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Query: EC:3.1.27.1 (
RNase
)
16,360
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The developmental regulation of rat brain-derived/Hep G2
glucose transporter
gene expression was studied by means of Northern blot hybridization, using a rat brain
glucose transporter
cDNA probe, in order to directly quantify steady state
glucose transporter
mRNA levels. The results obtained showed different tissue-specific patterns of
glucose transporter
mRNA levels during ontogenesis; while in brain there was a sustained increase in the levels of the message from 20 days embryogenesis until 50 days postnatal, other organs such as heart, lung, liver, and muscle expressed maximal levels of the
glucose transporter
mRNA in 20-day fetuses and 1-day neonates, decreasing subsequently to very low levels. The relative expression of the
glucose transporter
mRNA in the different tissues, at both fetal and adult stages, was analyzed using a solution hybridization-
RNase
protection assay. This approach revealed that, while the heart expresses the highest levels of
glucose transporter
mRNA at 20 days of fetal life, the brain shows the highest levels at the adult stage. These results indicate a tissue-specific ontogenic pattern of
glucose transporter
gene expression, suggesting a developmental role for this
glucose transporter
gene product.
...
PMID:Developmental regulation of rat brain/Hep G2 glucose transporter gene expression. 271 Jan 34
Differential developmental regulation of pancreas-specific genes has not been reported for the human fetal pancreas. We have therefore undertaken a systematic, quantitative analysis of the transcriptional levels of various genes in the human pancreas at different stages of fetal and postnatal development. Using sensitive
ribonuclease
protection assays, in situ hybridization, and the polymerase chain reaction, our results indicate the following: 1) Transcriptional levels of insulin and amylin remain lower in the fetal than in the adult pancreas, whereas glucagon and somatostatin mRNA levels are consistently greater after 14 wk gestation than postnatally. These results are in agreement with previous immunohistochemical studies of these gene products. 2) The reg gene exhibits a 20-fold increase in mRNA levels after 16 wk gestation. The gene is expressed exclusively in the acinar cells and does not colocalize with insulin. This restricted exocrine expression does not indicate a direct role for the reg gene in islet development. 3) Glucose transporter 2 and glucokinase mRNA are detectable as early as 13 wk gestation and remain low throughout development. Glucose transporter 1 reaches adult transcriptional levels by 18 wk gestation. The early detection of
glucose transporter
2 and glucokinase implies that lack of expression of these "glucose sensor" genes does not account for the known insensitivity of the fetal beta-cells to glucose.
...
PMID:Developmental gene expression in the human fetal pancreas. 752 96
To examine the mechanisms responsible for tissue-specific, nutritional, and metabolic regulation of the GLUT4/muscle-adipose specific
glucose transporter
, we isolated and characterized the properties of the rat GLUT4 gene. Examination of the sequenced 2.5-kilobase flanking DNA revealed substantial identity with that of the mouse and human GLUT4 genes, with the greatest degree of sequence identity within the proximal 1000 basepairs up-stream of the GLUT4 open reading frame. Primer extension analysis identified a unique single transcription initiation site 176 basepairs up-stream from the start of translation. However,
ribonuclease
mapping revealed the presence of a previously undescribed alternatively spliced form of GLUT4 messenger RNA. Approximately 75% of the GLUT4 transcripts consisted of a fully spliced messenger RNA, and 25% was expressed as an unspliced intron-containing species. The ratios of 5' spliced and unspliced messages were invariant in adipose, cardiac, and skeletal muscle tissues. In vitro translation of reporter constructs containing both the spliced and unspliced leader demonstrated a functional difference between these two transcripts, with the unspliced form translated approximately 5-fold more than the fully spliced species. These data demonstrate the presence of 5'-heterogeneity of the GLUT4 transcripts, which underlies differences in translational efficiency in vitro.
...
PMID:Characterization of 5'-heterogeneity of the rat GLUT4/muscle-adipose glucose transporter gene product. 772 Jun 44
Previous studies have indicated that insulin secretion in response to glucose diminishes with age but insulin synthesis and gene transcription do not. To determine whether expression of genes other than those that encode insulin are subject to age-related changes that could alter pancreatic islet function, mRNAs for insulins I and II, amylin,
glucose transporter
2 (GluT2), glucagon, and glucokinase were quantified in 2-, 6-, 12-, and 24-month-old Fischer 344 rats using species-specific
ribonuclease
(
RNase
) protection assays. There was only a modest (1.2- to 1.3-fold) increase in insulin I and insulin II mRNAs between ages 2 and 12 months. There were no statistically significant changes in levels of glucokinase mRNA with age. In contrast, the abundances of amylin, GluT2, and glucagon mRNAs all doubled during the same period. Variance in values from 24-month-old rats was too great to allow conclusions, except that the ratio of insulin II mRNA to insulin I mRNA increased with age. This change was not related to islet mass or total insulin mRNA abundance because it persisted at age 24 months, when total mRNA abundance had decreased. These results indicate that aging is associated with significant alterations in the relative proportion of expression of pancreatic islet cell genes implicated in insulin secretion and in intraislet glucose metabolism.
