverse transcription polymerase chain reaction (RT-PCR), RNA blotting and in situ hybridization (ISH). Using an explant method, we obtained a primary culture of prostate stromal cells from three patients with benign prostatic hypertrophy. Total RNA was extracted using the acid guanidinium method for cDNA synthesis. First-strand cDNA was then used for PCR with primers designed to amplify the fragments of each mAchR subtypes (m1–m5) cDNA sequence. The m2, m3 and m4 subtype expected bands were detected; in particular m2 transcripts was strongly detected in the stromal cell culture. Each of the PCR products were subcloned into the pGEM-T plasmid vector, sequenced and random primer labeled using 32P. Digoxigenin-labeled cRNA probes were synthesized by in vitro transcription. RNA blotting using a m2 muscarinic receptor cDNA probe revealed a 4.5 kb single transcript. However, m3 and m4 probes did not hybridize. Using in situ hybridization (ISH), m2 receptor mRNA signals were detected in several smooth muscle cells. The staining was predominantly localized to the perinuclear cytoplasm. The m3 and m4 probes did not hybridize. These results suggested that m2 receptor subtype plays a role in smooth muscle activity of the human prostate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002400000113
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  • 2
    Electronic Resource
    Electronic Resource
    cDNA cloning of a human RAB26-related gene encoding a Ras-like GTP-binding protein on chromosome 16p13.3 region (2000)
    Springer
    Journal of human genetics 45 (2000), S. 309-314 
    Details
    ISSN: 1435-232X
    Keywords: Key words Ras superfamily of small GTP-binding proteins ; RAB26-related ; Rab26 ; RT-PCR ; RH mapping ; Chromosome 16p13.3 ; Virtual transcribed sequence (VTS)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Members of the RAB protein family are important regulators of vesicular fusion and trafficking. A putative new member of the RAB family of genes was identified through a public database search, and its full-length cDNA was isolated from a human fetal brain cDNA library. The predicted protein product of the gene consists of 190 amino acid residues and has 87% identity with rat Rab26. Thus, we designated this gene as the human RAB26-related gene. Reverse transcription-coupled polymerase chain reaction (RT-PCR) demonstrated that the RAB26-related messenger RNA was predominantly expressed in adult and fetal brain. Furthermore, an RT-PCR experiment for brain subregions showed that the mRNA was highly expressed in the amygdala, cerebellum, caudate nucleus, and hippocampus. By PCR-based analysis with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid panel, the gene was mapped to the chromosome 16p13.3 region between markers WI-7742 and WI-3061. The RAB26-related gene consists of eight exons that span about 44 kb of the genome DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s100380070023
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    Paper (German National Licenses)
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