Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Estrogens are key regulators of sexual differentiation and development in vertebrates. The P450 aromatase (P450arom) is the steroidogenic enzyme responsible for the synthesis of estrogens from androgens. In the adult rat testis, aromatase transcripts and activity have been observed in somatic cells and germ cells, including pachytene spermatocytes (PS) and round spermatids (RS), but little is known concerning regulation of the aromatase gene expression, especially in germ cells. The quality of germ cell preparations was assessed by the absence of androgen-binding protein and stem cell factor transcripts, two specific markers for Sertoli cells. By employing a competitive quantitative reverse transcriptase-polymerase chain reaction technique, we confirmed that germ cells contained P450arom transcripts and demonstrated that the aromatase gene was up-regulated by cAMP. Conversely, transforming growth factor (TGF) beta1 inhibited Cyp19 gene expression in a dose- and a time-dependent manner in both PS and RS. The addition of tumor necrosis factor (TNF) alpha to purified germ cells induced an increase of the amount of P450arom mRNA in PS, although an inhibitory effect was observed in RS. When PS were treated with dexamethasone (Dex), a similar enhancement of the aromatase transcript level was observed, whereas an inhibitory effect was recorded for RS. Furthermore, in either TGFbeta1- or TNFalpha-treated germ cells, the addition of Dex stimulated the aromatase gene transcription. Experiments using 5' rapid amplification of cDNA ends suggested that promoter PII is mainly concerned in the regulation of the aromatase gene expression in germ cells of adult male rats; however, the presence of other promoters could not be excluded.
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PMID:Regulation of aromatase gene expression in purified germ cells of adult male rats: effects of transforming growth factor beta, tumor necrosis factor alpha, and cyclic adenosine 3',5'-monosphosphate. 1270 Jan 95

The expression of aromatase, the enzyme that transforms testosterone into estradiol, was analyzed by reverse transcriptase polymerase chain reaction in the inferior olive of adult male rats. The expression of this messenger in the inferior olive suggests that this brain area may be able to synthesize estradiol. The neuroprotective role of estradiol in the inferior olive was then assessed in a model of cerebellar ataxia, achieved by the ip administration of 3-acetylpyridine (3-AP). In a first experiment, male Wistar rats were orchidectomized to diminish the plasmatic levels of testosterone, the direct precursor of estradiol. Immediately after castration, animals were implanted with a silicone tube that was either empty or filled with estradiol. One week later, animals were injected with 3-AP. Estradiol treatment resulted in a significant reduction in neuronal death in the olive. In a second experiment, animals were treated with fadrozole, an aromatase inhibitor, to assess the role of endogenous estradiol formation in neuroprotection. The results show that the inhibition of aromatase activity, and therefore the decrease in endogenous estrogen formation, increases the death in inferior olive. In conclusion, this study indicates that the inferior olive is a steroidogenic tissue and that olivary neurons are protected by exogenous and endogenous estradiol.
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PMID:Endogenous estrogen formation is neuroprotective in model of cerebellar ataxia. 1277 2

Recent data indicate that estrogens locally produced in the brain by aromatization of androgens could be important for neurogenesis and brain repair. In this respect, fish are interesting because of the extremely high aromatase activity of their brain. In this study, the rainbow trout brain aromatase was cloned and riboprobes were used to map the distribution of cells expressing the corresponding mRNAs. A very strong hybridization signal was detected in the pituitary and in cells bordering the ventricles in the telencephalon and ventral diencephalon, with the highest expression in the preoptic area and hypothalamus. A weaker signal was detected in the ependymal layer bordering the torus semicircularis and optic tectum. This localization was fully confirmed by immunohistochemistry using antibodies against a teleost aromatase. In addition, this antibody showed that aromatase expression in fact corresponds to radial glial cells because immunoreactive cells had long cytoplasmic processes extending toward the pial surface. Because brain aromatase was shown to be upregulated by estradiol in fish, the distribution of aromatase mRNAs was compared with that of rainbow trout estrogen receptor alpha (rtERalpha) on adjacent sections. Although the highest aromatase expression was found in regions expressing rtERalpha, no obvious coexpression was found, as rtERalpha was never observed in radial cells. However, reverse transcriptase-polymerase chain reaction experiments performed on brain cell cultures enriched in glial cells suggest that a weak expression of rtERalpha in glial cells cannot be excluded. The possible role of the high brain aromatase content in fish could be related to the continuous growth of their central nervous system during adulthood.
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PMID:Distribution of aromatase mRNA and protein in the brain and pituitary of female rainbow trout: Comparison with estrogen receptor alpha. 1279 42

