Effects of sex on meat quality traits, amino acid and fatty acid compositions, and plasma metabolome profiles in White King squabs.

The objective of this study was to investigate the effects of sex on meat quality and the composition of amino and fatty acids in the breast muscles of White King pigeon squabs. Untargeted metabolomics was also conducted to distinguish the metabolic composition of plasma in different sexes. Compared with male squabs, female squabs had greater intramuscular fat (IMF) deposition and lower myofiber diameter and hydroxyproline content, leading to a lower shear force. Female squabs also had higher monounsaturated fatty acid and lower n-6 and n-3 polyunsaturated fatty acid proportions in the breast muscle, and had greater lipogenesis capacity via upregulation of PPARγ, FAS and LPL gene expression. Moreover, female squabs had lower inosine 5'-monophosphate, essential, free and sweet-tasting amino acid contents. Furthermore, Spearman's correlations between the differential plasma metabolites and key meat parameters were assessed, and putrescine, N-acetylglutamic acid, phophatidylcholine (18:0/P-16:0) and trimethylamine N-oxide were found to contribute to meat quality. In summary, the breast meat of male squabs may have better nutritional value than that of females, but it may inferior in terms of sensory properties, which can be attributed to the lower IMF content and higher shear force value. Our findings enhance our understanding of sex variation in squab meat quality, providing a basis for future research on pigeon breeding.

Research Note: Effect of different photoperiodic programs from rearing period on the reproductive performance and hormone secretion of White King pigeons.

The photoperiod is an important factor during rearing and laying period that affects age and body weight at sexual maturation and reproductive performance in poultry; however relevant research on this factor in pigeons is still lacking. Thus, this study investigated the effects of different photoperiodic programs on the reproductive performance and hormonal profile in White King pigeons. From 101 d of age, the pigeons in the control group were exposed to a natural photoperiod until 160 d, and then to a photoperiod of 16 h (16 light [L]: 8 dark [D]) and lasted for 200 d. Pigeons in the 3 experimental groups were exposed to a short photoperiod of 8L: 16D until 160 d, and then to 14L: 10D, 16L: 8D, and 18L: 6D, respectively. The results showed that light-restriction (8L: 16D) during the rearing period and then 14L: 10D or 16L: 8D photostimulation delayed the age at first egg laying in pigeons. However, 16L: 8D after an 8L: 16D photoperiod during the breeding period ensured maximum photosensitivity, and significantly improved the reproductive performance (egg production and fertility rates) in pigeons. Moreover, the highest reproductive performance in group under16L: 8D after 8L: 16D photoperiodic program was accompanied by improved follicle-stimulating hormone and estradiol levels and reduced prolactin hormone levels. The results indicated that photoperiodic programs from rearing to laying period are closely related to the reproductive performance of White King pigeons. The results provide information that 8L: 16D during rearing period and 16L: 8D during laying period can be used to enhance reproductive performance in the pigeon industry.

Effect of Heat Stress on Egg Production, Steroid Hormone Synthesis, and Related Gene Expression in Chicken Preovulatory Follicular Granulosa Cells.

This study was conducted to elucidate the molecular mechanisms underlying heat stress (HS)-induced abnormal egg-laying in laying hens. Hy-Line brown laying hens were exposed to HS at 32 °C or maintained at 22 °C (control) for 14 days. In addition, granulosa cells (GCs) from preovulatory follicles were subjected to normal (37 °C) or high (41 °C or 43 °C) temperatures in vitro. Proliferation, apoptosis, and steroidogenesis were investigated, and the expression of estrogen and progesterone synthesis-related genes was detected. The results confirmed that laying hens reared under HS had impaired laying performance. HS inhibited proliferation, increased apoptosis, and altered the GC ultrastructure. HS also elevated progesterone secretion by increasing the expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), and 3b-hydroxysteroid dehydrogenase (3ß-HSD). In addition, HS inhibited estrogen synthesis in GCs by decreasing the expression of the follicle-stimulating hormone receptor (FSHR) and cytochrome P450 family 19 subfamily A member 1 (CYP19A1). The upregulation of heat shock 70 kDa protein (HSP70) under HS was also observed. Collectively, laying hens exposed to high temperatures experienced damage to follicular GCs and steroidogenesis dysfunction, which reduced their laying performance. This study provides a molecular mechanism for the abnormal laying performance of hens subjected to HS.

