lncLER在卢氏绿壳蛋鸡肝脏中表达调控的研究

长链非编码RNA（lncRNA）在许多生物学过程中发挥着重要的调节作用。河南农业大学家禽种质资源创新工程研究中心通过对产蛋前期（20周龄）和产蛋高峰期（30周龄）卢氏绿壳蛋母鸡肝脏组织的转录组分析,筛选出了一个在产蛋高峰期母鸡肝脏中表达水平显著上调的lncRNA（lncLER）。本研究利用半定量PCR（RT-PCR）、实时荧光定量PCR分析了lncLER在卢氏绿壳蛋母鸡不同发育时期和不同组织中的表达规律,并从个体和细胞水平研究了体内、体外雌激素处理对lncLER表达的影响。结果表明,lncLER在卢氏绿壳蛋母鸡各组织中均有不同程度的表达,且在产蛋高峰期肝脏中的表达量较高;雌激素在个体和细胞水平可显著上调lncLER的表达。本试验初步阐明了lncLER基因在卢氏绿壳蛋母鸡中的表达调控规律,为进一步研究lncLER在鸡产蛋过程中的生物学功能奠定了基础。     

鸡微粒体甘油三酯转运蛋白样基因生物学特性及表达调控

旨在探究鸡微粒体甘油三酯转运蛋白样基因（MTTPL）生物学特性及其表达调控机制,为进一步探讨其在鸡肝脂质代谢中的生物学功能奠定基础。本研究首先采用PCR和测序技术,克隆MTTPL cDNA序列;利用在线软件对MTTPL蛋白结构域、三维空间结构及系统进化树进行分析;构建pcDNA3.1-MTTPL-EGFP过表达载体,通过与内质网标签蛋白的表达载体DsRed-λ共转染鸡肝癌细胞系（LMH）细胞,对MTTPL进行亚细胞定位;分别采集不同周龄卢氏绿壳蛋鸡（1日龄、1周龄、10周龄、30周龄每个周龄各8只）组织样,采用荧光定量PCR分析MTTPL基因和鸡微粒体甘油三酯转运蛋白基因（MTTP）的时空表达谱;然后用0.5、1和2 mg·kg^-1体重浓度的17 β-雌二醇分别处理海兰褐鸡（每组20只）12和24 h后,分析雌激素对肝MTTPL和MTTP表达的影响;最后用1、50、和100 nmol·L^-1 17β-雌二醇及雌激素受体拮抗剂分别处理鸡胚肝原代细胞12 h （每组3个重复）,研究雌激素调控MTTPL基因表达的作用机制。结果表明,鸡MTTPL的CDS区全长为2 646 bp,可编码881个氨基酸,与人和鸡MTTP拥有共同的祖先;定位于细胞内质网;鸡MTTPL和MTTP具有与人MTTP相同的功能域,且三维结构相似度偏差较小,分别为0.090和0.064;MTTPL基因在鸡肝和肾组织中高表达,MTTP在鸡肝、肾和小肠中高表达;在肝中,MTTPL和ApoB的表达水平随周龄的增加均显著上升（P<0.05）,而MTTP的表达仅在10周前随周龄增加显著升高（P<0.05）,而在产蛋前（10周龄）和产蛋期（30周龄）无显著变化（P>0.05）;在17β-雌二醇处理鸡12和24 h时后,MTTPL及ApoB在肝中均显著上调表达（P<0.05）,而MTTP在高浓度处理时表达量显著下调（P<0.05）,在低浓度时无显著变化;与对照组相比,雌激素可显著上调鸡胚肝原代细胞中ApoB和MTTPL的表达（P<0.05）,而MTTP表达水平无显著变化;与雌激素处理组相比,雌激素及受体ERα拮抗剂MPP共处理组MTTPL基因表达水平显著降低（P<0.05）;与MPP和雌激素共处理组相比,ERα和ERβ受体拮抗剂ICI和TAM处理组MTTPL基因表达水平无显著变化。综上所述,鸡MTTPL位于内质网中,具有与人MTTP相似的功能结构域;MTTPL和MTTP均在肝中相对高表达,且MTTPL在产蛋期鸡肝的表达显著高于产蛋前期,而MTTP无显著变化。雌激素可通过与ERα受体结合调控鸡MTTPL的表达,而MTTP不受雌激素调控。表明MTTPL可能在鸡产蛋期肝脂质代谢中发挥重要作用。     

