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1.
Rainbow trout (Oncorhynchus mykiss) skin cell cultures were obtained by trypsinization of the tissue and grown in Leibovitz L-15 medium. Lipid class compositions, and fatty acid profiles of total lipids and individual phospholipid classes were determined at different times of culture. The metabolism of polyunsaturated fatty acids (PUFA) was investigated by incubating primary cultures after 7 and 14 days with [1-14C]18:2n-6 and [1-14C-]18:3n-3. The change in morphology between epithelial-like primary cultures and fibroblastic-like secondary subcultures was accompanied by alterations in the lipid composition. Polar lipids became predominant by 14 days in culture. The relative proportions of phosphatidylcholine (PC), the most abundant phospholipid, phosphatidylinositol and cholesterol increased significantly, while sphingomyelin decreased. Saturated fatty acids, 18:1n-9, n-6 and n-9PUFA were more abundant in total lipid in cultures at 14 days and 4 months than in cells initially isolated which contained higher percentages of longer chain monoenes and n-3PUFA. The changes in fatty acid composition with time in culture were observed in all the major phospholipid classes. Rainbow trout skin cells in culture desaturated and elongated both 18:2n-6 and 18:3n-3, with 20:4n-6 and 20:5n-3 being the most abundant products, respectively. PC presented the highest incorporation of radioactivity, especially following incubation with 18:3n-3. Lipid metabolism in general increased with the age of primary cultures, with both the amount of C18 PUFA incorporated and metabolized by desaturation/elongation significantly increased in 14 day cultures compared to 7 day cultures. Product/precursor ratios calculated for both n-6 and n-3 fatty acids showed that, while 6 desaturase activity was increased significantly with cell age, 5 desaturase activity was more affected by the fatty acid series, with 18:3n-3 being more readily transformed to 20:5n-3 than 18:2n-6 to 20:4n-6. Further desaturation of 20:5n-3 to hexaenes was low. Overall, the data suggested that the trout skin cell cultures were more similar to mammalian skin fibroblasts than mammalian epidermal/keratinocyte cultures.  相似文献   

2.
The effects of sub-lethal doses of dichlorvos and formalin, antimicrobial/parasitic agents used in aquaculture, on lipid composition and metabolism of rainbow trout skin cells in primary culture were investigated. [1-14C]Stearic (18:0), [1-14C]lin 18:2n-6) and [1-14C]linolenic (18:3n-3) acids were used as tracers to determine effects on fatty acid incorporation and metabolism. Formalin increased cell numbers and reduced the lipid content of the cells and the incorporation of radioactive fatty acids. The effects of dichlorvos were qualitatively similar but quantitatively less. Formalin induced relatively small but significant changes in lipid class composition including a decreased proportion of phosphatidycholine with increased proportions of sphatidylethanolamine and phosphatidylserine. Dichlorvos had no significant effect on lipid class compositions. The trout primary skin cells expressed substantial 9, 6 and 5 fatty acyl desaturase activities. Although, as expected, the cells were m active towards [1-14C]18:3n-3, the cells were unusually active towards [1-14C]18:2n-6. Both dichlorvos and, especially, formalin appeared to significantly inhibit 9 and 6 desaturation. Changes in the distribution of radioactivity between individual spholipid classes was also influenced by formalin and dichlorvos, and this may be related to changes in desaturase activity. This study has shown that topically active agents used in aquaculture, formalin and dichlorvos, had a range of effects on the rainbow trout skin cell cultures that may affect cell proliferation and lipid and fatty acid metabolism. Both agents significantly inhibited desaturation of fatty acids, particularly of 18:2n-6 to 20:4n-6 and, as 20:4n-6 is a major eicosanoid precursor ish and considering the importance of eicosanoids in the biochemistry of skin, it is suggested that these agents may have direct effects on fish skin that could have important consequences for fish health in general.  相似文献   