Monochromatic Green Light Stimulation during Incubation Alters Hepatic Glucose Metabolism That Improves Embryonic Development in Yangzhou Goose Eggs.

The influence of monochromatic green light stimulation on hatching performance and embryo development has been studied in chickens, but not geese. The liver has crucial functions in the regulation of energy metabolism during embryogenesis, but its involvement in green light transduction is still unidentified. We aimed to determine the influence of monochromatic green light on Yangzhou goose hatching performance and embryo development. We also investigated the metabolomics and transcriptomic responses of the embryonic liver to green light to determine the underlying molecular mechanisms. Eggs were incubated under either 12 h of monochromatic green light/dark (12 L:12D) cycles or 24 h of darkness (0G:24D). Green light promoted embryonic development and hatching performance, also affected the expression of myogenic regulatory factors associated with muscle development. It also shortened hatching time and elevated plasma levels of growth hormone and insulin-like growth factor-1. Metabolomics and transcriptomic results revealed differentially expressed genes and metabolites with enhanced gluconeogenesis/glycolysis and increased plasma glucose and pyruvate levels under green light. Hence, the growth-promoting effect possibly through regulating energy metabolism in the liver and myogenic regulatory factors in muscle. Our findings provide important and novel insights into the mechanisms underlying the beneficial effects of green light on goose embryos.

Transcriptome analysis reveals transforming growth factor-ß1 prevents extracellular matrix degradation and cell adhesion during the follicular-luteal transition in cows.

Ovarian angiogenesis is an extremely rapid process that occurs during the transition from follicle to corpus luteum (CL) and is crucial for reproduction. It is regulated by numerous factors including transforming growth factor-ß1 (TGFB1). However, the regulatory mechanism of TGFB1 in ovarian angiogenesis is not fully understood. To address this, in this study we obtained high-throughput transcriptome analysis (RNA-seq) data from bovine luteinizing follicular cells cultured in a system mimicking angiogenesis and treated with TGFB1, and identified 455 differentially expressed genes (DEGs). Quantitative real-time PCR results confirmed the differential expression patterns of the 12 selected genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified that the MAPK and ErbB pathways, cell adhesion molecules (CAMs), and extracellular matrix (ECM)-receptor interactions may play pivotal roles in TGFB1-mediated inhibition of CL angiogenesis. TGFB1 phosphorylated ERK1/2 (MAPK1/3) and Akt, indicating that these pathways may play an important role in the regulation of angiogenesis. Several genes with specific functions in cell adhesion and ECM degradation were identified among the DEGs. In particular, TGFB1-induced upregulation of syndecan-1 (SDC1) and collagen type I alpha 1 chain (COL1A1) expression may contribute to the deposition of type I collagen in luteinizing follicular cells. These results indicate that TGFB1 inhibits cell adhesion and ECM degradation processes involving ERK1/2, ErbB, and PI3K/Akt signaling pathways, and leads to inhibition of angiogenesis during the follicular-luteal transition. Our results further reveal the molecular mechanisms underlying the actions of TGFB1 in early luteinization.

Wider Angle Egg Turning during Incubation Enhances Yolk Utilization and Promotes Goose Embryo Development.

We aimed to investigate how wide-angle turning of eggs during incubation affected yolk utilization and the associated molecular mechanism, along with improved goose embryonic development. In total, 1152 eggs (mean weight: 143.33 ± 5.43 g) were divided equally and incubated in two commercial incubators with tray turning angles adjusted differently, to either 50° or 70°. Following incubation under the standard temperature and humidity level, turning eggs by 70° increased embryonic days 22 (E22), embryo mass, gosling weight at hatching, and egg hatchability, but reduced E22 yolk mass compared with those after turning eggs by 50°. Lipidomic analyses of the yolk revealed that egg turning at 70° reduced the concentrations of 17 of 1132 detected total lipids, including diglycerides, triglycerides, and phospholipids. Furthermore, the 70° egg turning upregulated the expression of genes related to lipolysis and fat digestion enzymes, such as lipase, cathepsin B, and prosaposin, as well as apolipoprotein B, apolipoprotein A4, very low-density lipoprotein receptor, low-density lipoprotein receptor-related protein 2, and thrombospondin receptor, which are genes involved in lipid transportation. Thus, a 70° egg turning angle during incubation enhances yolk utilization through the upregulation of lipolysis and fat digestion-related gene expression, thereby promoting embryonic development and improving egg hatchability and gosling quality.

