Properties of cod metallothionein,its presence in different tissues and effects of Cd and Zn treatment

One isoform of the low-molecular-weight metal-binding protein metallothionein (MT) has been isolated from the liver of Atlantic cod by size-exclusion and ion-exchange chromatography. Cod MT contained 33% cysteine, no aromatic amino acids or arginine. As is the case for other piscine MTs, the N-terminus of cod MT lacked the asparagine in position 4 which is present in mammalian MTs. In addition, cod MT differed from all other vertebrate MTs described in that the N-terminal methionine was not acetylated. Antibodies were raised in rabbits against hepatic MT from cod by repeated injections of native protein mixed with adjuvant. Anti-cod MT antisera cross reacted with similarly-sized proteins in liver, brain, anterior kidney, posterior kidney, spleen, intestine, gills and ovaries. The putative MT in cod brain migrated differently to that of the other tissues in native gel electrophoresis. Intraperitoneally injected Cd (1 mg/kg) was nearly entirely associated with the MT-peak in hepatic and renal cytosols, whereas a single injection of Zn (10 mg/kg) resulted in increases in all cytosolic Zn pools of the liver and no apparent change in cytosolic Zn, Cu, Ni or Cd in kidney.     

饲粮中添加葵花籽油和茶油对泌乳水牛生产性能及乳脂脂肪酸组成的影响

本文旨在研究饲粮中添加葵花籽油和茶油对泌乳水牛生产性能及乳脂脂肪酸组成的影响。选取32头健康泌乳水牛(平均年龄为6.0岁,平均体重为620 kg),按产奶量(9 kg/d以上)、泌乳期(处于泌乳中期)相近原则随机分为4组,进行为期6周的饲养试验,其中预试期2周,正试期4周。基础饲料由精料和粗料组成。对照组饲喂基础饲粮;在试验组的精料中分别添加4%的葵花籽油(葵花籽油组)、4%的茶油(茶油组)、2%的葵花籽油+2%的茶油(混合组)。结果表明:与对照组相比,各试验组的产奶量均显著下降(P<0.05),乳脂率均显著提高(P<0.05),乳脂产量、乳蛋白率和乳糖率则无显著变化(P>0.05)。与对照组相比,葵花籽油组、茶油组和混合组的乳总固形物含量分别提高了5.22%、7.95%和6.17%,其中茶油组和混合组与对照组差异显著(P<0.05)。与对照组相比,葵花籽油组乳脂中cis9,trans11-C18∶2[共轭亚油酸(CLA)]的含量提高了44.54%(P<0.05),混合组乳脂中trans11-C18∶1的含量提高了22.68%(P<0.05)。葵花籽油组、混合组乳脂中C18∶2的含量均显著高于对照组(P<0.05);葵花籽油组、茶油组和混合组的多不饱和脂肪酸(PUFA)/饱和脂肪酸(SFA)均比对照组提高,且葵花籽油组和混合组显著高于对照组(P<0.05),其中混合组的PUFA/SFA最高。各组水牛的血液总胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白、葡萄糖、总蛋白、尿素氮含量及谷丙转氨酶、谷草转氨酶活性差异不显著(P>0.05)。由此得出,泌乳水牛饲粮中添加精料饲喂量4%的葵花籽油、4%的茶油或2%的葵花籽油+2%的茶油合均可提高乳脂率和乳脂中CLA的含量,但会导致产奶量降低;优化乳脂脂肪酸组成上,以葵花籽油和茶油的混合添加效果较好。     

Effects of synthetic kainic acid on microfilament expression and gap junction in astrocytes

AIM：To study the effects of synthetic kainic acid (SKA) and 1-heptanol (1-Hep), a gap junction blocker, on the cytoskeletal filament expression in the astrocytes. METHODS：The neonatal rat brain was obtained from the Wistar rats (1 day old) and primary purified astrocytes were obtained by differential attachment for removing filamentoblasts and orbital shaker for removing the oligodendrocytes. The effects of SKA and other interventions on the morphologic changes and expression levels of skeleton protein filamentous actin (F-actin) were observed in the astrocytes after 24 h of the exposures by laser scanning confocal microscopy. The effect of 1-Hep on the expression of F-actin was also explored. RESULTS：Compared with control group, the fluorescence intensities in KCl group and KCl+SKA group were increased, and highly increased in KCl+SKA group. The F-actin filaments in the above 2 groups were more intensive, thickened and concentrated than those in control group, and more obvious in KCl+SKA group. l-Hep significantly decreased the expression of F-actin in KCl group and KCl+SKA group as compared with control group, and parts of the filamentous fracture were seen in the astrocytes in all 1-Hep-treated groups, in which some of the filamentous lines were crosscut. CONCLUSION：Increase in the expression of F-actin in the astrocytes affects the structure and function of the intercellular gap junctions, which may be involved in the mechanism of SKA-induced epilepsy.     

Effect of retinoic acid on gene expression profiles of bovine intramuscular preadipocytes during adipogenesis

To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation‐ and proliferation‐phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all‐trans retinoic acid (ATRA). Quantitative real‐time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3‐L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty‐eight hours after transfection, the messenger RNA expression level of the adipose differentiation‐related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)‐transfected cells was 3.5% of that in negative control‐transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA‐transfected cells was 21.8% lower than that in negative control‐transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.     