3.
Arctic charr,Salvelinus alpinus L. were fed five test diets containing 0% or 1% of different polyunsaturated fatty acids (PUFA) for 93 days. The fish were injected intraperitoneally with (1–14C)–18:2(n–6) or (1–14C)–18:3(n–3), and the bioconversion to longer chain PUFA studied. The conversion rate in neutral lipids was slow, with most label found as the fatty acid injected, while extensive modification took place prior to or during incorporation into polar lipids. Linolenic acid was preferred over linoleic acid as substrate for elongation and desaturation regardless of diet. In polar lipids, the predominant products of (1–14C)–18:2(n–6) metabolism were generally 20:3(n–6) and 20:4(n–6), while 18:4(n–3), 20:5(n–3) and 22:6(n–3) were the major products of (1–14C)–18:3(n–3) metabolism. The lack of radioactivity in 22:5(n–6) suggests that 4 desaturation is specific for (n–3) PUFA. Feeding the PUFA deficient diet reduced the 5 desaturation compared to fish maintained on PUFA supplemented diets. The 6 desaturation was only reduced in fish fed C18 PUFA and injected with (1–14C)–18:3(n–3). Longer chain C20 and C22 PUFA, particularly those of the (n–3) family, exerted some inhibition on the elongation and desaturation of injected fatty acids compared to those fed C18 PUFA. The incorporation of radiolabelled fatty acids into polar lipids of fish fed a commercial diet was very low, and the desaturation neglectible in both polar and neutral lipids, showing that Arctic charr under culture conditions do not convert short chain PUFA to longer chain metabolites.  相似文献   

4.
The incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids were studied in a cell line derived from chum salmon heart (CHH-1). Supplementing media with 25 M fatty acid considerably altered the cellular fatty acid composition but did not affect the lipid class composition or cause the appearance of cytoplasmic lipid droplets. CHH-1 cells exhibited considerable -6-desaturase activity but showed no preference between (n-3) and (n-6)PUFA substrates. CHH-1 cells also possess -5-desaturase activity which showed preference towards (n-3)PUFA, but -4-desaturase activity was totally absent. Elongation of 20-carbon PUFA was especially active in CHH-1 cells with 22-carbon PUFA being specifically incorporated into PE and PS lipid classes. The fatty acid composition of PI indicated specific incorporation of 20-carbon PUFA into this lipid class. Supplementation with 22:6(n-3) generated fatty acid compositions more closely resembling those of intact salmonid hearts. Substantial chain shortening of 22:6(n-3) to 20:5(n-3) occurred.Abbreviations BHT butylated hydroxytoluene - BSA bovine serum albumin - CL cardiolipin - FCS fetal calf serum - PA phosphatidic acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - SM sphingomyelin  相似文献   

5.
The desaturation of [1-14C]18:3n-3 to 20:5n-3 and 22:6n-3 is enhanced in an essential fatty acid deficient cell line (EPC-EFAD) in comparison with the parent cell line (EPC) from carp. In the present study, the effects of competing, unlabeled C18 polyunsaturated fatty acids (PUFA), linoleic (18:2n-6), -linolenic (18:3n-3), -linolenic (18:3n-6) and stearidonic (18:4n-3) acids, on the metabolism of [1-14C]18:3n-3 were investigated in EPC-EFAD cells in comparison with EPC cells. The incorporation of [1-14C]18:3n-3 in both cell lines was significantly reduced by competing C18 PUFA, with the rank order being 18:4n-3>18:3n-3 = 18:2n-6>18:3n-6. In the absence of competing PUFA, radioactivity from [1-14C]18:3n-3 in EPC cells was predominantly recovered in phosphatidylethanolamine followed by phosphatidylcholine. This pattern was unaffected by competing n-6PUFA, but n-3PUFA reversed this pattern as did essential fatty acid deficiency in the presence of all competing PUFA. The altered lipid class distribution was most pronounced in cells supplemented with 18:4n-3. Competing C18 PUFA significantly decreased the proportions of radioactivity recovered in 22:6n-3, pentaene and tetraene products, with the proportions of radioactivity recovered in 18:3n-3 and 20:3n-3 increased, in both cell lines. However, the inhibitory effect of competing C18 PUFA on the desaturation of [1-14C]18:3n-3 was significantly greater in EPC-EFAD cells. The magnitude of the inhibitory effects of C18 PUFA on [1-14C]18:3n-3 desaturation was dependent upon the specific fatty acid with the rank order being 18:4n-3>18:3n-3>18:2n-6, with 18:3n-6 having little inhibitory effect on the metabolism of [1-14C]18:3n-3 in EPC cells. The differential effects of the C18 PUFA on [1-14C]18:3n-3 metabolism were consistent with mass competition in combination with increased desaturation activity in EPC-EFAD cells and the known substrate fatty acid specificities of desaturase enzymes. However, the mechanism underpinning the greater efficacy with which the unlabeled C18PUFA competed with [1-14C]18:3n-3 in the desaturation pathway in EPC-EFAD cells was unclear.  相似文献   