Transforming growth factor-ß1 disrupts angiogenesis during the follicular-luteal transition through the Smad-serpin family E member 1 (SERPINE1)/serpin family B member 5 (SERPINB5) signalling pathway in the cow.

Intense angiogenesis is critical for the development of the corpus luteum and is tightly regulated by numerous factors. However, the exact role transforming growth factor-ß1 (TGFB1) plays during this follicular-luteal transition remains unclear. This study hypothesised that TGFB1, acting through TGFB receptor 1 (TGFBR1) and Smad2/3 signalling, would suppress angiogenesis during the follicular-luteal transition. Using a serum-free luteinising follicular angiogenesis culture system, TGFB1 (1 and 10ngmL-1 ) markedly disrupted the formation of capillary-like structures, reducing the endothelial cell network area and the number of branch points (P <0.001 compared with control). Furthermore, TGFB1 activated canonical Smad signalling and inhibited endothelial nitric oxide synthase (NOS3 ) mRNA expression, but upregulated latent TGFB-binding protein and TGFBR1 , serpin family E member 1 (SERPINE1 ) and serpin family B member 5 (SERPINB5 ) mRNA expression. SB431542, a TGFBR1 inhibitor, reversed the TGFB1-induced upregulation of SERPINE1 and SERPINB5 . In addition, TGFB1 reduced progesterone synthesis by decreasing the expression of steroidogenic acute regulatory protein (STAR ), cytochrome P450 family 11 subfamily A member 1 (CYP11A1 ) and 3ß-hydroxysteroid dehydrogenase (HSD3B1 ) expression. These results show that TGFB1 regulates NOS3 , SERPINE1 and SERPINB5 expression via TGFBR1 and Smad2/3 signalling and this could be the mechanism by which TGFB1 suppresses endothelial networks. Thereby, TGFB1 may provide critical homeostatic control of angiogenesis during the follicular-luteal transition. The findings of this study reveal the molecular mechanisms underlying the actions of TGFB1 in early luteinisation, which may lead to novel therapeutic strategies to reverse luteal inadequacy.

TNFα-Erk1/2 signaling pathway-regulated SerpinE1 and SerpinB2 are involved in lipopolysaccharide-induced porcine granulosa cell proliferation.

Lipopolysaccharide (LPS) is an inhibitory factor that causes hormonal imbalance and subsequently affects ovarian function and fertility in mammals. Previous studies have shown that the exposure of granulosa cells (GC) to LPS leads to steroidogenesis dysfunction. However, the effects of LPS on the viability of GC remain largely unclear. In the present study, we aimed to address this question and unveil the underlying molecular mechanisms using cultured porcine GC. Results showed that GC proliferation and tumor necrosis factor α (TNFα) secretion were significantly increased after exposure to LPS, and these effects were completely reversed by blocking the TNFα sheddase, ADAM17. Moreover, GC proliferation induced by LPS was mimicked by treatment with recombinant TNFα. In addition, SerpinE1 and SerpinB2 expression levels increased in GC after treatment with LPS or recombinant TNFα, whereas blocking the Erk1/2 pathway completely abolished these effects and also inhibited GC proliferation. Further, consistent with the effects of blocking the Erk1/2 pathway, cell proliferation was completely inhibited by knocking down SerpinE1 or SerpinB2 in the presence of LPS or recombinant TNFα. Mitochondrial membrane potential (MMP) polarization in GC was increased by LPS or recombinant TNFα treatment, and these changes were completely negated by Erk1/2 inhibition, but not by SerpinE1 or SerpinB2 knockdown. Taken together, these results suggested that the TNFα-mediated upregulation of SerpinE1 and SerpinB2, through activation of the Erk1/2 pathway plays a crucial role in LPS-stimulated GC proliferation, and the increase in GC MMP may synergistically influence this process.