A number of conditions related to sex-reversal in boys and men and precocious puberty in girls are caused by estrogen-secreting adrenal tumors. In these tumors, cytochrome P450 aromatase (aromatase) that is encoded in the CYP19 gene is expressed at high levels. To investigate the molecular mechanism of aromatase expression in these adrenal tumors, we characterized the activity, gene transcript and genomic promoter region of aromatase in the human adrenocortical carcinoma cell line H295R. Aromatase activity and the transcript of the CYP19 gene were highly up-regulated by forskolin, but not by dexamethasone. The results from exon I-specific reverse transcriptase (RT)-PCR and the transfection of reporter constructs suggested that promoter I.3 and promoter II were activated in H295R. Deletion and mutation analysis suggested that cAMP response element-like sequence (CLS) and steroidogenic factor-1 (SF-1) motif, were critical for the activation of promoter II. The results of this work should provide the basis for the molecular analysis of aromatase expression in adrenocortical cells.
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PMID:Forskolin up-regulates aromatase (CYP19) activity and gene transcripts in the human adrenocortical carcinoma cell line H295R. 1470 51

Mutations in the forkhead transcription factor gene FOXL2 are involved in ovarian failure, which occurs in human BPES syndrome. This syndrome presents a sexually dimorphic expression, specific to the ovary in several vertebrates. We cloned the open reading frame of chicken FOXL2 (cFoxL2) and studied cFoxL2 expression in developing gonads and during adulthood to examine the role of FOXL2 in ovarian differentiation and function in birds. The spatial and temporal dynamics of cFoxL2 and aromatase expression were analyzed in parallel by using real-time quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry in attempt to investigate the possible role of cFoxL2 in the regulation of aromatase. The expression patterns of cFoxL2 and aromatase transcripts were highly correlated during the sex-differentiation period (4.7-12.7 days of incubation). Aromatase and cFoxL2 proteins were colocalized in the medullar part of female gonads on embryonic day 14. Fourteen days after hatching, cFoxL2 protein was mainly detected in granulosa cells of developing follicles. In adult ovary follicular envelopes, apart from granulosa cells, cFoxL2 transcript and protein were detected at lower levels in theca cells where aromatase was present. A high level of cFoxL2 transcription was also observed in maturing and ovulated oocytes. Our results confirm that FoxL2 is an early regulator of ovarian development in birds and may be involved in aromatase transcription regulation.
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PMID:Isolation of chicken homolog of the FOXL2 gene and comparison of its expression patterns with those of aromatase during ovarian development. 1551 86

We have previously reported that the repeated electroacupuncture (EA) stimulation significantly increased the concentrations of circulating estradiol and restored the depressed function of the hypothalamus-pituitary-ovary axis (HPOA) in ovariectomized (OVX) rats. We hypothesize that extragonadal aromatization in specific brain areas might be responsible for these changes. Thus, various assays, including radiometric assay, Western blot, and reverse transcriptase polymerase chain reaction (RT-PCR), were employed to determine the aromatization in the hypothalamus of rats that received both OVX and electroacupuncture (OVX + EA). The results showed that EA significantly increased the aromatase activity as well as the expressions of its mRNA and protein (P < 0.05) in the OVX rats. These results suggest that EA enhances brain aromatization, which might contribute to influence the function of gonadotropin-releasing hormone (GnRH) neurons and promote the hypofunction of the HPOA in the ovariectomized rats.
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PMID:Electroacupuncture stimulates hypothalamic aromatization. 1577 65