6.
Proliferation of an essential fatty acid deficient cell line from carp (EPC-EFAD; epithelioma papillosum carp-essential fatty acid deficient) is stimulated by supplementing the cells with C20, but not C18 polyunsaturated fatty acids (PUFA). It is hypothesized that the differential ability of the PUFA to stimulate proliferation of the EPC-EFAD cells may be related to the extent of the cells' ability to desaturate and elongate C18 PUFA. In the present study, the metabolism of 14C-labeled C18 and C20 PUFA was investigated in EPC-EFAD cells in comparison with normal EPC cells. The incorporation of all the PUFA was significantly greater in EPC-EFAD cells but the rank order, 20:5n-3 > 18:3n-3 = 18:2n-6 >20:4n-6 was the same in both cell lines. The proportion of radioactivity from all labeled PUFA recovered in phosphatidylethanolamine and total polar lipids was significantly lower in EPC-EFAD cells compared to EPC cells, whereas the proportion of radioactivity recovered in all the other phospholipid classes and total neutral lipid was greater in EPC-EFAD cells. Both cell lines desaturated[1-14C]18:3n-3 and [1-14C]20:5n-3 to a greater extent than the corresponding (n-6) substrates but the desaturation of all the 14 C-labeled PUFA was significantly greater in EPC-EFAD cells compared to EPC cells. The results showed that, although essential fatty acid deficiency had several significant effects on PUFA metabolism in EPC cells, the fatty acid desaturation/elongation pathway was not impaired in EPC-EFAD cells and so they can desaturate 18:3n-3 to 20:5n-3 and 22:6n-3, and 18:2n-6 to 20:4n-6. However, 20:4n-3 and 20:3n-6, and not 20:4n-6 and 20:5n-3, were the predominant C20 PUFA produced by the elongation and desaturation of [1-14C]18:3n-3 and [1-14C]18:2n-6, respectively. Therefore, the previously reported inability of 18:3n-3 and 18:2n-6, compared to 20:5n-3 and 20:4n-6, to stimulate proliferation of the cells is apparently not due to a general deficiency in the fatty acid desaturation pathway in EPC-EFAD cells but may be related to potential differences in eicosanoid profiles in cells supplemented with C18 PUFA compared to C20 PUFA.  相似文献   

7.
Atlantic salmon (Salmo salar) were fed diets containing fish oil supplemented with 22:6n-3 (FO diet) or linseed oil supplemented with 20:5n-3 (LO diet) for 6 months. The effects of these diets, both containing about 36% n-3 fatty acids, on the esterification, desaturation and elongation of [1-14C] 18:2n-6 and [1-14C] 18:3n-3 were investigated in isolated hepatocytes. The percentages of radioactivity which was esterified from [1-14C] 18:2n-6 or [1-14C]18:3n-3 into total lipids, were approximately 20% lower in hepatocytes from fish fed the FO diet than in hepatocytes from fish fed the LO diet. The percentages of radioactivity esterified in both groups were further reduced when 0.1 mM unlabelled 22:6n-3 was added to the incubation. The percentage of desaturation and elongation products formed from [1-14C] 18:2n-6 was twice as high in hepatocytes from salmon fed the FO diet as it was in hepatocytes from fish fed the LO diet. The ratio of 18:2n-6 to 18:3n-3 was five times higher in the FO diet, and this probably promoted the conversion of 18:2n-6 to longer chain n-6 fatty acids. When 0.1mM unlabelled 22:6n-3 was added to the incubation medium, the percentages of desaturation and elongation products formed were unchanged. Thus, a high level of 22:6n-3 in the diet is apparently not inhibiting the conversion of 18:2n-6 to 20:4n-6, as long as the amount of 18:2n-6 present is substantially higher than that of 18:3n-3. No desaturation and elongation products were recovered from the phospholipids of hepatocytes incubated with [1-14C] 18:3n-3 in any of the groups. However, the `dead end' elongation product 20:3n-3 was found in the triacylglycerol fraction, and the percentage of this fatty acid increased when 22:6n-3 was added to the incubation medium.  相似文献   