Conception Rate and Reproductive Hormone Secretion in Holstein Cows Immunized against Inhibin and Subjected to the Ovsynch Protocol.

This study was conducted to investigate the feasibility of improving fertility in dairy cows via immunization against inhibin. Thirty-two cows were divided into Control (n = 11), Low-dose (n = 10) and High-dose (n = 11) groups. The High-dose and Low-dose cows were treated with 1 and 0.5 mg of the inhibin immunogen, respectively. All the cows were subjected to the Ovsynch protocol from the day of antigen administration and were artificially inseminated. Blood samples were serially collected over a 24-day period from the start of the Ovsynch protocol to 14 days after insemination. The results showed that immunization against inhibin dose-dependently increased the plasma concentrations of follicle-stimulating hormone (FSH), estradiol (E2), and activin A, but decreased progesterone (P4) concentrations in the luteal phase. Immunization also increased the plasma interferon (IFN)-τ concentrations in pregnant cows on day 14 after initial insemination. The conception rates in High-dose (45.5%) and Low-dose (40%) cows marginally increased compared to that in Control cows (27.3%), but the increases were not significant (p > 0.05). In conclusion, a single immunization against inhibin has the potential to improve conception rates, despite impaired luteal development. To further improve the reproductive performance of dairy cows, additional luteal-stimulating treatments are suggested in combination with immunization against inhibin and Ovsynch techniques.

ROS-Induced GATA4 and GATA6 Downregulation Inhibits StAR Expression in LPS-Treated Porcine Granulosa-Lutein Cells.

LPS is a major endotoxin produced by gram-negative bacteria, and exposure to it commonly occurs in animal husbandry. Previous studies have shown that LPS infection disturbs steroidogenesis, including progesterone production, and subsequently decreases animal reproductive performance. However, little information about the underlying mechanisms is available thus far. In the present study, an in vitro-luteinized porcine granulosa cell model was used to study the underlying molecular mechanisms of LPS treatment. We found that LPS significantly inhibits progesterone production and downregulates the expressions of progesterone synthesis-associated genes (StAR, CYP11A1, and 3ß-HSD). Furthermore, the levels of ROS were significantly increased in an LPS dose-dependent manner. Moreover, transcriptional factors GATA4 and GATA6, but not NR5A1, were significantly downregulated. Elimination of LPS-stimulated ROS by melatonin or vitamin C could restore the expressions of GATA4, GATA6, and StAR. In parallel, StAR expression was also inhibited by the knockdown of GATA4 and GATA6. Based on these data, we conclude that LPS impairs StAR expression via the ROS-induced downregulation of GATA4 and GATA6. Collectively, these findings provide new insights into the understanding of reproductive losses in animals suffering from bacterial infection and LPS exposure.

Corticosterone induces growth hormone expression in pituitary somatotrophs during goose embryonic development.

Treatment of fetal rat and embryonic chicken with exogenous glucocorticoids induces premature differentiation of growth hormone (GH) secreting cells. The effect of corticosterone (CORT) on somatotroph differentiation was mostly studied in pituitary cells in vitro. Currently, there is no evidence for glucocorticoid-mediated induction of somatotroph differentiation during pituitary development in bird species other than chicken. In this study, we sought to find out if in ovo injection of corticosterone into developing goose embryos could induce premature increase of GH in somatotrophs. On embryonic day (e) 15, the albumen of fertile goose eggs was injected with 300 µl of 0.9% saline, 300 µl 5 × 10-8M CORT, or 300 µl 5 × 10-6 M CORT. Embryos were allowed to develop until e20 and e28 and isolated pituitaries were subjected to quantitative real-time PCR and immunocytochemistry to detect GH mRNA and protein, respectively. At e20 and e28, blood from chorioallantoic vessels was subjected to radioimmunoassay for analysis of plasma GH protein. In ovo administration of exogenous corticosterone brought about a 2.5-fold increase in the expression of GH mRNA and increased the in situ expression of GH protein in goose pituitary cells, and enhanced plasma GH levels compared to that of the respective controls at e20. These findings prove that administration of glucocorticoid could stimulate the expression of GH in somatotrophs during goose embryonic development. Our results suggest the probable involvement of membrane glucocorticoid receptor in the corticosterone mediated expression of GH. Together with previously published data, our results suggest that corticosterone mediated induction of GH expression during embryonic development is relatively conserved among different vertebrates.