When larvae of the salamander Hynobius retardatus were reared at a high temperature (28 degrees C) during their thermosensitive period (TSP=15-30 days after hatching), all larvae developed to phenotypic females irrespective of their genetic sexes. Hynobius P450 aromatase (P450arom) and Dmrt-1 complementary DNAs were isolated and their expression patterns were analyzed by competitive and conventional reverse transcriptase-polymerase chain reaction. While the P450arom gene was expressed predominantly in the ovary, Dmrt-1 was expressed exclusively in the testis. When larvae were reared at the female-producing temperature (28 degrees C) during the TSP, a strong expression of the P450arom gene and a complete suppression of the Dmrt-1 gene were induced in all experimental larvae. Up-regulation of the P450arom gene and down-regulation of the Dmrt-1 gene even in genetic males constitute a part of the molecular biological cascade for the temperature-dependent sex reversal from genetic males to phenotypic females in this salamander.
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PMID:Up-regulation of P450arom and down-regulation of Dmrt-1 genes in the temperature-dependent sex reversal from genetic males to phenotypic females in a salamander. 1650 70

Steroid hormones have important modulatory effects on the hair follicle, but the mechanisms by which they regulate human hair growth are still poorly understood. It is now clear that there are two distinct estrogen receptors (estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta)) that bind 17beta-estradiol. Since the follicular dermal papilla is known to control hair growth, and steroid hormones regulate receptor and aromatase expression in other tissues, we tested the hypothesis that steroid hormones would similarly modulate estrogen receptor and/or aromatase expression in cultured dermal papilla cells derived from human hair follicles. Primary cultures of non-balding occipital and frontal scalp and beard dermal papilla cells (n = 10) were established. Immunocytochemical studies showed the expression of ERalpha in both the cytoplasm and nucleus, whereas ERbeta was confined to the nuclei. The cells derived from occipital scalp were also incubated for 24 hours with 10 nM of either 17beta-estradiol, estrone, testosterone, 5alpha-dihydrotestosterone, 5alpha-androstane-3alpha, 17beta-diol, 5alpha-androstane-3beta, 17beta-diol, or 100 nM tamoxifen or dexamethasone in phenol red-free, serum-free medium to measure the steady-state levels of ERalpha, ERbeta, and aromatase mRNA by semiquantitative reverse transcriptase-PCR. Although androgens and estrogens did not alter ERalpha mRNA levels, treatment with dexamethasone significantly reduced ERalpha levels to 38% of the untreated control. By contrast, ERbeta mRNA levels were unaffected by any steroid treatment. Furthermore, dexamethasone significantly stimulated the expression of aromatase mRNA approximately 9-fold. Aromatase activity, assayed by the tritiated water method, was stimulated in both frontal scalp and beard dermal papilla cell cultures by dexamethasone. These observations provide evidence for a glucocorticoid-dependent mechanism whereby the selective action of estradiol via ERbeta may be promoted. Additionally, upregulation of aromatase combined with downregulation of ERalpha provides a basis for selective action of estradiol produced locally by autocrine or paracrine mechanisms.
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PMID:The modulation of aromatase and estrogen receptor alpha in cultured human dermal papilla cells by dexamethasone: a novel mechanism for selective action of estrogen via estrogen receptor beta? 1669 Nov 99