QDPR基因克隆及其在乌蒙凤鸡不同生长阶段组织表达研究

旨在克隆乌蒙凤鸡醌型二氢生物喋呤还原酶（quinoid dihydropteridine reductase，QDPR）基因，并检测其在不同性别乌蒙凤鸡不同生长阶段各组织中的表达差异，以研究QDPR基因在乌蒙凤鸡生长发育过程中的调控作用。本试验对乌蒙凤鸡QDPR基因CDS区进行PCR扩增和克隆，并对得到的序列进行生物信息学分析，通过实时荧光定量PCR检测QDPR基因在4、12、20和28周龄乌蒙凤鸡公鸡及母鸡的心、肝、脾、肺、肾、胸肌和腿肌组织中的相对表达量。结果表明，QDPR基因开放阅读框长度为417 bp，共编码139个氨基酸，编码蛋白为酸性不稳定蛋白，乌蒙凤鸡的QDPR基因序列与原鸡、日本鹌鹑和珠鸡的QDPR基因同源性较高；QDPR基因在不同性别乌蒙凤鸡不同生长阶段各组织中均有表达，其中，公鸡4、12和20周龄肺组织中QDPR基因表达量最高，极显著高于同一周龄其他组织（P<0.01），母鸡4周龄肝、脾、肺中QDPR基因相对表达量为所有时期最高，极显著高于12、20周龄（P<0.01），公鸡大多数组织中QDPR基因表达量随时间增长总体呈现出先升后降的趋势，母鸡为先降后升。结果显示，QDPR基因在乌蒙凤鸡内脏组织中的表达均高于肌肉组织，且公母鸡不同时期表达情况有所差异，试验结果可为进一步研究QDPR基因在鸡生长发育中的调控作用提供参考。     

乙型脑炎病毒感染PK15细胞的lncRNA差异表达谱分析

旨在分析乙型脑炎病毒(Japanese encephalitis virus,JEV)感染猪肾上皮细胞PK15后的lncRNA差异表达谱,探究宿主lncRNA在JEV感染和宿主防御中的潜在功能.本研究中利用JEV感染PK15细胞,感染36 h,收集细胞,同时设立未感染对照组,每组3个生物学重复.提取细胞总RNA,构建c...     

大豆黄酮对伊莎鸡卵泡发育及其P450arom基因表达的影响

实验探讨了大豆黄酮(DAI)对伊莎鸡卵泡发育及其芳香化酶(P450arom)mRNA表达的影响。实验选取16只产蛋后期伊莎鸡,等分为对照组和DAI处理组。对照组饲喂基础日粮,实验组在基础日粮中添加10 mg/kgDAI。实验持续7周后,分离排卵前卵泡(F1、F2、F3……)的颗粒层及小黄卵泡和大白卵泡,通过RT-PCR法检测P450arom mRNA表达的相对丰度。结果表明:DAI明显提高了伊莎鸡小黄卵泡和大白卵泡的数量,P450arommRNA在伊莎鸡不同发育阶段卵泡中的表达存在差异,部分卵泡P450arom mRNA表达的相对丰度显著增加。因此,在产蛋后期伊莎鸡基础日粮中添加DAI可增加不同发育阶段卵泡的数目,上调部分卵泡中与发育相关的基因表达以促进卵泡发育。     

钙蛋白酶基因在鸡不同组织和品种中的表达差异

为了探讨钙蛋白酶(calpain,CAPN)基因在鸡不同组织和品种中的表达差异,本研究采用半定量RT-PCR方法研究了CAPN基因在鸡不同组织和品种中的表达差异。结果表明:CAPN基因表达于优质鸡的10个不同组织中,肝脏和胸肌中的表达量最高并显著高于其他组织(P<0.05);优质鸡胸肌组织中CAPN基因的表达量显著高于艾维茵肉鸡和宝万斯尼拉蛋鸡品种(P<0.05),优质鸡品种之间以及肉鸡与蛋鸡品种间差异不显著(P>0.05)。结果证明钙蛋白酶是一种在鸡体内普遍存在的蛋白,CAPN基因在鸡不同组织和品种中的表达存在显著差异。     