...
PMID:Age-related changes in pancreatic islet cell gene expression. 788 76
In pancreatic beta-cells, the high Km
glucose transporter
GLUT2 catalyzes the first step in glucose-induced insulin secretion by glucose uptake. Expression of the transporter has been reported to be modulated by glucose either at the protein or mRNA levels. In this study we used the differentiated insulinoma cell line INS-1 which expresses high levels of GLUT2 and show that the expression of GLUT2 is regulated by glucose at the transcriptional level. By run-on transcription assays we showed that glucose induced GLUT2 gene transcription 3-4-fold in INS-1 cells which was paralleled by a 1.7-2.3-fold increase in cytoplasmic GLUT2 mRNA levels. To determine whether glucose regulatory sequences were present in the promoter region of GLUT2, we cloned and characterized a 1.4-kilobase region of mouse genomic DNA located 5' of the translation initiation site. By
RNase
protection assays and primer extension, we determined that multiple transcription initiation sites were present at positions -55, -64, and -115 from the first coding ATG and which were identified in liver, intestine, kidney, and beta-cells mRNAs. Plasmids were constructed with the mouse promoter region linked to the reporter gene chloramphenicol acetyltransferase (CAT), and transiently and stably transfected in the INS-1 cells. Glucose induced a concentration-dependent increase in CAT activity which reached a maximum of 3.6-fold at 20 mM glucose. Similar CAT constructs made of the human GLUT2 promoter region and the CAT gene displayed the same glucose-dependent increase in transcriptional activity when transfected into INS-1 cells. Comparison of the mouse and human promoter regions revealed sequence identity restricted to a few stretches of sequences which suggests that the glucose responsive element(s) may be conserved in these common sequences.
...
PMID:Characterization of the murine high Km glucose transporter GLUT2 gene and its transcriptional regulation by glucose in a differentiated insulin-secreting cell line. 792 31
A 246-bp fragment of porcine glucose transporter 4 (GLUT4) cDNA was cloned by polymerase chain reaction (PCR) from porcine adipose tissue RNA. Nucleotide sequences 1-138 and 139-246 of the GLUT4 cDNA share 78% sequence identity with exon 4a and 91% sequence identity with exon 4b of the human GLUT4 gene, respectively. The GLUT4 cDNA fragment was subcloned into pGEM-4Z vector to synthesize a highly specific riboprobe that hybridized only to human GLUT4 cDNA but not to human
glucose transporter
1 (GLUT1) cDNA. Northern blot analysis of total RNA revealed the presence of a single transcript of 2.8 kb in porcine adipose tissue. Cloning a fragment of the GLUT4 cDNA enabled us to develop a
ribonuclease
protection assay for detecting porcine GLUT4 mRNA. The
ribonuclease
(
RNase
) protection assay is highly reproducible and retains a sensitivity level to as little as 2 pg of GLUT4 mRNA. The standard curve was linear between 2 and 128 pg of sense-strand GLUT4 RNA (r = .994). The ability to detect small quantities of GLUT4 mRNA is important when the abundance of GLUT4 mRNA is low and the quantity of tissue is limiting (e.g., when RNA is extracted from cultured adipose tissue). When porcine adipose tissue explants were cultured in the presence of insulin (10 ng/mL), GLUT4 mRNA abundance was increased. Development of a sensitive assay to quantify GLUT4 mRNA in porcine adipose tissue will enable us to conduct studies to increase our understanding of the molecular mechanisms by which porcine somatotropin (pST) regulates GLUT4 gene expression.
...
