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Biomedical subjects

J G Coniglio

Publications and source records attributed to J G Coniglio.

At least 37 records · Page 2Linked to original sources

Fatty acid synthesis in testes of fat-deficient and fat-supplemented rats.

Fatty acid synthesis was studied in testes of rats fed a fat-free or fat-supplemented diet. Testes of fat-deficient rats incorporated nearly twice as much intratesticularly injected [1-14C]acetate into total fatty acids (primarily into palmitic acid) as did supplemented rats. To determine the mechanism for the increased synthesis, the activities of the following enzymes were determined in the cytoplasmic fraction of testicular homogenates: fatty acid synthetase, acetyl CoA carboxylase [EC 6.4.1.2], citrate-cleavage [EC 4.1.3.8], malic [EC 1.1.1.38], and the glucose-l-phosphate dehydrogenase [EC 1.1.1.49]: 6-phosphogluconate dehydrogenase pair [EC 1.1.1.44]. Although the activity of fatty acid synthetase did increase in livers from fat-deficient rats, no change was observed in corresponding testes. No difference between the two groups could be demonstrated in testicular activity of citrate-cleavage enzyme, malic enzyme, or the glucose-6-phosphate dehydrogenase: 6-phosphogluconate dehydrogenase pair. However, the activity of cytoplasmic acetyl CoA carboxylase in testes of rats fed the fat-deficient diet was 1.4 times higher than the activity in testes of rats fed the supplemented diet. Fat deficiency did not affect the specific activity of the testicular microsomal elongation system, assayed by incubation with 14C-malonyl CoA. The concentration of unesterified fatty acids was lower in testes of the fat-deficient compared to supplemented rats, indicating that decreased inhibition of acetyl CoA carboxylase in the fat-deficient rats testes might have been responsible for the observed increased de novo synthesis of palmitic acid.

ATP Citrate (pro-S)-Lyase↗

The metabolism of (1-14C) stearic acid in rat testicular tissue.

(1) The metabolism of stearic acid was studied in vivo following intratesticular injection of [1-14C] stearate. Soon after injection 14C activity was found mainly in the free fatty acid pool. This was followed at later time periods by transfer of label primarily to the phosphatide pool. During each time period significant amounts of label were recovered at 14CO2. (2) Analysis of 14C-labeled fatty acids from the injected testes demonstrated an initial rapid rate of oxidation and desaturation of [1-14C] stearate followed by a slower steady state rate. It was concluded that the initial rate was due to the rapid turnover of the highly labeled free fatty acid pool followed by a much slower rate as [14C] stearate was esterified to the more metabolically stable phospholipids. Elongation of the labeled stearic or its desaturated derivative was not observed. (3) The rate of desaturation in vitro of stearic acid was measured in microsomal preparations from rat testes and found to be 12.0 +/- 0.5 pmol/min/mg compared to the estimated in vivo value of 22 pmol/min/mg and the value of 390 pmol/min/mg for hepatic microsomal desaturase.

Animals↗

Effect of eicosa-5, 8, 11, 14-tetraynoic acid on fatty acid composition of selected organs in the rat.

Eicosa-5, 8, 11, 14-tetraynoic acid or archidonic acid or no supplement (controls) was given orally to rats maintained on a fat free diet and the fatty acid composition of total lipids of several organs determined. No changes were noted in the total fatty acid concentrations of the organs examined in the various groups. A decrease in the amount of arachidonic acid, 22:4omega6 and 22:5omega 6 (as percent of total fatty acids), and an increase in the amount of 20:3omega6 and linoleate were observed in total lipids of several organs. In the group receiving the arachidonate supplement, there was less linoleate and 20:3omega 6 and more archidonate than in the controls. Both eicosa-5, 8, 11, 14-tetraynoic acid and arachidonate supplements resulted in a decrease in 20:3omega9 in most organs studied. Generally, the most marked changes were seen in liver but, of the other organs examined (heart, kidney, testis, brain, and adrenals), only the adrenals failed to show any significant differences between the controls and each of the two supplemented groups. Although the experimental conditions preclude conclusive interpretation of the changes observed, it is suggested that eicosa-5, 8, 11, 14-tetraynoic acid was effective in inhibiting the conversion of linoleate to archidonate and the conversion of arachidonate to 22:4 and 22:5.

5,8,11,14-Eicosatetraynoic Acid↗

Lipid and fatty acid composition of testes of quaking mice.

Testes of quaking mice (sterile mutants) and of controls were analyzed for major lipid classes and fatty acid composition. Of the main lipid classes, only cholesterol esters differed significantly in concentration between the two groups (1.01 for quakers vs 0.69 mg/g wet wt of tissue for controls). The concentration of triglycerides was 4.5-5.0 that of total phosphatides 18-19 and that of free cholesterol 1.9-2.0 mg/g for mutants and controls. The concentrations of phosphatidyl ethanolmanine and of sphingomyelin were both lower in quaking than in normal mice, but only the change in the former was statistically significant. Phosphatidyl choline was the major phosphatide (43-45% of total phosphatides) followed by phosphatidyl ethanolamine (24-26%) and sphingomyelin, phosphatidyl serine, and phosphatidyl inositol (all ca. 7% of total phosphatides). Minor differences between the mutants and controls were observed in concentrations of fatty acids of major lipid classes. The mutants, sterile because of faulty spermatid differentiation, had normal quantities of 22:6 w3 and 22:5 w6. These data are consistent with the hypothesis that the 22-carbon polyenes are associated with the formation of spermatids, rather than with their final differentiation into spermatozoa.

Animals↗

Studies in vitro of lipogenesis in rat testicular tissue.

Testicular tissue was shown to contain the full complement of enzymes required for de novo synthesis of fatty acids. The enzymes capable of snythesizing palmitic acid from citrate, acetate, or acetyl CoA were found to be present in the soluble (cytoplasmic) fraction. These included fatty acid synthetase, acetyl CoA carboxylase, citrate-cleavage enzyme, malic enzyme, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase. Optimal conditions for assaying activities of fatty acid synthetase and acetyl CoA carboxylase in the soluble fraction from rat testes were established, and the activities of these two enzymes were determined to be 0.54 +/- 0.1 and 0.030 +/- 0.002 (nmoles of substrate incorporated into fatty acid per min per mg of soluble fraction protein), respectively. The activities of citrate-cleavage enzyme, malic enzyme, and the glucose-6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase pair were also measured. The activities were 6.0 +/- 0.7, 34.9 +/- 4.2, and 29.9 +/- 9.3 nmoles/min/mg, respectively.

ATP Citrate (pro-S)-Lyase↗

Fluorescent products and polyunsaturated fatty acids of human testes.

Lipid soluble fluorescent pigments from human testis were fractionated by silicic acid column chromatography and silica gel thin layer chromatography. Fluorescence analyses revealed a family of at least 3 compounds with similar fluorescence properties, including excitation and emission maxima, reversible fluorescence quenching by alkaline pH, and fluorescence quenching by heavy metal chelation. These fluorescence characteristics strongly indicated the presence of the conjugated Schiff base fluorophore-N=C-C=C-N-. The chromatographic separations employed enabled a more definitive fluorescence characterization of the lipid soluble pigments known to accumulate late in tissues with age and as a result of lipid peroxidation. Total lipids and fatty acid composition of the total lipids were determined. Polyenoic acids constituted about 40% of the total fatty acids. Histological examination of the tissues revealed some degeneration and edema, but significant spermatogenesis and normal complement of Leydig cells.

Age Factors↗