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Query: UNIPROT:P56851 (epididymal)
 11,273 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Calcium accumulation by ram sperm in the presence of 3.0 microM ionophore A23187 increased greatly when the pH of the medium was raised from 7.5 to 8.5. The increase in calcium uptake was remarkable between pH 7.5 and 8.0 and was paralleled by increased oxygen consumption. There was extensive vesiculation of the plasma membrane and membranes of the acrosome and postnuclear cap of ram sperm incubated with the ionophore and calcium at pH 8.0. The mitochondria of the midpiece contained pale and expanded cristae similar to ATP-deprived mitochondria in the "condensed" configuration. Such changes were not observed in ram sperm incubated under similar conditions at pH 7.0. The ionophore-induced calcium and oxygen uptake of cauda and caput epididymal boar sperm also increased with increased pH, but the effect commenced at a lower pH. Control cauda epididymal boar sperm (i.e., without ionophore) had higher oxygen uptake, glucose breakdown, and lactic acid production than caput sperm. The influence of pH on the operation of the membrane calcium pumps between ram boar sperm indicates species differences.
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PMID:Calcium uptake, respiration, and ultrastructure of sperm exposed to ionophore A23187. 312 77

1. Brown adipocytes were isolated from the interscapular depot of male rats maintained at approx. 21 degrees C. In some experiments parallel studies were made with white adipocytes from the epididymal depot. 2. Insulin increased and noradrenaline decreased [U-14C]glucose incorporation into fatty acids by brown adipocytes. Brown adipocytes differed from white adipocytes in that exogenous fatty acid (palmitate) substantially decreased fatty acid synthesis from glucose. Both noradrenaline and insulin increased lactate + pyruvate formation by brown adipocytes. Brown adipocytes converted a greater proportion of metabolized glucose into lactate + pyruvate and a smaller proportion into fatty acids than did white adipocytes. 3. In brown adipocytes, when fatty acid synthesis from [U-14C]glucose was decreased by noradrenaline or palmitate, incorporation of 3H2O into fatty acids was also decreased to an extent which would not support proposals for extensive recycling into fatty acid synthesis of acetyl-CoA derived from fatty acid oxidation. 4. In the absence of glucose, [U-14C]lactate was a poor substrate for lipogenesis in brown adipocytes, but its use was facilitated by glucose. When brown adipocytes were incubated with 1 mM-lactate + 5 mM-glucose, lactate-derived carbon generally provided at least 50% of the precursor for fatty acid synthesis. 5. Both insulin and noradrenaline increased [U-14C]glucose conversion into CO2 by brown adipocytes (incubated in the presence of lactate) and, in combination, stimulation of glucose oxidation by these two agents showed synergism. Rates of 14CO2 formation from glucose by brown adipocytes were relatively small compared with maximum rates of oxygen consumption by these cells, suggesting that glucose is unlikely to be a major substrate for thermogenesis. 6. Brown adipocytes from 6-week-old rats had considerably lower maximum rates of fatty acid synthesis, relative to cell DNA content, than white adipocytes. By contrast, rates of fatty acid synthesis from 3H2O in vivo were similar in the interscapular and epididymal fat depots. Expressed relative to activities of fatty acid synthase or ATP citrate lyase, however, brown adipocytes synthesized fatty acids as effectively as did white adipocytes. It is suggested that the cells most active in fatty acid synthesis in the brown adipose tissue are not recovered fully in the adipocyte fraction during cell isolation. Differences in rates of fatty acid synthesis between brown and white adipocytes were less apparent at 10 weeks of age.
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PMID:Lipogenesis in rat brown adipocytes. Effects of insulin and noradrenaline, contributions from glucose and lactate as precursors and comparisons with white adipocytes. 313 22