PMID:Cloning of a pig glucose transporter 4 cDNA fragment: use in developing a sensitive ribonuclease protection assay for quantifying low-abundance glucose transporter 4 mRNA in porcine adipose tissue. 805 64
The effects of fasting and refeeding on the levels of mRNA encoding the insulin-sensitive
glucose transporter
(GLUT4) in rat cardiac and skeletal muscle were investigated using solution hybridization/
RNase
protection assays with a rat GLUT4 antisense RNA probe. In addition, the effects of these nutritional states on GLUT1 mRNA levels in several non-insulin-sensitive tissues were examined using a GLUT1 antisense RNA probe. Fasting for 48 hr significantly decreased GLUT4 mRNA levels in heart, with levels significantly increased over control levels by 24 hr after refeeding. In contrast, GLUT4 mRNA levels in skeletal muscle increased with fasting and returned to control levels with refeeding. No significant changes in GLUT1 mRNA were seen after fasting and refeeding in several non-insulin-sensitive tissues studied. These results suggest that altered GLUT4 gene expression is observed in different nutritional (insulin) states in insulin-sensitive tissues, and suggests a potential role for insulin in mediating these changes in gene expression.
...
PMID:Nutritional regulation of insulin-sensitive glucose transporter gene expression in rat cardiac muscle. 850 58
The murine facilitative
glucose transporter
isoform 3 (Glut 3) is developmentally regulated and is predominantly expressed in neurons and trophoblasts. Employing the primer extension and
RNase
protection assays, the transcription start site (denoted as +1) of the murine Glut 3 gene was localized to 305 base pairs (bp) 5' to the ATG translation start codon. Transient transfection assays in N2A, H19-7 neuroblasts, and HRP.1 trophoblasts using sequential 5'-deletions of the murine Glut 3-luciferase fusion gene indicated that the -203 to +237 bp region with reference to the transcriptional start site contained promoter activity. Repressor function was limited to the -137 to -130 bp region within the transcriptional activation domain. The nuclear factors Sp1 and Sp3 bound this GC-rich region in N2A, H19-7, and HRP.1 cells. Dephosphorylation of Sp1 was essential for Glut 3 DNA binding. The related Sp3 protein also bound this same region of mouse Glut 3 in all three cell lines. Mutations of the Sp1-binding site employed in transient transfection and mobility shift assays confirmed the nature of the DNA-binding proteins, while supershift assays with anti-Sp1 and anti-Sp3 IgGs characterized the differences in the two DNA-binding proteins. Co-transfection of the Glut 3-luciferase fusion gene with or without mutations of the Sp1-binding site along with the Sp1 or Sp3 expression vectors in Drosophila SL2 cells confirmed a reciprocal effect, with Sp1 suppressing and Sp3 activating Glut 3 gene transcription.
...
PMID:Sp1 and Sp3 regulate transcriptional activity of the facilitative glucose transporter isoform-3 gene in mammalian neuroblasts and trophoblasts. 976 77
Depletion of GLUT4, the primary
glucose transporter
protein in adipose tissue and skeletal muscle, is reported to contribute to insulin resistance in pregnancy or diabetes. To examine this phenomenon, the expression of GLUT4 protein was assessed by Western blotting in streptozotocin-induced diabetic pregnant rats. In adipose tissue, relative to control, it was decreased by 30% in the normal pregnant group (p<0.001), by 37% in the diabetic nonpregnant group (p<0.01) and by 65% in the diabetic pregnant group (p<0.001). On the other hand, no significant variation was evident among the groups in skeletal muscle. To assess the mechanisms responsible for depletion of GLUT4 protein in adipose tissue, we quantitated levels of GLUT4 mRNA with a
RNase
protection assay. It was decreased by 44% in the normal pregnant group (p<0.05) and by 55% in the diabetic pregnant group (p<0.05), but not altered in the diabetic nonpregnant group. These results suggest that the depletion of GLUT4 protein in adipose tissue is a factor contributing to insulin resistance in pregnancy or diabetes, especially when the two states exist in combination.
...
PMID:Expression of GLUT4 glucose transporter protein in adipose tissue and skeletal muscle from streptozotocin-induced diabetic pregnant rats. 1056 52
Regulation of endometrial glucose transport is important for the decidualization process. Therefore, we have examined the expression of the
glucose transporter
protein isoform 1 (GLUT1) in endometrial samples during the menstrual cycle and in decidual tissue by immunohistochemistry, and GLUT1 mRNA by
RNase
protection assays. GLUT1 protein was not detected in proliferative endometrial samples, but was highly expressed in decidual tissue. Placental tissue was highly positive. GLUT1 mRNA could be detected in endometrial samples with an increase in endometria of the late secretory phase (day 25-28) and maximum concentration in the decidua of the 9th-10th week of gestation. Our results show that GLUT1 is differentially expressed in the different phases of the human endometrium with a maximum in the human decidua. Therefore, GLUT1 may be an important marker for endometrial differentiation.
...
PMID:Expression of glucose transporter 1 in human endometrial and decidual tissue. 1144 34
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