Expression and Purification of a Functional Porcine Soluble Triggering Receptor Expressed on Myeloid Cells 1.

Triggering receptor expressed on myeloid cells 1 (TREM-1) plays a vital role in the pathogen-triggered amplification loop required for proinflammatory responses. Blockade of TREM-1 signaling may inhibit expansion of sepsis and prolong survival of animals. In the present study, the gene of porcine soluble TREM-1 was cloned and expressed in E. coli. After purification, the bioactivity of recombinant porcine soluble TREM-1 was tested in vitro on porcine alveolar macrophages. The results showed that supplementation with the recombinant porcine sTREM-1 protein rapidly and dose-dependently attenuated the upregulation of cytokines (IL-1ß, IL-2, IL-4, IL-8, IL-10, IL-12, IL-16, IL-18, and TNF-α) caused by LPS stimulation in the cultured porcine alveolar macrophages. These results indicate that the recombinant porcine sTREM-1 protein can prevent TREM-1-mediated hyperinflammatory responses after exposure to LPS.

Investigating right ovary degeneration in chick embryos by transcriptome sequencing.

In asymmetric chick gonads, the left and right female gonads undergo distinct programs during development, generating a functional ovary on the left side only. Despite some progress being made in recent years, the mechanisms of molecular regulation remain incompletely understood, and little genomic information is available regarding the degeneration of the right ovary in the chick embryo testis. In this study, we performed transcriptome sequencing to investigate differentially expressed genes in the left and right ovaries and gene functions at two critical time points; embryonic days 6 (E6) and 10 (E10). Using high-throughput RNA-sequencing technologies, 539 and 1046 genes were identified as being significantly differentially expressed between 6R-VS-6L and 10R-VS-10L. Gene ontology analysis of the differentially expressed genes revealed enrichment in functional pathways. Among these, candidate genes associated with degeneration of the right ovary in the chick embryo were identified. Identification of a pathway involved in ovarian degeneration provides an important resource for the further study of its molecular mechanisms and functions.

Role of thymosin beta 4 in hair growth.

Although thymosin beta 4 (Tß4) is known to play a role in hair growth, its mechanism of action is unclear. We examined the levels of key genes in a Tß4 epidermal-specific over-expressing mouse model and Tß4 global knockout mouse model to explore how Tß4 affects hair growth. By depilation and histological examination of the skin, we confirmed the effect of Tß4 on hair growth, the number of hair shafts and hair follicle (HF) structure. The mRNA and protein expression of several genes involved in hair growth were detected by real-time PCR and western blotting, respectively. Changes in the expression of ß-catenin and Lef-1, the two key molecules in the Wnt signaling pathway, were similar to the changes observed in Tß4 expression. We also found that compared to the control mice, the mRNA and protein expression of MMP-2 and VEGF were increased in the Tß4 over-expressing mice, while the level of E-cadherin (E-cad) remained the same. Further, in the Tß4 global knockout mice, the mRNA and protein levels of MMP-2 and VEGF decreased dramatically and the level of E-cad was stable. Based on the above results, we believe that Tß4 may regulate the levels of VEGF and MMP-2 via the Wnt/ß-catenin/Lef-1 signaling pathway to influence the growth of blood vessels around HFs and to activate cell migration. Tß4 may have potential for the treatment of hair growth problems in adults, and its effects should be further confirmed in future studies.

Efficacy of progesterone supplementation during early pregnancy in cows: A meta-analysis.