8.
Tilapia (Oreochromis) nilotica were fed either a commercial diet containing 2.2% (n-3) and 0.5% (n-6) polyunsaturated fatty acids (PUFA), or a diet containing 1.0% methyl linoleate as the only PUFA. The fatty acid composition of tissue lipids generally reflected that of the diet. Fish from both dietary groups were injected intraperitoneally with 14C-labelled linoleic acid, 18:2 (n-6), or linolenic acid, 18:3 (n-3), and the distribution of radioactivity in tissue lipids examined. The conversion of both 18:2 (n-6) and 18:3 (n-3) to longer chain PUFA was lower in fish fed the commercial diet than in those fed the diet containing only 18:2 (n-6). Half of the radioactivity from both substrates recovered in liver polar lipids was present in C20 and C22 PUFA with fish maintained on the experimental diet. It is concluded that T. nilotica is capable of elongating and desaturating both 18:2 (n-6) and 18:3 (n-3), but that this conversion is suppressed by dietary longer chain PUFA. NERC Unit of Aquatic Biochemistry  相似文献   

9.
The incorporation, and the capacity for desaturation and elongation in vivo, of intraperitoneally-injected, 14C-labelled n–3 and n–6 C18 and C20 PUFAs were investigated in juvenile gilthead sea bream, Sparus aurata. The results indicate that juvenile gilthead sea bream have only limited ability to convert CH PUFAs to C20 and C22 HUFAs in vivo. The data are consistent with the results from nutritional studies on larvae, postlarvae and fingerlings that have shown that gilthead sea bream require the provision of preformed eicosapentaenoic and docosahexaenoic acids in the diet. The impairment in the desaturase/elongase pathway was quantitatively and qualitatively similar to that found in turbot, Scophthalmus maximus, being at the level of the 5-desaturase. The low activity of 5-desaturase combined with the consistent finding that arachidonic acid is selectively retained in membrane phosphatidylinositol suggests that, in addition to eicosapentaenoic and docosahexaenoic acids, gilthead sea bream may also have a requirement for preformed arachidonic acid in the diet.Abbreviations AA 5,8,11,14-eicosapenaenoic acid (arachidonic acid, 20:4n–6) - CPL diradyl (diacyl + alkenylacyl + alkylacyl) glycerophosphocholine - DHA 4,7,10,13,16,19-docosahexaenoic acid (22:6n–3) - EPA 5,8,11,14,17-eicosapentaenoic acid (20:5n–3) - EPL diradyl (diacyl, alkenylacyl + alkylacyl) glycerophosphoethanolamine - HUFA highly unsaturated fatty acids ( C20 and with 3 double bonds) - LA 9,12-octadecadienoic acid (linoleic acid, 18:2n–6) - LNA 9,12,15-octadecatrienoic acid (-linolenic acid, 18:3n–3) - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid(s)  相似文献   

10.
Accumulation of docosahexaenoic acid (DHA; 22:6n-3) in brain and eyes during development has been demonstrated in fish but it is not clear whether liver or neural tissues themselves are of greater importance in the biosynthesis of DHA from dietary 18:3n-3. In the present study, we investigated the in vivo metabolism of intraperitoneally injected [1-14C]18:3n-3 in liver, brains and eyes of young juvenile fish. Metabolism was followed over a 48h time-course in order to obtain dynamic information that could aid the elucidation of the roles of the different tissues in the biosynthesis and provision of DHA from dietary 18:3n-3. The study was performed in both a freshwater fish, rainbow trout Oncorhynchus mykiss L and a marine fish, gilthead sea bream Sparus aurata L to determine the effect that low or limiting5-desaturase activity may have in this process. As expected, the results showed that although the sea bream incorporated more 18:3n-3 into its lipids, metabolism of the incorporated fatty acid by de saturation and elongation was generally greater in the trout. In liver, the percentages of radioactivity recovered in tetraene and pentaene products were greater in trout than in sea bream although there was no difference in hexaenes. In contrast, the re covery of radioactivity in DHA was significantly greater in brain in trout compared to sea bream. In both species, the percentage of radioactivity recovered in desaturated/elongated products was much lower in liver than in brains and eyes, but that percentage increased over the 48h time-course. In trout though, the highest percentages of desaturated products in brain and eye were observed after 12 and 24h, respectively. However in sea bream the highest percentages of desaturated products in the neural tissues were observed after 24-48h. Radioactivity was recovered in 24:5n-3 and 24:6n-3, intermediates in the 4-independent ("Sprecher shunt") pathway for the synthesis of DHA, in both species, especially in the brain and eyes. Overall, although the results cannot eliminate a role for liver in the biosynthesis and provision of DHA for developing neural tissues in fish, they suggest that DHA can be synthesised in fish brain and eye in vivo.  相似文献   

