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1.
Aquaporins in complex tissues. I.Developmental patterns in respiratory and glandular tissues of rat 总被引:14,自引:0,他引:14
King Landon S.; Nielsen Soren; Agre Peter 《American journal of physiology. Cell physiology》1997,273(5):C1541
Developmentalexpression of aquaporin water transport proteins is not well understoodin respiratory tract or secretory glands; here we define aquaporinprotein ontogeny in rat. Expression of aquaporin-3 (AQP3), AQP4, andAQP5 proteins occurs within 2 wk after birth, whereas AQP1 firstappears before birth. In most tissues, aquaporin protein expressionincreases progressively, although transient high-level expression isnoted in distal lung (AQP4 at postnatal day+2) and trachea (AQP5 at postnatalday +21 and AQP3 at postnatal day+42). In mature animals, AQP5 is abundant in distallung and salivary glands, AQP3 and AQP4 are present in trachea, andAQP1 is present in all of these tissues except salivary glands.Surprisingly, all four aquaporin proteins are highly abundant innasopharynx. Unlike AQP1, corticosteroids did not induce expression ofAQP3, AQP4, or AQP5 in lung. Our results seemingly implicate aquaporinsin proximal airway humidification, glandular secretion, and perinatalclearance of fluid from distal airways. However, the studies underscorea need for detailed immunohistochemical characterizations anddefinitive functional studies. 相似文献
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Aquaporins in complex tissues: distribution of aquaporins 1-5 in human and rat eye 总被引:11,自引:0,他引:11
Hamann Steffen; Zeuthen Thomas; Cour Morten La; Nagelhus Erlend A.; Ottersen Ole Petter; Agre Peter; Nielsen Soren 《American journal of physiology. Cell physiology》1998,274(5):C1332
Multiple physiological fluid movements areinvolved in vision. Here we define the cellular and subcellular sitesof aquaporin (AQP) water transport proteins in human and rat eyes byimmunoblotting, high-resolution immunocytochemistry, and immunoelectronmicroscopy. AQP3 is abundant in bulbar conjunctival epithelium andglands but is only weakly present in corneal epithelium. In contrast, AQP5 is prominent in corneal epithelium and apical membranes of lacrimal acini. AQP1 is heavily expressed in scleral fibroblasts, corneal endothelium and keratocytes, and endothelium covering thetrabecular meshwork and Schlemm's canal. Although AQP1 is plentiful inciliary nonpigmented epithelium, it is not present in ciliary pigmentedepithelium. Posterior and anterior epithelium of the iris and anteriorlens epithelium also contain significant amounts of AQP1, but AQP0(major intrinsic protein of the lens) is expressed in lens fiber cells.Retinal Müller cells and astrocytes exhibit notableconcentrations of AQP4, whereas neurons and retinal pigment epitheliumdo not display aquaporin immunolabeling. These studies demonstrateselective expression of AQP1, AQP3, AQP4, and AQP5 in distinct ocularepithelia, predicting specific roles for each in the complex networkthrough which water movements occur in the eye. 相似文献
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Subcellular distribution and activity in different rat tissues of a deoxyinosine-activated nucleotidase.
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A nucleotidase (EC 3.1.3.31) isolated previously from rat liver cytosol was specifically measured in 14 different rat tissues and in subcellular fractions of liver and spleen, taking advantage of the stimulation exerted on it by deoxyinosine. The intracellular distribution studies showed that the enzyme is located almost entirely in the soluble cytoplasm except for the possible presence of 1-2% of the enzyme in the nucleus. The enzyme was present in various amounts in all the tissues studied. Spleen, thymus, and intestinal mucosa showed higher specific activities than any other tissue. On a per cell basis spleen, liver and intestinal mucosa had the highest enzyme activity, whereas bone marrow, brain, thymus, heart and skeletal muscle had activities in the lower range. The results may suggest that the enzyme plays a role in the recovery of endogenous nuclear material for nucleic acid synthesis. 相似文献
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In rat cerebellum the major portion of guanylate cyclase was found to be particulate-bound. The properties of particulate and supernatant guanylate cyclases from the cerebellum were comparatively examined. Both enzymes required the same optimal concentration of Mn2+ and were stimulated by Ca2+ in the presence of a low concentration of Mn2+. But dispersion of the particulate enzyme with Triton X-100 altered the Mn2+ concentration producing maximum activity and the inhibitory effect of Ca2+. The subcellular distributions of guanylate and adenylate cyclases were also studied in rat cerebellum. The major portions of the two cyclases were found in the mitochondrial fraction. The submitochondrial fractions separated by sucrose gradient showed that the major activities of both cyclases were concentrated in the fraction containing mainly nerve ending particles. 相似文献
