Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.26.9 (ribonuclease)
 6,589 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Our laboratory and others have reported that treatment of GT1-7 cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), inhibits transcription of the pro-GnRH gene and decreases messenger RNA (mRNA) levels. We were interested in whether translation of the existing GnRH mRNA decreases in parallel with these other indexes of biosynthesis after PMA treatment. GT1-7 cells were treated with PMA (100 nM) or vehicle for 0, 1, or 4 h. The cytosolic ribosome-associated RNA was isolated and layered on a continuous (10-40%) sucrose gradient, and fractions were analyzed for the distribution of ribosome-associated GnRH mRNA through the gradient by ribonuclease protection assay. The mRNA found in the lighter fractions is associated with fewer ribosomes per RNA, suggesting that these fractions are translated less efficiently, and RNA recovered from heavier fractions has a higher number of ribosomes per mRNA, representing mRNA that is more actively translated. We found that the distribution of the ribosome-associated GnRH mRNA was shifted into lighter fractions (i.e. fewer ribosomes per mRNA) after PMA treatment, indicating that a decrease in the translational efficiency of GnRH mRNA occurs after PMA treatment. Thus, PMA exerts inhibitory effects on translation of GnRH mRNA as well as on gene transcription, mRNA stability, and mRNA levels.
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PMID:Translational efficiency of gonadotropin-releasing hormone messenger ribonucleic acid is negatively regulated by phorbol ester in GT1-7 cells. 789 72

The immortalized GT1-7 cell line synthesizes and secretes GnRH, the key hormone of reproduction. However, GT1-7 cells lack the normal inputs from neurotransmitters, growth factors, and steroids, which are involved in the maturation and maintenance of GnRH neurons in the brain. We examined the effects of the neurotrophic factor insulin-like growth factor-I (IGF-I) on GnRH gene expression and the mechanism for these changes. Initially, effects of IGF-I on GnRH gene expression were determined by ribonuclease protection assay. In time-course experiments, IGF-I treatment caused significant increases in nuclear GnRH primary transcript levels, an index of GnRH gene transcription, 4 and 8 h after initiation of IGF-I treatment. GnRH messenger RNA (mRNA) levels in the cytoplasm were stimulated by IGF-I at 24 h of treatment. IGF-I also affected GT1-7 cell morphology, with an increase in process extension and cell-cell contacts. In contrast, GnRH peptide levels in the medium were initially stimulated and then suppressed by IGF-I, indicating an uncoupling of biosynthesis and secretion. The increase in GnRH mRNA levels induced by IGF-I is probably caused by a transcriptional mechanism, as evidenced by the increase in GnRH primary transcript levels before a change in GnRH mRNA levels, as well as our finding of a similar GnRH mRNA half-life for both control and IGF-I-treated cells. Interestingly, GT1-7 cells themselves were observed to express IGF-I immunoreactivity, suggesting the possibility of autoregulation by this neurotrophic factor. It is concluded that IGF-I is an important modulator of GnRH gene expression and release in the GT1-7 cell line. The reported stimulatory effects of IGF-I in vivo, and its hypothesized role in the development of GnRH neurons in the brain, suggest that IGF-I may make the GT1-7 cells line more like a mature GnRH neuron, as a model for future studies.
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PMID:Insulin-like growth factor-I effects on gonadotropin-releasing hormone biosynthesis in GT1-7 cells. 949 46