11.
A marine fish oil, Marinol K (MO) and borage oil (BO) were used to formulate diets relatively rich in eicosapentaenoic acid [EPA; 20:5(n-3)] and -linolenic acid [GLA; 18:3(n-6)], respectively. The diets were fed to duplicate groups of juvenile turbot (Scophthalmus maximus) of initial weight 1.4 g for a period of 12 weeks. No differences were observed in final weights either between duplicate tanks or between dietary treatments. Mortalities in the MO-fed group were significantly greater than in the BO-fed group. In the MO-fed group, 7 out of 12 fish sampled for histological analysis showed a pronounced liver histopathology whereas only 1 of 12 fish sampled in the BO-fed group showed slight pathology. EPA levels were increased 2.2-fold and its elongation product, 22:5(n-3), was increased 1.8-fold while arachidonic acid [AA; 20:4(n-6)] was decreased by 30% in MO-fed fish compared to the initial carcass composition. GLA was increased 53-fold and its elongation product dihomo--linolenic acid [DHGLA; 20:3(n-6)] was increased 16-fold while AA was reduced by 90% in BO-fed fish compared to the initial carcass composition. The amount of triacylglycerol in liver of BO-fed fish was significantly greater than levels in MO-fed fish. The fatty acid compositions of individual phospholipids from liver showed marked differences between dietary treatments. Fish fed MO had significantly higher levels of the (n-3) polyunsaturated fatty acids (PUFA), 20:5(n-3), 22:5(n-3) and 22:6(n-3), and also significantly more 20:4(n-6) compared to BO-fed fish which had significantly higher 18:2(n-6), 18:3(n-6), 20:2(n-6) and 20:3(n-6). The composition of liver phosphatidylinositol was particularly unusual in BO-fed fish having DHGLA as the major C20 PUFA which was 2.2-fold greater than AA and 3.9-fold greater than EPA. This study demonstrates that the carcass composition of turbot can be altered, by means of dietary lipids, to contain increased levels of EPA and DHGLA which would be of potential benefit in human as well as in fish nutrition. However, caution should be exercised when using very highly unsaturated oils relatively rich in EPA which may generate histopathological lesions in the fish.Abbreviations AA arachidonic acid - ANOVA analysis of variance - BHT butylated hydroxytoluene - BO borage oil - DHA docosahexaenoic acid - DHGLA dihomo--linolenic acid - EPA eicosapentaenoic acid - GLA -linolenic acid - HPTLC high performance thin-layer chromatography - MO Marinol K - PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - TLC thin-layer chromatography  相似文献   

12.
The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in moles larva–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 g mg lipid–1 larva–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 g mg lipid–1 larva–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.  相似文献   