Cauda epididymal fluid (CEF) greatly stimulated the oxygen uptake of washed ejaculated ram spermatozoa; the effect was evident within 1 h and persisted over the 8 h of the experiment. The stimulus was comparable to that produced by 10 mM glucose and the effects were not additive, that is, CEF and CEF plus glucose elicited about the same oxygen uptake. This suggested that CEF suppressed the oxidation of added glucose and this was confirmed by measuring the amount of glucose oxidized in the presence and absence of CEF. Cauda epididymal fluid improved the motility of washed ram spermatozoa but it was somewhat less than that produced by glucose and usually only became evident after about 6 h or incubation. Electrophoretic analysis of cauda epididymal spermatozoa incubated in radioiodinated CEF showed that these cells absorb fluid components in the zone 82 to 56 kD. However, a molecular weight fraction less than 5 kD obtained by passing CEF through a Sephadex G-25 column, was not effective in stimulating the oxygen uptake of ram spermatozoa. The effects of CEF on the metabolism of ram spermatozoa could be mimicked by 2.5-4.0 mg/ml bovine serum albumin (BSA). Stimulation of oxygen uptake was apparent within 1 h and persisted over the 8 h of the experiment. As with CEF, stimulation of oxygen uptake by BAS was less than with 10 mM glucose but the effects were not additive. Like CEF, BSA reduced the amount of glucose oxidized. Bovine serum albumin also improved the motility of ram spermatozoa over 8 h. After passage through Sephadex G-25 to remove any low molecular weight contaminants (less than 5 kD), BSA was still effective in stimulating the oxygen uptake of spermatozoa over 4 h. Ram blood plasma and especially ram seminal plasma were also effective after passage through the Sephadex. Human serum albumin (HSA) was as effective as BSA in stimulating the oxygen uptake of ram spermatozoa but defatting decreases its effectiveness. The motility score of the spermatozoa was also adversely affected by this treatment. It is concluded that the stimulating effects of CEF and of the other fluids and proteins are due to substrates, at least some of which are present as or associated with macromolecules.
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PMID:Effect of male reproductive tract fluids and proteins on the metabolism and motility of ram spermatozoa. 343 93

A rapid decrease in male fertility in laboratory animals exposed to 1,2-dibromo-3-chloropropane (DBCP) has been suggested to be due, in part, to a postglycolytic inhibition of sperm carbohydrate metabolism. The present studies were performed to identify the specific site of DBCP-induced inhibition of intermediary metabolism. 14CO2 generation by epididymal sperm, isolated from Fischer 344 rats, was measured using radiolabeled tricarboxylic acid (TCA) cycle intermediates: acetyl CoA, citrate, alpha-ketoglutarate, and succinate. There was 0-28% inhibition of CO2 generation after addition of 0.5 mM DBCP and 81-98% inhibition with 3 mM DBCP, with all four substrates. The activities of alpha-ketoglutarate dehydrogenase, pyruvate dehydrogenase, malate dehydrogenase, and lactate dehydrogenase were not inhibited by DBCP. Since the DBCP-induced inhibition of metabolism of different substrates to CO2 was similar, and since DBCP did not inhibit enzyme activities of glycolysis or the TCA cycle, a common site of inhibition was suspected. In evaluations of mitochondrial electron transport chain activity, DBCP (3 mM) inhibited oxygen consumption resulting from metabolism of endogenous substrates plus alpha-ketoglutarate or malate by about 80%. When succinate, an FAD-dependent oxidation, was used as a substrate, oxygen consumption was not inhibited by DBCP. It is concluded that DBCP inhibits sperm carbohydrate metabolism at the NADH dehydrogenase step in the mitochondrial electron transport chain.
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PMID:A biochemical basis for 1,2-dibromo-3-chloropropane-induced male infertility: inhibition of sperm mitochondrial electron transport activity. 367 26

Sperm passing through the male tract interact with accessory sex gland fluids during ejaculation. Cellular metabolism is stimulated by this interaction for unknown reasons. These experiments involved calorimetric measurements [P.B. Inskeep and R.H. Hammerstedt (1983) J. Biochem. Biophys. Methods 7, 199-210] on ejaculated sperm (EJS) and cauda epididymal sperm (CES) from bulls to establish the contribution of individual pathways to total cellular ATP synthesis. Parallel incubations outside the calorimeter yielded samples for oxygen consumption measurements and for motility analysis, the major ATP-consuming reaction of sperm. Energy charge values were identical for incubations of EJS and CES with glucose, thereby establishing that the ratios of rates of ATP synthesis and degradation were equivalent for these cells under this incubation condition. The total rate of ATP synthesis was greater for EJS than for CES (5 vs 13 mumol ATP h-1/10(8) cells) with less than 2 mumol ATP h-1 for each cell type coming from degradation of endogenous reserves. Thus, ejaculation is associated with a large increase in catabolic rate that is satisfied by degradation of extracellular glucose. No difference in percentage of motile sperm was noted, but mean velocity was lower for CES (58 micron s-1) than for EJS (85 micron s-1). A difference in forward motility pattern was observed (wig-wag to flipping). We conclude from these data that interaction with accessory sex gland fluids alters ATP requiring activities of sperm, with one obvious alteration being their motility pattern. The increase in ATP requirement is satisfied by increased degradation of extracellular substrates, but not intracellular reserves, to provide sufficient ATP to satisfy cellular needs.
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PMID:Alterations in motility and metabolism associated with sperm interaction with accessory sex gland fluids. 402 11

