大鼠脂肪细胞分化过程中脂代谢相关基因转录表达时序的研究

对大鼠附睾和肾周脂肪组织分离的前体脂肪细胞原代培养，采用RT—PCR法分析脂代谢重要酶和转录因子基因在不同分化阶段转录表达的时序变化。结果显示，在前体脂肪细胞阶段，固醇调控元件结合蛋白（SREBP）-1c、碳水化合物反应元件结合蛋白（ChREBP）以及脂肪酸合成酶（FAS）、乙酰辅酶A羧化酶α（ACC1）、硬酯酰辅酶A去饱和酶（SCD）和激素敏感脂酶（HSL）基因均不转录表达；HSL mRNA在分化初期表达，中期表达水平最高，分化末期降低；SCD mRNA在分化中期表达，末期表达水平显著提高；分化初期FAS mRNA水平明显高于中期和末期；ACC1 mRNA仅在末期表达；分化初期SREBP-1c mRNA水平较高，中期和末期显著下降；ChREBP mRNA表达水平在分化末期稍高于中期，但差异不显著。表明，SREBP-1c mRNA的表达与脂肪细胞早期分化调控有关，分化成熟脂肪细胞的ChREBP mRNA表达可能与生脂基因的转录激活有关。     

溶血卵磷脂对肉仔鸡脂类代谢的影响

选200羽1日龄AA肉仔鸡,随机分成对照组和试验组(每组10个重复),试验组日粮添加0.1%溶血卵磷脂,研究溶血卵磷脂对肉鸡脂类代谢的影响。结果表明:溶血卵磷脂降低肝脏系数和肝脂率(P>0.05)、血清总甘油三酯与低密度脂蛋白胆固醇(LDL-C)(P>0.05)、LDL-C/HDL-C和葡萄糖含量(P<0.05),提高高密度脂蛋白胆固醇(HDL-C)和游离脂肪酸含量(P<0.05),上调肉仔鸡肝脏载脂蛋白B(P<0.01)和脂肪酸合成酶(P=0.155)mRNA表达水平;而对肉仔鸡的42日龄体重、腹脂率、皮下脂肪厚、肌间脂肪厚、腿肌和胸肌含脂率以及血清总胆固醇、三碘甲腺原氨酸、四碘甲腺原氨酸、促甲状腺激素刺激激素和胰岛素含量均无显著影响(P>0.05)。结果提示:溶血卵磷脂对肉仔鸡有减少肝脂肪沉积及降血脂作用。     

Effect of different types of litter material for rearing broilers

1. Coir dust was evaluated as broiler litter in comparison with sawdust and rice husk using 135 commercial broilers. Forty-five broiler chicks were reared to 42 d on a 50 mm layer of each of these litters. 2. Birds reared on coir dust showed no difference in food consumption, body weight gain, food conversion efficiency production number and survivability in comparison to those reared on saw dust and rice husk. 3. It was concluded that coir dust is suitable as broiler litter when cheaply available.     

Effect of delipidant agents during in vitro culture on the development,lipid content,gene expression and cryotolerance of bovine embryos

In vitro produced embryos are still sensitive to the freezing process which can be explained, in part, by the high-lipid accumulation that characterizes these embryos. Therefore, we aimed to evaluate the effect of delipidating agents, L-carnitine and the trans-10 cis-12 conjugated linoleic acid (CLA) isomer, on blastocyst development, lipid content, gene expression and cryotolerance when added to embryo culture media. Embryos were cultured in four different media: T1: control (n = 616), synthetic oviduct fluid (SOF) media with 5% foetal bovine serum (FBS); T2: L-carnitine (n = 648), SOF medium with 5% FBS and 0.6 mg/ml of L-carnitine; T3: CLA (n = 627), SOF medium with 5% FBS and 100 μM trans-10 cis-12 CLA; and T4: L-carnitine + CLA: (n = 597), SOF medium with 5% FBS plus 0.6 mg/ml L-carnitine and 100 μM trans-10 cis-12 CLA. Supplementation of culture medium with either or both delipidating agents reduced (p < .05) blastocyst rate on D7 (T1 = 49 ± 3.5; T2 = 39 ± 3.0; T3 = 42 ± 3.9 and T4 = 39 ± 3.9), but did not affected gene expression (p > .05). Although embryos cultured in the presence of L-carnitine contained fewer (p < .05) lipid droplets than the control embryos, they showed a lower re-expansion rate 24 hr post-thaw than those (p < .05). In conclusion, although L-carnitine reduced the amount of lipids in cultured embryos, the use of L-carnitine and CLA during in vitro culture was not able to improve the embryo production and the response to cryopreservation.     