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P L Sannes M Eguchi S S Spicer 《The journal of histochemistry and cytochemistry》1979,27(8):1180-1181
Methods for visualization of complex carbohydrates ultrastructurally were employed to study specific organelles of the rat monocyte and macrophage. Vicinal glycols of glycoconjugates were demonstrated with the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) postembedding sequence and acid groups were delineated by the dialyzed iron (DI) and high iron diamine (HID) preembedding techniques. Lysosomal bodies were generally found reactive with all three methods, although those of monocytes from the bone marrow and peripheral blood were notably lacking in acidic groups. The Golgi complex was consistently PA-TCH-SP-reactive, as were associated vesicles and occasional cisternal expansions, possibly related to GERL. Numerous cytoplasmic vesicles and small granulated structures and cisternae of the rough endoplasmic reticulum were also PA-TCH-SP-reactive. 相似文献
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Subcellular distribution and movement of 5''-nucleotidase in rat cells. 总被引:33,自引:16,他引:17
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1. Cell-surface 5'-nucleotidase was assayed by incubating whole-cell suspensions with 5'[3H]-AMP in iso-osmotic buffer and measuring [3H]adenosine production. The activity of cell-surface 5'-nucleotidase in hepatocytes, adipocytes and lymphocytes isolated from the rat was 15.0, 0.5 and 0.8pmol/min per cell at 37 degrees C respectively. 2. Disruption of the cells by vigorous mechanical homogenization or detergent treatment exposed additional 5'-nucleotidase activity, which represented 52%, 25% and 21% of the total activity in the three cell types respectively. This increase in 5'-nucleotidase activity which occurred when the cells were homogenized was due to a second pool of 5'-nucleotidase within the cell, rather than activation of the cell-surface enzyme. 3. In hepatocytes the intracellular 5'-nucleotidase activity was membrane-bound, indistinguishable from cell-surface 5'-nucleotidase in its inhibition by rabbit anti-(rat liver 5'-nucleotidase) serum and its kinetics with AMP, and was located on the extracytoplasmic face of vesicles within the cell. 4. The cell-surface 5'-nucleotidase of rat hepatocytes was rapidly inhibited when rabbit anti-(rat liver 5'-nucleotidase) serum or concanavalin A was added to the medium at 37 degrees C. Incubation with antiserum for 5 min at 37 degrees C inhibited 83 +/- 3% of the cell-surface enzyme. 5. Incubation of hepatocytes with exogenous antiserum or concanavalin A for 30 min at 37 degrees C resulted in over 50% inhibition of the intracellular enzyme. This inhibition was not prevented by disruption of the cytoskeleton or by ATP depletion. 6. Incubation of hepatocytes with exogenous antiserum or concanavalin A for up to 2h at 0 degrees C caused little or no inhibition of the intracellular enzyme, but over 75% inhibition of the cell-surface enzyme. 7. When surface-inhibited hepatocytes were washed and resuspended in buffer at 37 degrees C, 5'-nucleotidase was observed to redistribute from the intracellular pool to the cell surface. 相似文献
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Subcellular distribution and characteristics of trihydroxycoprostanoyl-CoA synthetase in rat liver. 总被引:2,自引:0,他引:2
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L Schepers M Casteels K Verheyden G Parmentier S Asselberghs H J Eyssen G P Mannaerts 《The Biochemical journal》1989,257(1):221-229
The subcellular distribution and characteristics of trihydroxycoprostanoyl-CoA synthetase were studied in rat liver and were compared with those of palmitoyl-CoA synthetase and choloyl-CoA synthetase. Trihydroxycoprostanoyl-CoA synthetase and choloyl-CoA synthetase were localized almost completely in the endoplasmic reticulum. A quantitatively insignificant part of trihydroxycoprostanoyl-CoA synthetase was perhaps present in mitochondria. Peroxisomes, which convert trihydroxycoprostanoyl-CoA into choloyl-CoA, were devoid of trihydroxycoprostanoyl-CoA synthetase. As already known, palmitoyl-CoA synthetase was distributed among mitochondria, peroxisomes and endoplasmic reticulum. Substrate- and cofactor- (ATP, CoASH) dependence of the three synthesis activities were also studied. Cholic acid and trihydroxycoprostanic acid did not inhibit palmitoyl-CoA synthetase; palmitate inhibited the other synthetases non-competitively. Likewise, cholic acid inhibited trihydroxycoprostanic acid activation non-competitively and vice versa. The pH curves of the synthetases did not coincide. Triton X-100 affected the activity of each of the synthetases differently. Trihydroxycoprostanoyl-CoA synthetase was less sensitive towards inhibition by pyrophosphate than choloyl-CoA synthetase. The synthetases could not be solubilized from microsomal membranes by treatment with 1 M-NaCl, but could be solubilized with Triton X-100 or Triton X-100 plus NaCl. The detergent-solubilized trihydroxycoprostanoyl-CoA synthetase could be separated from the solubilized choloyl-CoA synthetase and palmitoyl-CoA synthetase by affinity chromatograpy on Sepharose to which trihydroxycoprostanic acid was bound. Choloyl-CoA synthetase and trihydroxycoprostanoyl-CoA synthetase could not be detected in homogenates from kidney or intestinal mucosa. The results indicate that long-chain fatty acids, cholic acid and trihydroxycoprostanic acid are activated by three separate enzymes. 相似文献