The role of calcium in the regulation of GnRH gene expression and the mechanism for its effects were examined in the present study. Using the immortalized hypothalamic GT1-7 cell line, which synthesizes and secretes GnRH, we demonstrated by ribonuclease protection assay and Northern blot analysis that these cells respond to treatment with the calcium ionophores ionomycin and A23187 with an inhibition of transcription of the GnRH gene and decreases in GnRH messenger RNA (mRNA) levels. Ionomycin treatment caused the GnRH mRNA half-life to decrease from 25 to 9 h, concomitant with a decrease in mRNA poly(A) tail length, suggesting that ionomycin causes a decrease in GnRH mRNA stability. The ionomycin inhibitory effect on GnRH cytoplasmic mRNA levels was significantly inhibited in the presence of cycloheximide or the RNA synthesis inhibitor 5,6-dichloro-1beta-ribofuranosylbenzimidazole, indicating that novel protein/RNA synthesis is obligatory for this effect. We conclude that an increase in calcium levels caused by ionomycin inhibits GnRH gene expression at multiple levels, including GnRH gene transcription and mRNA stability in GT1-7 cells.
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PMID:The role of calcium in the transcriptional and posttranscriptional regulation of the gonadotropin-releasing hormone gene in GT1-7 cells. 960 73

A neurally expressed heterotrimeric G protein beta subunit, Gbeta(5), has been found to exhibit functional specialization with respect to its interactions with effector targets and Galpha subunits. A splice variant of Gbeta(5) that contains an N-terminal 42-residue extension, Gbeta(5)-long, has been described in the retina. To define better the potential range of its specialized interactions, analysis of Gbeta(5) gene transcript and protein expression in mouse brain and other tissues and cell lines was performed. Quantification by ribonuclease protection assay of Gbeta(5) transcript expression in the developing brain demonstrates a fivefold increase that occurs postnatally. Analysis of transcript expression by in situ hybridization and ribonuclease protection assay indicates that the Gbeta(5) gene is differentially expressed among multiple adult mouse brain regions, including the motor and occipital cortex, the olfactory bulb and associated rhinencephalic structures, hypothalamus, pontine cochlear nuclei, and Purkinje cells in the cerebellum. Gbeta(5) is also expressed in several cultured cell lines of neuroendocrine origin, including murine alphaT3-1 pituitary gonadotrophs and GT1-7 hypothalamic cells, and rat PC12 pheochromocytoma cells. Immunoblotting of tissue homogenates with antibodies to two peptides common to Gbeta(5) and Gbeta(5)-long confirmed expression of Gbeta(5) in multiple brain regions and in spinal cord and expression of Gbeta(5)-long in retina. Taken together, these results suggest that the specialized molecular properties of Gbeta(5) have been adapted to diverse neural functions in the adult brain.
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PMID:Differential expression of the G protein beta(5) gene: analysis of mouse brain, peripheral tissues, and cultured cell lines. 1085 85

Regulation of extracellular excitotoxins by glial and neuronal glutamate transporters is critical to maintain synaptic terminal integrity. Factors interfering with the normal functioning of these transporters might be involved in neurodegeneration. Among them, recent studies have shown that hypoxia alters glutamate transporter function; however, it is unclear if hypoxia has an effect on the expression of glutamate transporters and which intracellular signaling pathways are involved. The C6 rat glial and GT1--7 mouse neuronal cell lines were exposed to hypoxic conditions (5% CO(2), 95% N(2)) and levels of glutamate transporter mRNA were determined by ribonuclease protection assay. After 21 hr, there was a 100% increase in levels of rat excitatory amino acid transporter 3 (EAAT3) mRNA in C6 cells and a 600% increase in levels of murine EAAT2 mRNA in GT1--7 cells. There was a similar increase in mRNA levels after hypoxia in C6 cells transfected with human EAAT2, whereas reoxygenation normalized the expression levels of glutamate transporters. Although the expression of EAATs was associated with increased immunoreactivity by Western blot, functioning of the transporters was decreased as evidenced by D-aspartate uptake. Finally, although the protein kinase C stimulator phorbol-12-myristate-13-acetate enhanced EAAT2 mRNA levels after hypoxia, protein kinase C inhibitor bisindolylmaleimide I had the opposite effect. Taken together, this study suggests that the hypoxia is capable of upregulating levels of EAATs via a protein kinase C-dependent compensatory mechanism. This increased expression is not sufficient to overcome the decreased functioning of the EAATs associated with decreased ATP production and mitochondrial dysfunction.
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PMID:Altered expression of glutamate transporters under hypoxic conditions in vitro. 1128 47