Quantitative detection of alternatively spliced transcripts of the aromatase cytochrome P450 (CYP19) gene in aromatase-expressing human cells by competitive RT-PCR. 1995

V R Agarwal, and S E Bulun, and E R Simpson
Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, USA.

C19 steroids are converted to oestrogens in a number of tissues by a specific form of cytochrome P450, namely aromatase P450 (P450arom; the product of the CYP19 gene). The human CYP19 gene comprises nine coding exons, II-X. The tissue-specific expression is determined by the use of tissue-specific promoters, which give rise to P450arom transcripts with unique 5'-untranslated sequences. The majority of the transcripts present in the ovary contain promoter-II-specific sequences, while transcripts in the placenta contain exon I.1. Transcripts in adipose tissue possess exon I.3 and exon I.4. Also, the distribution of alternative transcripts in adipose stromal cells depends on the culture conditions. Therefore, a competitive RT-PCR method was designed to quantitatively detect alternatively spliced transcripts present in various tissues and cells maintained in different culture conditions. Specific synthetic transcripts with different 5' termini (exon I.3, exon I.4 and promoter-II-specific sequences) and the coding region were used as internal standards. This competitive RT-PCR method was used to quantitatively detect three 5' termini, i.e. promoter-II-specific sequence, exon I.3 and exon I.4, in transcripts in human adipose stromal cells and ovarian granulosa cells in primary culture. The quantity of total P450arom transcripts was judged by amplifying the coding region. We were also able to quantify rare transcripts which could not be detected previously by Northern analysis.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009928 Organ Specificity Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen. Tissue Specificity,Organ Specificities,Specificities, Organ,Specificities, Tissue,Specificity, Organ,Specificity, Tissue,Tissue Specificities
D012015 Reference Standards A basis of value established for the measure of quantity, weight, extent or quality, e.g. weight standards, standard solutions, methods, techniques, and procedures used in diagnosis and therapy. Standard Preparations,Standards, Reference,Preparations, Standard,Standardization,Standards,Preparation, Standard,Reference Standard,Standard Preparation,Standard, Reference
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003907 Dexamethasone An anti-inflammatory 9-fluoro-glucocorticoid. Hexadecadrol,Decaject,Decaject-L.A.,Decameth,Decaspray,Dexasone,Dexpak,Hexadrol,Maxidex,Methylfluorprednisolone,Millicorten,Oradexon,Decaject L.A.
D004790 Enzyme Induction An increase in the rate of synthesis of an enzyme due to the presence of an inducer which acts to derepress the gene responsible for enzyme synthesis. Induction, Enzyme
D005091 Exons The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA. Mini-Exon,Exon,Mini Exon,Mini-Exons
D005260 Female Females
D005576 Colforsin Potent activator of the adenylate cyclase system and the biosynthesis of cyclic AMP. From the plant COLEUS FORSKOHLII. Has antihypertensive, positive inotropic, platelet aggregation inhibitory, and smooth muscle relaxant activities; also lowers intraocular pressure and promotes release of hormones from the pituitary gland. Coleonol,Forskolin,N,N-Dimethyl-beta-alanine-5-(acetyloxy)-3-ethenyldodecahydro-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-1H-naphtho(2,1-b)pyran-6-yl Ester HCl,NKH 477,NKH-477,NKH477
D006107 Granulosa Cells Supporting cells for the developing female gamete in the OVARY. They are derived from the coelomic epithelial cells of the gonadal ridge. Granulosa cells form a single layer around the OOCYTE in the primordial ovarian follicle and advance to form a multilayered cumulus oophorus surrounding the OVUM in the Graafian follicle. The major functions of granulosa cells include the production of steroids and LH receptors (RECEPTORS, LH). Cell, Granulosa,Cells, Granulosa,Granulosa Cell

Related Publications

V R Agarwal, and S E Bulun, and E R Simpson
March 2003, Brain research. Brain research protocols,
V R Agarwal, and S E Bulun, and E R Simpson
October 2001, European journal of clinical investigation,
V R Agarwal, and S E Bulun, and E R Simpson
June 1996, The Journal of clinical endocrinology and metabolism,
V R Agarwal, and S E Bulun, and E R Simpson
December 1994, The Journal of clinical endocrinology and metabolism,
V R Agarwal, and S E Bulun, and E R Simpson
January 2008, Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association,
V R Agarwal, and S E Bulun, and E R Simpson
February 2000, Carcinogenesis,
V R Agarwal, and S E Bulun, and E R Simpson
February 2004, Seminars in reproductive medicine,
V R Agarwal, and S E Bulun, and E R Simpson
February 1994, Molecular endocrinology (Baltimore, Md.),
V R Agarwal, and S E Bulun, and E R Simpson
February 1994, Journal of immunology (Baltimore, Md. : 1950),
V R Agarwal, and S E Bulun, and E R Simpson
January 1996, The Journal of steroid biochemistry and molecular biology,
Copied contents to your clipboard!