Temporal expression of transforming growth factor-beta2 and myostatin mRNA during embryonic myogenesis in Indian broilers

TGF-beta2 and myostatin, the members of TGF family, act through both autocrine and paracrine mechanisms to regulate the growth and differentiation at various developmental stages in chicken. The kinetics and expression profile of these two growth factors were investigated by semi-quantitative RT-PCR, during the myogenesis of Indian broiler chickens. Total RNA was isolated from whole embryos on each of embryonic days (E) 0-6 (n=3 per day) and from the biceps femoris muscle at E7-E18 (n=3 per day). The expression of TGF-beta2 was noticed on E2 that remained at the same level until E6. In biceps femoris muscle, higher level of TGF-beta2 expression was observed during E7-E12, which decreased gradually thereafter. These findings suggested that TGF-beta2 might be a regulatory factor participating in the myogenesis of chicken embryos. Initial myostatin expression was noticed on E1, even before the myogenic lineage is established in embryo. This finding suggested an additional role of myostatin in early chicken embryo development, other than myogenesis. Furthermore, myostatin expression was significantly higher on E3 as compared to earlier studies, where initial higher level was observed at E2, suggesting the differential expression of myostatin among breeds. Higher and almost static myostatin expression was noticed in biceps femoris muscle during the entire period of myogenesis (E7-E18). In the present study, the ontogeny of myostatin expression coincided with myogenesis of chicken. Therefore, it may be hypothesized that myostatin is not only a major determinant of muscle mass, but also involved in early embryogenesis in chickens.     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background

Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring, 36 pregnant ewes were fed 60%, 100% or 140% of National Research Council requirements from d 31 ± 1.3 of gestation until parturition. Lambs from control-fed (CON), restricted-fed (RES) or over-fed (OVER) ewes were necropsied within 1 d of birth (n = 18) or maintained on a control diet for 3 mo (n = 15). Semitendinosus muscle was collected for immunohistochemistry, and protein and gene expression analysis.

Results

Compared with CON, muscle fiber cross-sectional area (CSA) increased in RES (58%) and OVER (47%) lambs at 1 d of age (P < 0.01); however at 3 mo, CSA decreased 15% and 17% compared with CON, respectively (P < 0.01). Compared with CON, muscle lipid content was increased in OVER (212.4%) and RES (92.5%) at d 1 (P < 0.0001). Muscle lipid content was increased 36.1% in OVER and decreased 23.6% in RES compared with CON at 3 mo (P < 0.0001). At d 1, myostatin mRNA abundance in whole muscle tended to be greater in OVER (P = 0.07) than CON. Follistatin mRNA abundance increased in OVER (P = 0.04) and tended to increase in RES (P = 0.06) compared with CON at d 1. However, there was no difference in myostatin or follistatin protein expression (P > 0.3). Phosphorylated Akt (ser473) was increased in RES at 3 mo compared with CON (P = 0.006).

Conclusions

In conclusion, maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring, possibly through altered gene and protein expression.     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background:Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth.To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring,36 pregnant ewes were fed 60%,100%or 140%of National Research Council requirements from d 31 ± 1.3 of gestation until parturition.Lambs from control-fed(CON),restricted-fed(RES) or over-fed(OVER) ewes were necropsied within 1 d of birth(n = 18) or maintained on a control diet for 3 mo(n = 15).Semitendinosus muscle was collected for immunohistochemistry,and protein and gene expression analysis.Results:Compared with CON,muscle fiber cross-sectional area(CSA) increased in RES(58%) and OVER(47%)lambs at 1 d of age(P 0.01);however at 3 mo,CSA decreased 15%and 17%compared with CON,respectively(P 0.01).Compared with CON,muscle lipid content was increased in OVER(212.4%) and RES(92.5%) at d 1(P 0.0001).Muscle lipid content was increased 36.1%in OVER and decreased 23.6%in RES compared with CON at 3mo(P 0.0001).At d 1,myostatin mRNA abundance in whole muscle tended to be greater in OVER(P = 0.07) than CON.Follistatin mRNA abundance increased in OVER(P = 0.04) and tended to increase in RES(P = 0.06) compared with CON at d 1.However,there was no difference in myostatin or follistatin protein expression(P 0.3).Phosphorylated Akt(ser473) was increased in RES at 3 mo compared with CON(P = 0.006).Conclusions:In conclusion,maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring,possibly through altered gene and protein expression.     