Large cell calcifying Sertoli cell tumors (LCCSCT) are associated with Carney complex and Peutz-Jeghers syndrome. The mechanisms linking these 2 genetic defects to the genesis of this tumor are obscure. Studies of CYP19 (aromatase) and transforming growth factor (TGF)-beta1 messenger RNA (mRNA) abundance, estrogen receptor (ER), TGFbeta1, and TGFbeta type II receptor (R) immunochemistry were carried out in the testis of a patient with this tumor to gain information on possible mechanisms of cell tumor development. Testicular tissue of a prepubertal patient, collected at gonadectomy, was separated into 2 macroscopically distinct fractions: tumoral nodules (Tu) and extratumoral, normal-looking testicular tissue (ExTu). The patient was a 9.5-year-old boy with a 5-year history of bilateral gynecomastia (Tanner stage 4), no pubic hair, incipient genital development, and bilateral testicular nodules. Multiple pigmented lesions of the skin were present. Bilateral mammectomy and gonadectomy was performed. RNA was extracted from Tu and ExTu for semiquantitative reverse transcriptase-polymerase chain reaction of CYP19 and TGFbeta1. Protein expression of ER, TGFbeta1, and TGFbeta type II R in Tu and ExTu was detected by immunohistochemistry. Cell proliferation was estimated by Ki-67 antigen immunochemistry and apoptosis using a modified TUNEL assay. Mean expression of aromatase and TGFbeta1 mRNAs in Tu was 6- and 2.3-fold higher than in ExTu, respectively (P<0.05). Tumoral cells exhibited ER staining with a predominant extranuclear localization. Positive staining of Sertoli cells in Tu was higher than in ExTu. TGFbeta1 immunostaining of the interstitial cells in Tu was higher than in ExTu. TGFbeta type II R immunostaining was detected in most Sertoli and interstitial cells, but intensity in ExTu was lower than in Tu. No significant difference was detected in the proliferation index, but in Tu, the percentage of Sertoli cells in apoptosis (1.4%) was significantly lower (P<0.01) than in ExTu (14.0%). The following hypothesis is proposed. The congenital gene defects of Carney complex or of Peutz-Jeghers syndrome might trigger a cascade of intracellular events that leads to overexpression of aromatase in Sertoli cells, favoring the development of a LCCSCT. At some point in the evolution of the disease, a mutational event might induce a higher expression of the ER. Also, TGFbeta1 protein expression is increased in neighboring cells. In this environment, TGFbeta1 might switch from tumor suppressor to oncogenic factor and, along with estrogen-ER complexes, might favor tumor progression by inhibiting apoptosis.
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PMID:High TGFbeta1, estrogen receptor, and aromatase gene expression in a large cell calcifying sertoli cell tumor (LCCSCT): implications for the mechanism of oncogenesis. 1694 77

The presence of a complex population of mRNAs in human mature spermatozoa is well documented; among them, transcripts of aromatase and ERs (oestrogen receptors) have been described but their significance is not clear. Therefore, to clarify the role of this complex population of mRNAs in human ejaculated sperm, we have isolated on discontinuous density gradients two main fractions from the same sample: high- and low-motile spermatozoa. The levels of different transcripts coding for molecules involved in nuclear condensation [Prm-1 (protamine 1) and Prm-2], capacitation [eNOS (endothelial nitric oxide synthase), nNOS (neuronal nitric oxide synthase), c-myc], motility and sperm survival (aromatase) have been assessed using semi-quantitative RT (reverse transcriptase)-PCR. The viability of sperm as well as the percentage of apoptosis were identical in high- and low-motile fractions. No significant change in the c-myc/Prm-2 ratio between the two populations of spermatozoa was observed. Conversely the amount of Prm-1 mRNA was significantly higher in low-motile than in high-motile fraction; in most of the high-motile sperm samples analysed, eNOS and nNOS transcripts were undetectable, whereas they were observed in low-motile sperm. Moreover, a partial or complete disappearance of c-myc transcripts was observed after capacitation. As to the aromatase expression, a significant decrease in the amount of transcripts in immotile sperm fraction was recorded in all samples studied. To conclude, analysing mRNA profiles in humans could be helpful either as a diagnostic tool to evaluate male fertility, since they reflect spermatogenesis gene expression, and/or a prognosis value for fertilization, since these RNAs are delivered to oocytes.
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PMID:RNA dynamics of fertile and infertile spermatozoa. 1751 68


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