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171.
Glucose Transporter (GLUT-4) Is Targeted to Secretory Granules in Rat Atrial Cardiomyocytes 总被引:6,自引:2,他引:4
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Jan W. Slot Gabriella Garruti Sally Martin Viola Oorschot George Posthuma Edward W. Kraegen Ross Laybutt Gatan Thibault David E. James 《The Journal of cell biology》1997,137(6):1243-1254
The insulin-responsive glucose transporter GLUT-4 is found in muscle and fat cells in the transGolgi reticulum (TGR) and in an intracellular tubulovesicular compartment, from where it undergoes insulindependent movement to the cell surface. To examine the relationship between these GLUT-4–containing compartments and the regulated secretory pathway we have localized GLUT-4 in atrial cardiomyocytes. This cell type secretes an antihypertensive hormone, referred to as the atrial natriuretic factor (ANF), in response to elevated blood pressure. We show that GLUT-4 is targeted in the atrial cell to the TGR and a tubulo-vesicular compartment, which is morphologically and functionally indistinguishable from the intracellular GLUT-4 compartment found in other types of myocytes and in fat cells, and in addition to the ANF secretory granules. Forming ANF granules are present throughout all Golgi cisternae but only become GLUT4 positive in the TGR. The inability of cyclohexamide treatment to effect the TGR localization of GLUT-4 indicates that GLUT-4 enters the ANF secretory granules at the TGR via the recycling pathway and not via the biosynthetic pathway. These data suggest that a large proportion of GLUT-4 must recycle via the TGR in insulin-sensitive cells. It will be important to determine if this is the pathway by which the insulin-regulatable tubulo-vesicular compartment is formed. 相似文献
172.
Carol A. Sheppard Peter B. Simpson Alan H. Sharp Frederick C. Nucifora Christopher A. Ross †G. David Lange James T. Russell 《Journal of neurochemistry》1997,68(6):2317-2327
Abstract: We have examined the mechanisms that underlie Ca2+ wave propagation in cultured cortical astrocytes. Norepinephrine evoked Ca2+ waves in astrocytes that began at discrete initiation loci and propagated throughout the cell by regenerative amplification at a number of cellular sites, as shown by very high Ca2+ release rates at these regions. We have hypothesized previously that domains displaying elevated Ca2+ release kinetics in astrocytes may correspond to sites of high inositol 1,4,5-trisphosphate receptor (InsP3 R) density. To examine this possibility, we compared the distribution pattern of endoplasmic reticulum (ER) and InsP3 Rs with Ca2+ release kinetics in subcellular regions during propagation of norepinephrine-evoked waves. 3,3'-Dihexyloxacarbocyanine iodide staining revealed that the ER in astrocytes exists as a meshwork of membranes extending throughout the cells, including fine processes. A specific antibody directed against type 2 InsP3 Rs (InsP3 R2) detected a 260-kDa band in western blotting of astrocyte membranes. Immunocytochemistry using this antibody stained the entire ER system in a punctate, variegated manner. When Ca2+ responses and InsP3 R2 immunofluorescence were compared in the same cell, domains of elevated Ca2+ response kinetics (high amplitude and rapid rate of rise) showed significant positive correlation with high local intensity of InsP3 R2 staining. It appears, therefore, that specializations in the ER responsible for discrete local Ca2+ release sites that support regenerative wave propagation include increased levels of InsP3 R2 expression. 相似文献
173.
Homogenates of specific brain regions of three sensory systems (auditory, olfactory, and visual) were prepared from pigmented Long-Evans Hooded rats and assayed for amino acid concentrations and activities of glutaminase, aspartate aminotransferase (total, cytosolic, and, by difference, mitochondrial), malate dehydrogenase, lactate dehydrogenase, and choline acetyltransferase. Comparing the quantitative distributions among regions revealed significant correlations between AAT and aspartate, between glutaminase and glutamate, between glutamate and glutamine, and between AAT plus glutaminase, or glutaminase alone, and the sum of aspartate, glutamate, and GABA, suggesting a metabolic pathway involving the synthesis of a glutamate pool as precursor to aspartate and GABA. Of the inhibitory transmitter amino acids, GABA concentrations routinely exceeded those of glycine, but glycine concentrations were relatively high in brainstem auditory structures. 相似文献
174.
G. M. Morris J. W. Hopewell G. A. Ross E. Whitehouse D. Wilding C. A. Scott 《Cell proliferation》1995,28(2):73-84
The daily oral administration of 3 ml of two oils (So-5407 and So-1129) containing essential fatty acids (EFAs) for 16 weeks resulted in a transient increase in cell proliferative activity in the skin of female Large White pigs. The So-5407 oil contained 7% gamma-linolenic acid (GLA) whereas So-1129 was an oil of similar composition, but with no GLA. Hyperplasia of the epidermis was observed after the administration of both oils, and this was characterized by an increase in the size of the rete pegs. The maximum effect occurred at 4 weeks after the start of oil administration, at which time the number of viable cell layers had increased by a factor of approximately 1.5, and mean epidermal thickness (excluding the stratum corneum) was approximately 40% greater than that of the epidermis prior to oil administration. There was a marked increase in the labelling index (LI) of the basal cell layer of the epidermis in pigs receiving So-5407. Maximum LIs were quantified at 4 weeks after the start of administration and were 18.8 ± 1.3% and 13.1 ± 1.7% for pigs receiving So-5407 and So-1129, respectively. After this time the LI declined prgressively and had returned to values within normal limits (P>0.1) by 8 weeks after the start of administration of both oils. A similar pattern of change in the LI was seen in the follicular epithelium, although the peak values at 4 weeks after the start of oil administration of 12.2 ± 1.8% and 10.8 ± 0.9 for the groups receiving So-5407 and So-1129, respectively, were lower than in the epidermis. Labelled cells were also counted in the papillary dermis and maximum values were again seen at 4 weeks after the start of oil administration. Of the two oils, So-1129 had the greatest effect, with the number of labelled cells in the papillary dermis being a factor of three to four-fold higher than in skin prior to oil administration, between 2 and 12 weeks after the start of administration. 相似文献
175.