The rate of TEMPONE reduction by electrons originating from ubiquinone in intact rabbit spermatozoa was observed for control, high ionic strength (HIS) medium-treated, and HIS-seminal plasma-treated (HIS-SP) samples. The presence of TEMPONE in the incubation medium had no effect on oxygen consumption, demonstrating the utility of TEMPONE as a nonperturbing probe of the ubiquinol redox state. The rate of TEMPONE reduction was significantly increased over control levels for sperm incubated in hypertonic medium and was correlated to a decrease in oxygen consumption and a relative increase in ATP in the total adenine nucleotide pool. This increase in TEMPONE reduction in HIS sperm was reversed by treatment of sperm with seminal plasma, but seminal plasma had no effect on oxygen consumption or relative amounts of ATP in the adenine nucleotide pool. These observations are consistent with state 3 respiration in control sperm and state 4 respiration in HIS- and HIS-SP-treated sperm. Arrhenius data were obtained for ejaculated and epididymal sperm subjected to a variety of treatments. Lines fitted to plots of Arrhenius data revealed that each treatment affected the activation energy and intercept relative to controls. Evidence is presented for a phase transition occurring at 13 degrees C based on changes in the rate of TEMPONE reduction by ubiquinol. It was noted that, above the phase transition, rate constants for the reaction were dependent upon both treatment and temperature, but below the transition the differential effects of treatment were no longer apparent. The present study has demonstrated that events taking place in the respiratory chain can be closely monitored by measuring oxygen uptake and TEMPONE reduction, and that these events are affected by alterations in the sperm environment.
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PMID:Alterations of oxygen uptake and the redox state of ubiquinone in rabbit sperm exposed to a variety of physiologic treatments. 408 32

1. A method is described for extracting separately mitochondrial and extramitochondrial enzymes from fat-cells prepared by collagenase digestion from rat epididymal fat-pads. The following distribution of enzymes has been observed (with the total activities of the enzymes as units/mg of fat-cell DNA at 25 degrees C given in parenthesis). Exclusively mitochondrial enzymes: glutamate dehydrogenase (1.8), NAD-isocitrate dehydrogenase (0.5), citrate synthase (5.2), pyruvate carboxylase (3.0); exclusively extramitochondrial enzymes: glucose 6-phosphate dehydrogenase (5.8), 6-phosphogluconate dehydrogenase (5.2), NADP-malate dehydrogenase (11.0), ATP-citrate lyase (5.1); enzymes present in both mitochondrial and extramitochondrial compartments: NADP-isocitrate dehydrogenase (3.7), NAD-malate dehydrogenase (330), aconitate hydratase (1.1), carnitine acetyltransferase (0.4), acetyl-CoA synthetase (1.0), aspartate aminotransferase (1.7), alanine aminotransferase (6.1). The mean DNA content of eight preparations of fat-cells was 109mug/g dry weight of cells. 2. Mitochondria showing respiratory control ratios of 3-6 with pyruvate, about 3 with succinate and P/O ratios of approaching 3 and 2 respectively have been isolated from fat-cells. From studies of rates of oxygen uptake and of swelling in iso-osmotic solutions of ammonium salts, it is concluded that fat-cell mitochondria are permeable to the monocarboxylic acids, pyruvate and acetate; that in the presence of phosphate they are permeable to malate and succinate and to a lesser extent oxaloacetate but not fumarate; and that in the presence of both malate and phosphate they are permeable to citrate, isocitrate and 2-oxoglutarate. In addition, isolated fat-cell mitochondria have been found to oxidize acetyl l-carnitine and, slowly, l-glycerol 3-phosphate. 3. It is concluded that the major means of transport of acetyl units into the cytoplasm for fatty acid synthesis is as citrate. Extensive transport as glutamate, 2-oxoglutarate and isocitrate, as acetate and as acetyl l-carnitine appears to be ruled out by the low activities of mitochondrial aconitate hydratase, mitochondrial acetyl-CoA hydrolyase and carnitine acetyltransferase respectively. Pathways whereby oxaloacetate generated in the cytoplasm during fatty acid synthesis by ATP-citrate lyase may be returned to mitochondria for further citrate synthesis are discussed. 4. It is also concluded that fat-cells contain pathways that will allow the excess of reducing power formed in the cytoplasm when adipose tissue is incubated in glucose and insulin to be transferred to mitochondria as l-glycerol 3-phosphate or malate. When adipose tissue is incubated in pyruvate alone, reducing power for fatty acid, l-glycerol 3-phosphate and lactate formation may be transferred to the cytoplasm as citrate and malate.
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PMID:The intracellular localization of enzymes in white-adipose-tissue fat-cells and permeability properties of fat-cell mitochondria. Transfer of acetyl units and reducing power between mitochondria and cytoplasm. 439 82