Effect of diacylglycerol acyltransferase 2overexpression in 3T3-L1 is associated to an increase in mono-unsaturated fatty acid accumulation

Background：Fatty acid（FA） composition is the most important parameter affecting the flavor and nutritional value of the meat.The final and the only committed step in the biosynthesis of triglycerides is catalyzed by diacylglycerol acyltransferase 2（DGAT2）.The role of DGAT2 in lipid accumulation has been demonstrated in adipocytes,However,little is known about the effect of DGAT2 on the FA composition of these cells.Methods：To investigate the role of DGAT2 in regulating lipid accumulation,FA composition and the expression of adipogenic genes,we cloned the open reading frame of the porcine DGAT2 gene and established 3T3-L1 cells that overexpressed DGAT2.Cells were then cultured in differentiation medium（DM） without FA,with a mixture of FAs（FA-DM）,or containing a ~（13）C stable isotope-labeled FA mixture（IFA-DM）.The FA composition of adipocytes was analyzed by gas chromatography-mass spectrometry and gas chromatography-isotope ratio mass spectrometry.Quantitative PCR and western blotting were employed to detect expression of adipogenic genes in 3T3-L1 adipocytes cultured with FA-DM for 12 d.Results：The triacylglyceride（TAG） content was significantly higher in 3T3-L1 adipocytes overexpressing DGAT2 than in control cells.When cultured in DM or FA-DM for 12 d,cells overexpressing DGAT2 showed a higher proportion of unsaturated FAs（C16：1 and C18：1）.However,when cells overexpressing DGAT2 were cultured with FA-DM for30 min,the FA composition was almost identical to that of controls.Further,the proportion of stable isotope-labeled FAs were similar in 3T3-L1 adipocytes overexpressing DGAT2 and control cells cultured in IFA-DM for 12 d.These results collectively indicate that the higher proportion of mono-unsaturated FAs,C16：1 and C18：1,may originate from de novo FA synthesis but not from the uptake of specific FAs from the medium.This hypothesis is further supported by evidence that both mRNA and protein expression of genes involved in FA synthesis（ACACA,FASN,SCD1,and A-FABP?     

Dietary essentiality of ascorbic acid in rohu larvae: Quantification with ascorbic acid enriched zooplankton

Carp larvae, like any other fish larvae dependon natural food during first few days of theirlife. In nursery conditions, high mortality andslow larval growth are of common occurrence;sub-optimal nutrition might be a possiblereason for such consequences. To improve thesituation the effect of feeding ascorbicacid-enriched live food on survival, growth,tissue biochemical composition includingascorbate level was evaluated in first feeding(3 days old) larvae (av. wt. 2.2 mg) of therohu carp, Labeo rohita (Ham.) for aperiod of 15 days (temp. 28.6 ± 1 °C)under natural photoperiod. The larvae (stockingdensity 10 l^–1) were offered enriched andnon-enriched zooplankton ad libitumfollowing a rigid schedule with four feedingregimes, each having 3 replicates. In treatmentT₁, non-enriched zooplankton (Moina,Daphnia, Cyclops, Diaptomus) and in T₂,T₃, T₄ ascorbic acid enriched (12 henrichment) zooplankton [@10%, 20% and 30%ascorbyl palmitate (AP) inclusion in diet ofzooplankton] were offered. Highest survival(90%) and growth (9563% live weight gain)could be seen in T₃ group and the lowestin T₁ (62% survival and 805% live weightgain), thus confirming the dietary essentialityof ascorbic acid for rohu larvae. Therequirement has been shown to be 1409 µg/gdry diet. Whole body tissue analyses for crudeprotein, total lipid and RNA: DNA ratiofollowed the same trend as that of growthresponse and percent survival. Significantpositive correlation (r = 0.949 and 0.861) couldbe found with muscle RNA/DNA ratio and muscleRNA content with specific growth rate indifferent treatments. Significant differencewas found in tissue ascorbate levels betweenenriched plankton fed groups, being highest in T₃. Such live foodmediated vitamin transfer might be an effectivemeans to provide higher plane of nutrition forhigh survival and rapid growth for rohu larva.     

Artemia prey size and mode of presentation: Effects on the survival and growth of phyllosoma larvae of southern rock lobster (Jasus edwardsii)

Performance of phyllosoma of thesouthern rock lobster (Jasus edwardsii)was examined after feeding Artemia-baseddiets. Survival and growth of newly-hatchedlarvae cultured to Stage III were lower(p < 0.05) when fed 0.8 mm Artemia than1.5 mm or 2.5 mm Artemia alone or 1.5 mmArtemia in combination with pieces ofmussel (Mytilus edulis planulatus) gonad.This could not be attributed to deficiencies inthe composition of fatty acids but appeared tobe due to the inability of larvae to capturesufficient appropriate-sized, enrichedArtemia for their nutritional requirements.There was an indication that survival andgrowth were higher between Stages III and Vwhen fed 2.5 mm Artemia than 1.5 mmArtemia alone or in combination with musselpieces. However, Stage VI larvae grew to asimilar size at Stage VIII when fed 1.5 mm or2.5 mm Artemia. Unexpectedly, larvae fedthe combination of 1.5 mm Artemia plusmussel supplement had lower survival than foundpreviously, and generally lower than when fed 1.5 mm Artemia alone. This was despitean apparent nutritional profile (lipid contentand fatty acid composition) of mussel more akinto that of newly-hatched phyllosoma thanenriched Artemia. On the other hand,survival and growth to Stage VIII were higherwhen larvae were fed alginate pelletscontaining Artemia than when fed 1.5 mmor 2.5 mm Artemia alone.