Progesterone is a critical hormone during early pregnancy in the cow. As a result, a number of studies have investigated the effects of progesterone supplementation on pregnancy rates. In this study, a meta-analysis using a univariate binary random effects model was carried out on 84 specific treatments reported in 53 publications involving control (n = 9905) and progesterone-treated (n = 9135) cows. Although the results of individual studies showed wide variations (-40% to +50% point changes), progesterone treatment resulted in an overall increase in pregnancy rate odds ratio (OR = 1.12; P < 0.01). Improvements in pregnancy rate were only observed in cows treated at natural estrus (OR = 1.41, P < 0.01) and not following synchronization of estrus or ovulation. Although treatment between Days 3 to 7 postinsemination was beneficial (OR = 1.15; P < 0.01), treatment earlier or later than this was not. Progesterone supplementation was beneficial in cows of lower fertility (<45% control pregnancy rate) but not in cows with higher fertility. These results indicated that the benefit of progesterone supplementation on fertility of cows required exogenous progesterone supplementation to start between Day 3 to 7 and the appropriate reproductive status (i.e., lower fertility, natural estrus) of the treated cows.

Immunization against inhibin improves in vivo and in vitro embryo production.

The multiple ovulation and embryo transfer (MOET) technique has become an important breeding method in modern animal selection programs and a reproductive technique that can bypass ovarian dysfunction caused by heat stress to maintain reproductive performances in dairy cows. However, oocyte and embryo development often suffer from defects following repeated superovulation protocols. This phenomenon might be attributed to high levels of circulating inhibin, which is secreted by the supra-normal numbers of developing follicles during the process of superovulation. Through inhibin's negative impact on ovarian follicle development, high concentrations of inhibin might reduce oocyte quality and embryo developmental competence. Neutralizing endogenous inhibin bioactivities by active or passive immunizations against inhibin has been demonstrated to stimulate extra follicle development and induce multiple ovulations in both rodents and ruminants. Combined with conventional superovulatory protocols, immunization against inhibin further enhances follicle development and embryo yield. Furthermore, immunization against inhibin not only enhanced embryo quantity but also embryo quality in studies conducted in cows, sheep and water buffaloes. Similar beneficial effects on enhancing embryo development quality have been demonstrated in in vitro studies, where treatment with inhibin α subunit antibody enhances oocyte maturation and development of IVF or parthenogenically activated embryos. Thus, immunization against inhibin in combination with a conventional superovulation protocol can become a new technique to improve embryo production efficiency in vivo, as well as to develop a new oocyte IVM/IVF technique that can improve embryo IVP production efficiency.

Effects of dietary vitamin E on muscle vitamin E and fatty acid content in Aohan fine-wool sheep.

BACKGROUND: Increasing the polyunsaturated fatty acid (PUFA) content and decreasing the saturated fatty acid (SFA) content of mutton can help to improve its nutritional value for consumers. Several laboratories have evaluated the effects of vitamin E on the fatty acid (FA) composition of muscle in sheep. However, little information is available on wool sheep, even though wool sheep breeds are an important source of mutton, especially in northern China where sheep are extensively farmed. The present study was designed to address the effects of vitamin E on muscle FA composition in male Aohan fine-wool sheep. METHODS: Forty-two male Aohan fine-wool lambs (5 mo old) with similar initial body weight were randomly divided into seven groups and fed diets supplemented with 0 (control group), 20, 100, 200, 1,000, 2,000, or 2,400 IU/sheep/d vitamin E for 12 mo. Three lambs from each group were slaughtered to measure vitamin E and FA content in the longissimus lumborum (LL) and gluteus medius (GM) muscles. RESULTS: Vitamin E concentrations in the LL and GM increased significantly after 12 mo of vitamin E supplementation (P < 0.05). However, this increase did not occur in a dose-dependent manner because the muscle vitamin E concentration was highest in the 200 IU/sheep/d group. Dietary vitamin E supplementation also caused a significant reduction in SFA content and an increase in monounsaturated FA (MUFA) content in the LL and GM (P < 0.05). All six doses of vitamin E significantly increased cis9 trans11-conjugated linoleic acid (c9t11-CLA) content in the LL compared with the control group (P < 0.05). CONCLUSIONS: Dietary supplementation with vitamin E increased muscle vitamin E content and improved the nutritional value of mutton by decreasing SFA content and increasing MUFA and c9t11-CLA contents in Aohan fine-wool sheep. These effects were greatest in sheep fed a diet containing 200 IU/sheep/d vitamin E.