13.
Rainbow trout ovarian follicles were incubated in vitro with tritiated 17,20-dihydroxy-4-pregnen-3-one (17,20-P; maturation-inducing steroid). Within 18–24 h, 56–66% had been converted to tritiated 17,20-dihydroxy-4-pregnen-3-one 20-sulfate (identification confirmed by HPLC) and 27% had been taken up (absorbed) by the follicles. Addition of 125 ng of cold (non-tritiated) 17,20-P to the incubations caused a decrease in the percentage of [3H]-17,20-P which was sulfated (56% 10%) and an increase in the percentage that was taken up (27% 57%). Seven steroids were tested for their effectiveness in decreasing the sulfation and increasing the uptake of tritiated [3H]-17,20-P. The order of effectiveness was in both cases the same: 17,20-P > cortisol > 11-deoxycortisol > 17,20,21-trihydroxy-4-pregnen-3-one > 17-hydroxy-4-pregnene-3,20-dione > 17-estradiol > testosterone. This indicated that the processes of sulfation and uptake of [3H]-17,20-P were related to each other and led to the hypothesis that, when cold 17,20-P is added to the medium, it reduces the proportion of [3H]-17,20-P which is sulfated and thus allows more free [3H]-17,20-P to enter the ovarian follicles. This hypothesis was supported by the finding that each ovarian follicle had the capacity in vitro to sulfate only ca. 2 ng of [3H]-17,20-P per 18h but a capacity to take up > 500 ng per 18h.Gonadotropin I, Gonadotropin II, forskolin and phorbol-12-myristate-13-acetate (which all have an affect on steroid biosynthesis) did not affect the amount of 17,20-P which was sulfated. Sulfating activity was localized in the thecal cell layer of the follicle. The yolk fraction was shown to be responsible for absorbing the [3H]-17,20-P.  相似文献   

14.
Cells from a relatively stenohaline marine species, turbot (Scophthalmus maximus) (TF) and an anadromous species, Atlantic salmon (AS) were cultured in media supplemented with NaCl to produce OPs varying from 300 to 500 mOsm kg–1 and the direct effects of OP (salinity) on the fatty acid compositions of the main glycerophospholipid classes were determined. The most dramatic effects of salinity on total lipid fatty acids were observed in polyunsaturated fatty acids (PUFA) in TF cells. There was a graded decrease in the percentage of 18:2n-9, and consequently total n-9 PUFA, and concomitantly increased percentages of both total n-3 and n-6 PUFA with increasing salinity. The increased n-3 and n-6 PUFA was due to significantly increased percentages of the major fatty acids in each of these groups, namely 22:6n-3 and 20:4n-6, respectively. The reciprocal changes in n-9 PUFA and n-3/n-6 PUFA in TF cell total lipid resulted in the percentage of total PUFA not being significantly affected by changes in salinity. The graded decrease in 18:2n-9 with increasing salinity in TF cells was observed in all the major glycerophospholipids but especially PE, PI and PS. Increasing salinity resulted in graded increases in the percentages of 22:6n-3 in PE and PS in TF cells. The quantitatively greatest increase in the percentage of n-6 PUFA in TF cells occurred with 20:4n-6 in PC, PE and PL. There were less significant changes in the fatty acid compositions of glycerophospholipids in AS cells. However, the proportion of total n-3 + n-6 PUFA in PE varied reciprocally with the proportion of dimethylacetals in response to salinity. Similar reciprocal changes between fatty acids in response to salinity were also evident in the quantitatively more minor glycerophospholipids PS and Pl. In PS, the percentage of 22:6n-3 was significantly lower at 400 mOsm kg–1 whereas the proportion of total monoenes was significantly higher at that salinity. A similar inverse relationship between total monoenes and 20:4n-6 (and, to a lesser extent total saturates) in response to salinity was noted in PI. The results show that environmental salinity, without whole-body physiological stimuli, has direct effects on the fatty acid composition of major glycerophospholipid classes in fish cells and that these effects differ in cells from different fish speciesAbbreviations ANOVA analysis of variance - BHT butylated hydroxytoluene - BSA bovine serum albumin - DMA dimethylacetals - EMEM Eagle's minimal essential medium - FCS fetal calf serum - GC gas chromatography - HBSS Hank's balanced salt solution (without Ca2+ and Mg2+) - OP osmotic pressure - PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - TLC thin-layer chromatography  相似文献   