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Lysophosphatidylinositol acyltransferase utilizing arachidonoyl CoA and lysophosphatidylcholine acyltransferase utilizing linoleoyl CoA were measured in subcellular fractions of rat brain. In general, the distribution of activities paralleled that of NADPH--cytochrome c reductase. It is concluded that the endoplasmic reticulum is the major site of these acyltransferase activities in rat brain. 相似文献
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Farnesyl protein transferase (FPT) activity was measured in rat liver subcellular fractions by using an unspecific acceptor for the farnesyl groups. The highest specific activity was found in mitochondria and it exceeded that of the microsomes three-fold. Considerably lower specific activities were found in the nuclei and cytosol. Further subfractionation revealed that the mitochondrial FPT activity is located in the matrix. The beta-subunit of the mitochondrial enzyme has an apparent molecular mass of 46 kDa, which is similar to its cytosolic counterpart. The results suggest that protein farnesylation can take place in a number of subcellular organelles. 相似文献
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The subcellular distribution of rat liver porin was investigated using the immunoblotting technique and monospecific antisera against the protein isolated from the outer membrane of rat liver mitochondria. Subfractionation of mitochondria into inner membranes, outer membranes and matrix fractions revealed the presence of porin only in the outer membranes. Porin was also not detected in highly purified subcellular fractions, including plasma membranes, nuclear membranes, Golgi I and Golgi II, microsomes and lysosomes. Thus, liver porin is located exclusively in the outer mitochondrial membrane. 相似文献
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—Certain of the sulphur containing amino acids have been associated with synaptic transmission in the central nervous system. The enzymes involved in the synthesis of these putative neurotransmitter or modulator compounds have a different subcellular distribution in rat brain from those enzymes that catalyse the synthesis of other compounds in this pathway. Methionine adenosyltransferase and 5-methyltetrahydrofolate-homocysteine methyltransferase catalyse reactions that maintain the methylation functions of the pathway and are found in soluble fractions. Cystathionine β-synthase, cystathionase, cysteine dioxygenase and cysteine sulphinic acid decarboxylase catalyse the synthesis of those sulphur-containing amino acids implicated in neurotransmitter functions and these enzymes have both paniculate and soluble components. Serine hydroxymethyltransferase, which also has a particulate fraction in brain, is responsible for the synthesis of the neurotransmitter glycine, in addition to its role in the methionine-related metabolism of folate. 相似文献
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T Noguchi M Nakatani Y Minatogawa M Morimoto R Kido 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1975,356(8):1245-1250
Kynurenine transaminase activity in rat kidney was found in both the mitochondrial and supernatant fractions. These fractions contained (a) kynurenine pyruvate transaminase, which showed a preference for pyruvate as amino acceptor, and had a pH optimum between 8.0 and 8.5, and (b) kynurenine 2-oxoglutarate transaminase, with a preference for 2-oxoglutarate and a pH optimum between 6.0 and 6.5. The apparent Km value of the former enzyme for L-kynurenine was much lower than that of the latter enzyme. 相似文献
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The subcellular distribution of leucine- and methionine-enkephalin in rat brain was studied using a highly selective and sensitive radioimmunoassay. About 85% of the total recoverable activity of each peptide was present in crude synaptosomal and microsomal fractions which contained about 60% and 25% respectively. Total opioid activity in brain subcellular extracts was measured by competition for opiat receptor binding. It is concluded that enkephalin accounts for the majority of the opioid activity in the brain extracts. It seems unlikely that the enkephalin in microsomal fractions are exclusively associated with opiate receptors present in these fractions. 相似文献