Molecular cloning and characterization of the sheep α-TTP gene and its expression in response to different vitamin E status.

The α-tocopherol transfer protein (α-TTP) is a ~32 kDa protein that exhibits a marked ligand specificity and selectively recognizes of α-tocopherol, which is the most active form of vitamin E. The α-TTP gene has been cloned and its physiological functions have been studied in numbers of species, however, the understanding of sheep α-TTP is still in his infancy. In this study, the full-length cDNA of sheep α-TTP gene was cloned from sheep liver by using of rapid amplification of complementary DNA ends (RACE). As a result, the sheep α-TTP gene was 1098 bp in nucleotide which contained 23 bp 5'-untranslated region (UTR), 226 bp 3'-UTR and 849 bp open reading frame (ORF) that encoded a basic protein of 282 amino acids. Further bioinformatic analysis indicated that the sheep α-TTP gene had a high homologous of both nucleotide and amino acid sequences compared with that of other species and had a Sec14p-like lipid-binding domain which called the CRAL-TRIO domain. Moreover, the expression of sheep α-TTP mRNA and protein in response to different vitamin E supplemented levels were observed according to quantitative real-time PCR (qRT-PCR) and Western blotting analysis. The results showed that dietary vitamin E levels did not affect α-TTP mRNA expression significantly while the low vitamin E supplemented level groups of sheep had significantly higher α-TTP protein compared to high-vitamin E groups.

Effect of Vitamin E supplementation on the enzymatic activity of selected markers in Aohan fine-wool sheep testis.

The aim of this study was to evaluate the effects of dietary Vitamin E supplementation on the testicular 'marker' enzyme activity and Vitamin E content in Aohan fine-wool sheep. Thirty male Aohan fine-wool sheep (5 months of age) with similar body weight were selected from the Aohan fine-wool sheep-breeding farm of Inner Mongolia Autonomous Region, China. The sheep were randomly divided into five groups and supplemented with 0, 20, 200, 1000 or 2400 IU sheep(-1)d(-1) Vitamin E for 12 months. Three sheep in each group were slaughtered at 17 months to collect a testis sample for testicular marker enzyme analysis. The results showed that, compared to Control, supplementing the diet with Vitamin E at 200 IU sheep(-1)d(-1) significantly increased the content of Vitamin E in testis and improved the activity of testicular mitochondrial ATPase (P<0.01), lactate dehydrogenase (LDH) (P<0.01), sorbitol dehydrogenase (SDH) (P<0.01), and alkaline phosphatase (ALP) (P<0.05). The present study demonstrated that supplementing Vitamin E can have a positive role in improving testicular marker enzyme activity and that the optimum range of dose appeared to be 100-200 IU sheep(-1)d(-1).

Effect of Vitamin E supplementation on semen quality and the testicular cell membranal and mitochondrial antioxidant abilities in Aohan fine-wool sheep.

Thirty male Aohan fine-wool sheep (5 months of age) with similar body weight were procured from the Aohan fine-wool sheep breeding farm of Inner Mongolia Autonomous Region, China. The sheep were divided randomly into five groups, which were labeled as Group 1, 2, 3, 4 and 5 and supplemented respectively with 0, 20, 200, 1000 or 2400IUsheep(-1)d(-1) Vitamin E, for 12 months, respectively. Three Aohan fine-wool sheep in each group were selected randomly for semen collection at the age of 16 months, then slaughtered at 17 months to collect the testis sample for testicular cell membranal and mitochondrial antioxidant abilities analysis. The results showed that supplementing Vitamin E at the concentration of 200IUsheep(-1)d(-1) in diets may have a positive effect in increasing semen quality and quantity (P<0.05), significantly reduce malondialdehyde (MDA) level and improve the activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-PX) in testicular cell membrane and mitochondria (P<0.05). In conclusion, the present study demonstrated that that supplementing Vitamin E can have a positive role in improving semen quality via protecting testicular cell membrane and mitochondria from antioxidant abilities. However, the optimal level of Vitamin supplement has still to be determined.