产蛋前期和产蛋高峰期鸡全基因组甲基化差异及其对肝脏转录组影响的研究

旨在通过对产蛋前期和产蛋高峰期鸡肝全基因组甲基化差异进行分析,解析基因组甲基化对不同发育阶段肝中基因表达差异的影响。本研究采用全基因组重亚硫酸盐测序（WGBS）技术对产蛋前期（20周龄）和产蛋高峰期（30周龄,各3只DNA混池）卢氏绿壳蛋鸡肝全基因组的甲基化水平进行检测,并与已有的肝mRNA转录组数据进行整合分析,探讨基因组甲基化对不同生理阶段基因表达差异的影响。结果表明,全基因组范围约有4%的胞嘧啶（C）发生了甲基化（mC）;两个生理阶段的总体甲基化水平基本一致。共检测到670个差异甲基化区域（DMRs）和356个差异甲基化基因（DMGs）。基因本体（GO）和相关信号通路（KEGG）分析发现,超甲基化DMGs显著富集在发育的正向调控、细胞形态改变的调控、VEGF信号通路、肌动蛋白细胞骨架的调控、粘着斑及间隙连接等相关过程,低甲基化DMGs显著富集在胚胎消化道形态的发生、间充质细胞增殖的正向调控、淀粉和蔗糖代谢及Wnt信号通路等相关过程。基因不同功能区域甲基化水平与基因表达水平有关,启动子（promoter）及基因体（gene body）区域甲基化水平与基因表达水平呈显著负相关,其他区域（内含子、3'UTR）的甲基化水平与基因的表达水平无明显关系。其中,与肝脂质代谢相关的候选基因RASD1、HAO1、UBE2O、MSRB3受甲基化调控。本研究绘制了不同生理时期卢氏绿壳蛋鸡全基因组甲基化图谱,结合mRNA转录组数据阐述了DNA甲基化在基因表达方面的调控作用,并鉴定出了不同生理时期受甲基化调控的基因,为深入研究表观遗传调控在不同生理时期蛋鸡肝代谢中的作用机制提供参考。     

海兰褐蛋鸡ZP3基因结构及其在卵泡中表达规律分析

旨在探讨ZP3基因在海兰褐蛋鸡中的表达特性，为后期研究ZP3基因在蛋鸡卵泡发育中的作用提供理论支撑。本研究以产蛋期50周龄（50 W）海兰褐蛋鸡为试验对象，利用RACE技术克隆ZP3基因全长，并对编码区进行生物信息学分析。选择健康、体重相近的海兰褐蛋鸡6只，分别取心、肝、肺、肾、卵巢、胸肌、腿肌和不同等级卵泡构建表达谱。卵巢颗粒细胞经不同浓度（PBS、5 ng·mL^-1、10 ng·mL^-1、20 ng·mL^-1）促卵泡素（follicle-stimulating hormone，FSH）处理后，提取总RNA，采用实时荧光定量检测ZP3基因在各个样品的表达水平。结果表明，鸡ZP3基因全长1 415 bp，其中编码区1 341 bp，共编码446个氨基酸，位于10号染色体上，包含9个外显子；其亲缘关系与猪最远；跨膜结构预测表示，其含有一个跨膜结构域和一个信号肽区域；对其亲疏水性进行分析，预测该蛋白为弱的疏水性蛋白，蛋白质结构主要由无规则卷曲构成。组织表达谱显示，ZP3基因在鸡的卵巢中相对表达量最高。各等级卵泡ZP3表达分析显示，随着卵泡发育，ZP3基因在成熟卵泡（F1）颗粒细胞层表达量最高；在使用FSH处理等级颗粒细胞后，发现ZP3表达量显著上调（P<0.05），暗示ZP3基因在颗粒细胞中受到FSH激素调节。本试验对ZP3基因的结构进行了分析并预测该基因存在跨膜结构和信号肽区域。基因表达谱分析结果显示，ZP3基因在卵泡发育过程中表达量逐渐升高且主要在颗粒细胞中表达，可能受到FSH的调节作用，因此预测，ZP3基因可能与海兰褐蛋鸡繁殖功能相关。     