Gene expression profiling in embryonic chicken ovary during asymmetric development

The reproductive system in female birds arises as bilateral asymmetrical anlagen, excluding the birds of prey. Earlier, histological and messenger RNA (mRNA) expression profile studies of several genes related to gonadal sex differentiation in chicken embryos tried to elucidate the query of this asymmetry in a scattered manner. To understand the matter precisely, we have focused on mRNA expression of a cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) in second half of the embryonic days (E10–E18). The established role of leptin in development of the embryo and its expression in the embryonic ovary also drove us to check leptin receptor (LEPR) expression in the ovary. Increased expression of FSHR and CYP19A1 in the left ovary compared with that in the right ovary was identified (P < 0.05), promoting preferential left ovarian development and functionality. Significant high expression (P < 0.05) of the apoptotic genes in the right ovary were also involved here. Leptin probably has no direct influence on ovarian asymmetry as no significant variation in gonadal mRNA expression of LEPR was observed within the same experimental days. We propose that asymmetric expression of this cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) leads to the development of dimorphic gonads during embryogenesis.     

母体营养对后代肌肉生长发育的影响

骨骼肌占机体干重的50%以上,是机体的重要组成部分。肌肉的发育受很多因素的调控,而在关键窗口期,母体营养对其影响是至关重要的。母体营养状况可以改变后代肌纤维的数目、密度、肌肉的代谢、胰岛素敏感性和肌内脂肪的沉积。本文就母体营养对后代肌肉发育的影响作一简要概述。     

Effect of intravenously injected zinc on tissue zinc and metallothionein gene expression of broilers

1. The effect of intravenously injected zinc (Zn) on tissue Zn concentrations and pancreas metallothionein (MT) gene expression in broilers was investigated to detect differences in the tissue utilisation of Zn from different Zn sources.

2. A total of 432 male chickens were randomly allotted on d 22 post-hatch to one of nine treatments in a completely randomised design. Chickens were injected with either a 0.9% (w/v) NaCl solution (control) or a saline solution supplemented with Zn sulphate or one of three organic Zn chelates with weak (Zn-AA W), moderate (Zn-Pro M) or strong (Zn-Pro S) chelation strengths at two injected Zn dosages calculated according to two Zn absorbability levels (6 and 12%).

3. Bone and pancreas Zn concentrations, pancreas MT mRNA levels and MT concentrations increased on d 6 and 12 after Zn injections as the injected Zn dosages increased. Chickens injected with the Zn-Pro S had lower bone Zn concentration than those injected with the Zn-Pro M or Zn-AA W on d 6 after injections. However, no differences among Zn sources were observed in bone Zn concentration on d 12 after injections, pancreas Zn concentrations, pancreas MT mRNA levels and MT concentrations on both d 6 and d 12 after injections.

4. It was concluded that the injected Zn-Pro S was the least favourable for bone Zn utilisation of broilers. The pancreas Zn concentration and pancreas MT gene expressions might not be sensitive enough to detect differences in the tissue utilisation of injected Zn in broilers between organic and inorganic Zn sources or among organic Zn sources.     

Effects of contact dermatitis on hepatic gene expression in broilers

1.?Severe foot and hock dermatitis in broiler chickens can reduce growth rate and increase susceptibility to bacterial infection, affecting both profitability and welfare. However, little is known about the underlying physiological changes associated with foot and hock lesions.

2.?This study compared global hepatic gene expression in control birds and those with ammonia-induced foot and hock lesions using Agilent 44?K chicken oligonucleotide microarrays (8 birds per group).

3.?In total, 417 genes were differentially expressed of which 174 could be mapped onto the genome. Genes associated with energy metabolism, thyroid hormone activity and cellular control were affected, while there was also evidence of an up-regulation of genes linked to a pro-inflammatory response.

4.?It is conceivable that pain is the underlying cause for the observed changes in energy metabolism genes.