Chronic injections (once daily for 10-14 days) of triiodothyronine (T3) stimulated oxygen consumption by 50 and 15% in anaesthetized, control (24 degrees C), and cold-adapted (5 degrees C) rats, respectively, compared with euthyroid controls. Tissue blood flow, determined from the distribution of radioactive microspheres, was unaffected by T3 treatment in skeletal muscle, scrotum, brain, bone, skin, diaphragm, and brown adipose tissue (BAT) of rats housed at 24 degrees C, but was decreased in spleen (53% of control) and significantly increased in three white adipose tissue depots (average 267% increase) and liver (56%). Blood flow to epididymal fat and leg muscle of cold-adapted rats was increased by T3 treatment (100 and 138% increases, respectively), but other tissues were unaffected. Blood oxygen extraction and oxygen consumption in vivo by interscapular BAT was increased in hyperthyroid rats compared with euthyroid controls, but was reduced by T3 treatment in cold-adapted animals. These data show that BAT makes only a minor contribution (7%) to thyroid thermogenesis, but suggest that kidney, liver, gut, and particularly white adipose tissue may be involved.
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PMID:Tissue blood flow in control and cold-adapted hyperthyroid rats. 648 85

The characteristics of regional brown (BAT) and white adipose tissue (WAT) growth and of thermogenesis following experimental overfeeding were studied in groups of male Sprague-Dawley rats fed lab chow or cafeteria diets for 8 weeks postweaning. Regional BAT and WAT growth was determined by dissection and weighing, and thermogenesis was characterized by measurements of resting and norepinephrine (NE)-stimulated oxygen consumption, of serum thyroid hormone concentrations, and of 24-hour urinary NE excretion levels. Cafeteria feeding resulted in a 113% increase in total BAT, with the most prominent increases in the interscapular, thoracic, and perirenal regions. Retroperitoneal, epididymal, and omental WAT were significantly greater in cafeteria than in chow-fed rats. Resting oxygen consumption of cafeteria-fed rads increased by 10% and NE excretion by 64% compared to chow-fed controls, while serum T3 concentrations were nearly doubled in the cafeteria-fed rats. The thermogenic response to NE injection in cafeteria-fed rats was 102% of their resting levels, compared to a 51% increase in the chow-fed controls. The results indicate that increased BAT growth occurs in all primary BAT depots following cafeteria-feeding in rats, and that the greater BAT mass is qualitatively proportional to their greater capacity for non-shivering thermogenesis. Also, the increased NE excretion and greater serum T3 concentration are consistent with increased sympathetic and thyroidal activity and may in part explain the thermogenic response to diet in the rat.
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PMID:Characteristics of diet-induced brown adipose tissue growth and thermogenesis in rats. 707 52

Differences in the metabolism of testicular and cauda epididymal sperm have been demonstrated for several species, but the region(s) of the ram epididymis in which changes in metabolism occur is not known. In these experiments respirometric and radioisotopic methods were used to monitor metabolic activity of ram sperm isolated from six regions of the epididymis. Effects of exogenous carnitine on metabolism of the sperm also were studied. Sperm from the caput and corpus epididymidis had similar rates of glucose and oxygen consumption and of lactate and CO2 production. However, the magnitude of each parameter was severalfold higher for cauda sperm. In addition, cauda sperm produced 25 times more acetate than caput or corpus sperm. The amount of energy derived from utilization of endogenous substrates was similar for sperm from all regions of the epididymis. Exogenous D,L-carnitine (5 mM) had no effect on the metabolism of caput or corpus sperm. However, when cauda sperm were incubated with carnitine, they consumed less glucose and produced less lactate and more acetate; thus they produced the same amount of ATP from less glucose than did control aliquots incubated without exogenous carnitine. Since the rate of ATP synthesis was equivalent for both incubations, we believe this change in metabolism reflects an increased efficiency of glucose utilization. This increased efficiency may be vital for sperm motility.
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PMID:Changes in metabolism of ram sperm associated with epididymal transit or induced by exogenous carnitine. 713 17


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