15.
Arachidonic acid (AA; 20:4n-6) is the precursor of a range of highly biologically active derivatives, collectively termed eicosanoids, including prostaglandins, thromboxanes, leukotrienes and lipoxins, that act as autocrine hormones regulating many physiological processes including haemostasis, reproduction, immune and inflammatory responses. Eicosapentaenoic (EPA; 20:5n-3) and dihomo-γ-linolenic (20:3n-6) acids modulate eicosanoid metabolism by both inhibiting the conversion of AA to eicosanoids whilst simultaneously being converted to eicosanoids with different, often attenuated, properties compared to their AA homologues. Eicosatetraenoic acid (20:4n-3) is a naturally occurring C20 polyunsaturated fatty acid (PUFA), present in fish oil at levels of around 1–2%, that has been suggested to be the active metabolite responsible for the anti-inflammatory effects of plant oils containing stearidonic acid (18:4n-3). However, the biochemical properties of 20:4n-3 in terms of cellular biology have rarely been investigated, partly due to difficulties in obtaining the fatty acid in high purity. In this paper, we describe methods for the medium scale laboratory preparation of high purity 20:4n-3, and investigate its metabolism in fish cell culture systems which normally contain significant amounts of n-3 PUFA. Thus the incorporation and metabolism of 18:4n-3 and 20:4n-3, and their distribution in phospholipid classes was studied in an established cell line from Atlantic salmon (Salmo salar) (AS), and the effects of 20:4n-3 on eicosanoid production studied in freshly isolated macrophages from rainbow trout (Oncorhynchus mykiss). Both 18:4n-3 and 20:4n-3 were preferentially esterified into phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine in contrast with the accumulation of AA in phosphatidylinositol. Incorporated 18:4n-3 was readily converted to 20:4n-3, and both fatty acids were further desaturated and elongated to EPA and 22:5n-3 but not 22:6n-3. Supplementation with 20:4n-3 decreased the conversion of AA into prostaglandins, as demonstrated by the decreased levels of PGF2α produced in trout macrophages supplemented with 20:4n-3 and AA compared to cells supplemented with AA alone. In addition, 20:4n-3 was converted into eicosanoids in fish cells as indicated by the presence of Δ17,18 12-HETE, Δ17,18 PGE1 and Δ17,18 PGF1α in extracts from rainbow trout macrophages incubated with 20:4n-3.  相似文献   

16.
Spawners of the great scallop, Pecten moximus, were conditioned on three microalgal diets: T-Isochrysis, a mixture of four species (PTSC) and Chaetoceros calcitrans.The polyunsaturated fatty acid (PUFA) composition of neutral and polar lipids of eggs was related to the fatty acid composition of the diet. However, the 20 and 22 carbon PUFAs were maintained at significant levels independent of those of the diet when these fatty acids were present in the diet at low levels. This effect was more pronounced in the polar than in the neutral lipids. A preferential incorporation of 22:6(-3) and 20:4(-6) was demonstrated in the polar lipids. This emphasizes their role in ovogenesis and embryogenesis.The 22:6(-3) requirement of P. maximus was better satisfied by T-Isochrysis, which favoured the highest incorporation of this essential fatty acid in the eggs. The good success in reproduction obtained with this monospecies diet led us to modify multispecies diets by increasing the level of 22:6(-3).  相似文献   

17.
In each of two separate experiments, eggs from a single female goldfish were fertilized, incubated at 22°C and sampled regularly up to day 6 when the larvae could be expected to commence feeding. Hatching normally occurred on Day 4. Lipids were extracted from the eggs and larvae and the neutral lipid and neutral phospholipids were isolated on aminopropyl columns. Fatty acid analysis of the eggs revealed the typical situation in fish where the phospholipids were rich in polyunsaturated fatty acids (PUFA) and the neutral lipids were rich in monounsaturated fatty acids (MUFA). Assay of lipid masses revealed that little depletion of lipid occurred until after hatch and that the neutral phospholipids were the principal fraction consumed. Although the neutral lipid mass did not change substantially during development, its fatty acid profile did. The proportions of several PUFA in the neutral lipids, especially 226(n–3), 205(n–3) and 204(n–6), increased substantially during development while proportions of MUFA and 182(n–6) declined. This appears to be a mechanism by which the larva can retain essential fatty acid released on hydrolysis of phospholipid while deriving the benefits of catabolism of phospholipid as fuel, namely the provision of phosphate and choline for intermediary metabolism and for the synthesis of macromolecules and neurotransmitter.Abbreviations AA arachidonic acid (204(n–6)) - DHA docosahexaenoic acid (226(n–3)) - EPA eicosapentaenoic acid (205(n–3)) - MUFA monounsaturated fatty acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PUFA polyunsaturated fatty acid - SFA saturated fatty acid  相似文献   