5.?Changes in hepatic gene expression provide new information on how a chicken's physiological mechanisms alter to cope with foot and hock lesions. The findings support other data indicating that birds with increased severity of lesions are likely to be in pain and that growth will be compromised. Reduction of the incidence of dermatitis by improved husbandry should therefore benefit both welfare and commercial performance.     

马杜霉素对肉仔鸡免疫器官发育影响的研究

为了探讨马杜霉素对肉仔鸡免疫器官的影响,试验选择体重相近的1日龄肉仔鸡200只,在日粮中分别不添加马杜霉素(Ⅰ组)和添加5.0 mg/kg(Ⅱ组)、7.5 mg/kg(Ⅲ组)、10.0 mg/kg(Ⅳ组)的马杜霉素,连续饲喂4周,测定各组雏鸡胸腺、脾脏、法氏囊指数。结果表明:5.0 mg/kg马杜霉素3周对肉仔鸡免疫器官发育有一定促进作用,但3周后呈现一定的抑制作用;用7.5 mg/kg和10.0 mg/kg马杜霉素饲喂1周即可抑制免疫器官的生长发育,其抑制作用能持续至停药后1周,但在停药3周后基本恢复正常。说明马杜霉素对肉仔鸡免疫器官发育的抑制作用呈现一定的量效和时效效应。     

Effect of copper deficiency on blood lipid profile and haematological parameters in broilers

The aim of the study was to observe the changes in haematological parameters and blood lipid profile through copper (Cu) deficiency. Eighty broiler chicks were used in the study. The chicks were separated into two equal groups (n = 40), feed treatment according to the groups started on day 7. The animals in the control group were fed with normal feed. Those in the Cu-deficient group were fed with feed that did not contain supplemental Cu. Blood samples were taken from all the animals on days 7, 21 and 49. Fibrinogen and prothrombin time were assayed in plasma; total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol (LDL-cholesterol), triglyceride and ceruloplasmin were assayed in sera; and haematocrit and thrombocyte levels were assayed in the blood samples. Total cholesterol, triglyceride and LDL-cholesterol levels were significantly higher in the Cu-deficient group than in the control group. Serum ceruloplasmin levels were lower in the Cu-deficient group than in the control group. Prothrombin time was higher in the Cu-deficient group than in the control group. Haematocrit levels were lower in the Cu-deficient group than in the control group. It was concluded that Cu deficiency created hypertriglyceridaemia, hypercholesterolaemia, insufficient growth and anaemia in broilers.     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Effect of number of conceptuses on maternal mammary development during pregnancy in the pig

An experiment was conducted to determine if manipulation of conceptus number could alter estrogen, progesterone or prolactin concentrations in gilts and thereby influence mammary development. Estradiol-valerate injections, oviduct ligation or no treatment were utilized to form three groups of gilts with zero (n=5), 4 to 7 (n=4), or 8 to 11 (n=4) conceptuses, respectively. Blood samples were taken throughout pregnancy, and at day 110 of gestation all gilts were slaughtered and total mastectomies were performed. Mammary glands were analyzed for wet weights, dry weights, RNA and DNA contents. Results indicated conceptuses stimulated mammary development since mammary glands from pseudopregnant gilts had only 22% as much DNA as mammae from pregnant gilts. However, mammary DNA was not different between gilts with 4 to 7 or 8 to 11 conceptuses. The relationship between conceptus number and mammary gland DNA in gilts was best described by the quadratic equation y = 577 + 583x − 38x², where y = total mammary DNA (mg) and x = conceptus number. Significant positive among-animal and within-group correlations between estrogen-sulfate or estrone concentrations in blood and maternal mammary DNA content suggest that estrogens may be the primary conceptus-derived mammogenic hormone in gilts.     

高温与高密度对黄羽肉鸡血液生化指标和免疫机能的影响

试验选用 2 8日龄健康快大岭南黄羽肉鸡 ,采用 2 (环境温度 )× 2 (饲养密度 )因子随机分组设计 ,根据体重和性别一致原则随机分为 4组 ,每组 4个重复 ,试验饲粮各组一致。试验期 42d。结果表明 :高温与高密度对试验鸡血液生化指标、免疫指标及免疫器官重与比例具有不同程度的影响 ,而且两者对试验鸡血清Na+ 、尿酸氮、总蛋白、白蛋白含量、白蛋白 /球蛋白比值、CD8+ 含量和法氏囊重及其比例影响存在显著或极显著互作。     

Hormone-mediated maternal stress affects embryonic development during incubation without adverse effect on chick weight and body composition

1. The objective of this study was to evaluate the effect of maternal stress (MS) induced by supplementing the hen’s diet with 2 mg/hen/d dietary corticosterone (CORT) on embryonic development, biochemical blood parameters and hatching performance of broiler chicks.