Genetic regulation of gibberellin deactivation in Pisum 总被引:2,自引:0,他引:2
John J. Ross James B. Reid Stephen M. Swain Omar Hasan rew T. Poole Peter Hedden Christine L. Willis 《The Plant journal : for cell and molecular biology》1995,7(3):513-523
The regulation of gibberellin (GA) deactivation was examined using the sin (slender) mutation in the garden pea (Pisum sativum L.). This mutation blocks the deactivation of GA20, the precursor of the bioactive GA1. Firstly, crosses were made to combine sin with the GA biosynthesis mutations na, lhi and le-3. The combination sin na produced a novel phenotype, with long (‘slender’) basal internodes and extremely short (‘nana’) upper internodes. In contrast, the double mutant sin lhi was phenotypically dwarf. The mutation sin causes an accumulation of GA20 in maturing seeds, and this was unaffected by na, since the na mutation is not expressed in seeds. In contrast, lhi seeds did not accumulate GA20, since lhi imposes an early block on GA biosynthesis. Secondly, the effects of sin on several steps in GA deactivation were investigated. In maturing seeds, the mutation sin blocks two steps in GA20 metabolism, namely, GA20 to GA29, and GA29 to GA29-catabolite. In the vegetative plant, on the other hand, sin blocked the step GA20 to GA29, but not GA29 to GA29-catabolite; the steps GA20 to GA81 and GA20 to GA1 were also not impaired in this mutant. It is clear that the effects of sin, like those of na, are strongly organ-specific. The presence of separate enzymes for the steps GA20 to GA29 and GA29 to GA29-catabolite was suggested by the observation that GA8 inhibited the latter step, but not the former, and by the inability of GA20 and GA29 to inhibit each other's metabolism. It is suggested that the Sin gene may be a regulatory gene controlling the expression of two structural genes involved in GA deactivation. 相似文献
176.
Measurement of the adenine nucleotide and inorganic phosphate content of normoxic and ischaemic kidney in vivo has been made, comparing enzymic assay (after freeze-clamping and acid extraction) with quantification by 31P-n.m.r. Both methods give similar results for ATP, and n.m.r. quantification of Pi gives a value 25-50% of that obtained by enzymic assay. ADP, which is largely invisible to n.m.r. in the normoxic kidney, remains invisible during ischaemia despite a 2-3 fold rise in enzymically assayed ADP. N.m.r. and enzymic assay of the acid extracts give similar values for all metabolites measured. The question of ADP binding in the kidney is discussed, as are the implications for the metabolic regulation of ADP-dependent reactions. 相似文献
177.
178.
Subcellular localization of RNAs in transfected cells: role of sequences at the 5' terminus 总被引:4,自引:2,他引:2
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We have investigated the intracellular location of RNAs transcribed from transfected DNA. COS cells transfected with a clone containing the human adult beta globin gene contain three classes of globin RNAs. Their 3' termini and splice sites are indistinguishable from those of mature reticulocyte beta globin mRNA, and they are polyadenylated. However, as determined by S1 mapping, their 5' sequences are different. The 5' terminus of one is the same as that of mature beta globin mRNA (+1, cap site). The presumed 5' terminus of the second is located 30 nucleotides downstream from the cap site (+30). The third class contains additional nucleotides transcribed from sequences located 5' to the cap site (5' upstream RNA). The 5' upstream RNA molecules are restricted to the nucleus and are more stable than heterogeneous nuclear RNA. The +30 and +1 RNAs are located primarily in the cytoplasm. The data support the notion that nucleotide sequences and/or secondary modifications in the 5' region determine if an RNA is to be transported. 相似文献
179.
Summary Ammonia evolution and urease, phosphatase and protease activities were measured after field incubations of soil from 5 sites, differently influenced by penguin guano, from Cape Bird, Antarctica. Rates of NH3 production were low considering the large amounts of free NH3 in these soils. Enzyme activities were higher than expected and, under optimum conditions of temperature, moisture and substrate availability, were sufficient to degrade rapidly the accumulated organic matter in these soils. Laboratory incubations confirmed these high levels of activity; they also showed high dehydrogenase, but not sulphatase, activity. The surface horizon from the site that received continuous additions of fresh penguin excreta had considerably more enzyme activity than the other samples. The optimum activity of phosphatase in this sample was at pH 9.5-10. These enzyme activities are compared with those of New Zealand topsoils, and considered in relation to activities likely to occur under field conditions. Recommendations are made for further biochemical studies at Cape Bird. 相似文献
180.
J L Monro S Mollo S Brookbanks N Conway J K Ross 《BMJ (Clinical research ed.)》1978,1(6128):1684-1685
A study in Wessex has shown that at 1977 prices, and excluding the cost of equipment already installed in the unit, the cost of replacing a man''s aortic valve in this unit is about 1800 pounds. Nevertheless, this seems a small price to pay for return to health and full working capacity, particularly since such patients no longer need to draw social security benefits and their tax contributions will return to normal, thus probably paying for the operation within two years. 相似文献