18.
The fatty acid compositions of wild female northern pike (Esox lucius L.) and their principle prey species were compared to assess the extent to which pike modify the relative abundance of dietary fatty acids during assimilation and to indicate the optimum dietary content of essential fatty acids (EFAs) for pike. Only minor differences existed between the estimated whole body fatty acid composition of pike and diet fatty acid composition as calculated from the contribution of each prey species to the pike's diet. Saturated fatty acids comprised a slightly higher percentage of diet lipids (25% wt) than of pike lipids (21% wt) whereas monounsaturated fatty acids were less abundant in diet lipids (26% wt) than in pike (29% wt). Percentages of total polyunsaturated fatty acids (PUFAs), n - 3 fatty acids, and n - 6 fatty acids were approximately 43, 30, and 13% wt respectively and differed by less than 1% wt between pike and diet lipids. Among individual PUFAs, the largest differences occurred in 20:5 (n-3) and 22:6(n-3) which comprised, on average, 9.6 and 14.7% wt respectively of diet lipids and 5.9 and 18.3% wt respectively of pike lipids. The close similarity in fatty acid composition between pike and their diet suggests that pike may have limited abilities to elongate and desaturate 18 carbon PUFAs and may require specific long chain PUFAs in the diet. The n-3 PUFA content of the pike's natural diet may exceed the minimum EFA requirements of better studied species such as rainbow trout and turbot.  相似文献   

19.
The fatty acid compositions of brain phosphoglycerides from a freshwater fish, the rainbow trout (Salmo gairdneri), and a marine fish, the cod (Gadus morhua), were determined and compared with those from a terrestrial mammal, the rat. Fish brain lipids were characterized by a higher degree of unsaturation encompassing increased percentages of (n–3)PUFA (226 and 205) and lower percentages of (n–6)PUFA (204 and 224). However the distribution of fatty acids and specific PUFA between different phosphoglycerides was essentially similar in rat and fish brain tissue. PE and PS contained the highest percentages of 226(n–3), PI was characterized by higher 180 and 204(n–6)/205(n–3), and PC had higher 160 and the lowest percentage of PUFA in all species. A generally similar pattern was found in the fish retinal phosphoglycerides except that PC was also rich in 226(n–3). Overall trout brain phosphoglycerides were slightly more unsaturated than the cod lipids but with lower (n–3)/(n–6) ratios whereas cod retinal lipids were more unsaturated than the trout retinal lipids.  相似文献   

20.
Despite the potential of vegetable oils as aquafeed ingredients, a major drawback associated with their utilization is the inferior level of beneficial n-3 long-chain polyunsaturated fatty acids (LC-PUFA). Echium oil (EO), which is rich in stearidonic acid (SDA, 18:4n-3), could potentially improve the deposition of n-3 LC-PUFA as the biosynthesis of LC-PUFA is enhanced through bypassing the rate-limiting ?6 desaturation step. We report for the first time an attempt to investigate whether the presence of a desaturase (Fads2) capable of ?4 desaturation activities and an elongase (Elovl5) will leverage the provision of dietary SDA to produce a higher rate of LC-PUFA bioconversion. Experimental diets were designed containing fish oil (FO), EO or linseed oil (LO) (100FO, 100EO, 100LO), and diets which comprised equal mixtures of the designated oils (50EOFO and 50EOLO) were evaluated in a 12-week feeding trial involving striped snakeheads (Channa striata). There was no significant difference in growth and feed conversion efficiency. The hepatic fatty acid composition and higher expression of fads2 and elovl5 genes in fish fed EO-based diets indicate the utilization of dietary SDA for LC-PUFA biosynthesis. Collectively, this resulted in a higher deposition of muscle eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) compared to LO-based diets. Dietary EO improved the ratio of n-3 LC-PUFA to n-6 LC-PUFA in fish muscle, which is desirable for human populations with excessive consumption of n-6 PUFA. This study validates the contribution of SDA in improving the content of n-3 LC-PUFA and the ratio of EPA to arachidonic acid (ARA, 20:4n-6) in a freshwater carnivorous species.  相似文献   

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