2. A total of 200 Ross broiler breeder hens at 42 weeks of age were randomly divided into two groups: MS or control. Hens in the MS were fed 2 mg/hen/d CORT for 14 d. Eggs (648 and 635 eggs for MS and control, respectively) were collected from d 3 to 14 of dietary CORT supplementation and incubated. Weights of embryo, chicks and organs and body composition were determined during incubation and at hatch. Biochemical blood parameters were measured at internal pipping stage and day of hatch. Hatching performance and embryonic mortalities were recorded.

3. Hens fed a diet supplemented with CORT had lighter body weight and produced less eggs at the end of the 14-d treatment period. Although MS embryos were heavier than control from 12 to 18 d of incubation, chick weight was similar at the day of hatch. Lower relative weights for yolk sac and bursa were observed at 12 d of incubation for MS chicks compared to control. Chicks from both groups had similar body content in spite of higher fat content of MS embryos on d 18 of incubation.

4. MS had no effect on the duration of incubation or hatching performance but increased mortality at the pipping stage.

5. The results suggest that hormone-mediated MS might affect embryonic development during incubation without adverse effect on chick weight and body composition.     

谷氨酰胺对肉仔鸡脾组织结构发育的影响

160只 1日龄艾维茵肉仔鸡随机分成 4组 ,分别饲喂添加 0 % ,0 2 % ,0 4%和 0 8%谷氨酰胺的饲粮 2 8d。每周末每组取 6只鸡 ,颈静脉放血致死 ,取脾脏 ,Bouin液固定 ,制作石蜡切片 ,HE染色 ,光镜观察 ,显微摄影 ,研究基础日粮中添加谷氨酰胺对肉仔鸡脾组织结构的影响。结果显示 :添加谷氨酰胺的各试验组脾小结增多、增大 ;动脉周围淋巴鞘增厚 ;椭球增多、增大 ,细胞排列疏松 ,鞘毛细血管内皮细胞增高、增多 ;红髓比例降低 ,脾索内浆细胞和巨噬细胞增多。添加 0 8%谷氨酰胺的脾小结变化明显 ,添加 0 4%谷氨酰胺的动脉周围淋巴鞘和椭球变化明显。说明日粮中适量添加谷氨酰胺可促进脾内淋巴细胞的增殖与分化 ,从组织学角度证明 ,谷氨酰胺可促进脾的免疫应答能力 ,提高机体的免疫功能     

Dynamics of apoptosis-related gene expression during follicular development in yak

The potential reproduction power of domestic animals is limited by a complicated follicular atresia process. P53, caspase-9 (Casp9), Bax, Bcl-2 and Fas play a crucial role in the ovarian mitochondrion-dependent apoptosis and death receptor pathway. In accordance with this study, the expression levels of Casp9, Bax, Bcl-2 and Fas were analysed in ovaries and oviducts of yak by immunohistochemistry (IHC). P53 and the above in ovarian granulosa cells (GCs) from atretic (3–6 mm) to healthy follicles (6–8 mm) and in oviducts were examined from the luteal phase to the follicular phase during the oestrous circle by Western blot (WB) and real-time PCR (RT-PCR). Results demonstrated that typical classic apoptotic factors Casp9, Bax, Bcl-2 and Fas were expressed in the cytoplasm and zonal pellucida of oocytes, primordial follicles, primary follicles, ovarian surface epithelium, ovarian GCs, granular lutein cells, surface epithelia in oviduct uterotubal junction and oviduct ampulla during the luteal phase. RT-PCR and WB revealed that P53 and Fas significantly increased in GCs of atretic follicles. P53 and Casp9 increased in oviduct epithelium during the luteal phase, but Fas was unchanged. A contrary tendency was noted in Bcl-2 and Bax expression. Overall, P53 and Fas play an essential role in inducing GC apoptosis, and Bax, Bcl-2, Casp9 and P53 are involved in oviduct epithelial regeneration in yak.