Differential expression of peroxisome proliferator-activated receptors alpha and gamma gene in various chicken tissues

The peroxisome proliferator-activated receptors (PPARs) are the members of superfamily of nuclear hormone receptors. A great number of studies in rodent and human have shown that PPARs were involved in the lipids metabolism. The goal of the current study was to investigate the expression pattern of PPAR genes in various tissues of chicken. The tissue samples (heart, liver, spleen, lung, kidney, stomach, intestine, brain, breast muscle and adipose) were collected from six Arber Acres broilers (8 weeks old, male and female birds are half and half). Semi-quantitative RT-PCR and Northern blot were used to characterize the expression of PPAR-alpha and PPAR-gamma genes in the above tissues. By semi-quantitative RT-PCR, the results showed the expression level of PPAR-alpha gene was higher in brain, lung, kidney, heart and intestine, medium in stomach, liver and adipose than in spleen, and it did not express in breast muscle. The expression level of PPAR-gamma gene was higher in adipose, medium in brain and kidney than in spleen, heart, lung, stomach and intestine, but it did not express in liver and breast muscle. Northern blot results showed that PPAR-alpha gene expressed in heart, liver, kidney and stomach, and the intensity of hybridization signal was the stronger in liver and kidney than in other tissues, however, PPAR-gamma gene only expressed in adipose and kidney tissues. The results of this study showed the profile of PPAR gene expression in the chicken was similar to that in rodent, human and pig. However the expression profile of chicken also have its own specific trait, i.e. compared with mammals, PPAR-alpha gene can not be detected in skeletal muscle and PPAR-gamma gene can be stronger expressed in kidney tissues. This work will provide some basic data for the PPAR genes expression and lipids metabolism of birds.     

鸡lncRNA-MSTRG.15568.9及其预测靶基因的表达

试验旨在了解在鸡睾丸中高表达的1个长链非编码RNA （long non-coding RNA,lncRNA）及其预测靶基因的时空表达规律,研究二者在鸡弱精子症中的调控作用。根据弱精子症和正常北京油鸡公鸡睾丸转录组测序筛选到的1个高表达的lncRNA （MSTRG.15568.9）,采用顺式（cis）作用模式预测其潜在靶基因SPAG4（sperm-associated antigen 4）,进一步采用实时荧光定量PCR方法进行表达量分析。分别选择3只0、5、20、30、45、60周龄正常北京油鸡公鸡,检测MSTRG.15568.9与SPAG4基因在不同周龄公鸡睾丸中的表达量差异;选择30周龄3只正常公鸡,采集睾丸、肝脏和脾脏等8个部位组织样品,检测MSTRG.15568.9与SPAG4基因在不同组织间的表达规律;选择45周龄弱精子症公鸡和正常公鸡各3只,对比MSTRG.15568.9与SPAG4基因在睾丸的表达量差异。结果显示,MSTRG.15568.9与SPAG4存在明显的时空表达差异,且二者表达趋势基本一致。在不同周龄的鸡睾丸组织中,MSTRG.15568.9和SPAG4的表达趋势相近,MSTRG.15568.9在20周龄的表达量显著高于0、5、30、45、60周龄（P<0.05）,0和5周龄表达量显著低于20、30、45和60周龄（P<0.05）;SPAG4在45周龄表达量最高,其次是20周龄（P<0.05）。MSTRG.15568.9和SPAG4在睾丸和肝脏中的表达量均显著高于脾脏、肾脏等组织（P<0.05）;在正常睾丸组织中的表达量均显著高于弱精子症睾丸组织（P<0.05）。综上所述,MSTRG.15568.9与SPAG4基因具有较明显的组织表达特异性,且MSTRG.15568.9可能调控SPAG4基因的表达,参与精子发生与精子活力调控;但其具体作用机制需要进一步探索。本研究可为鉴定与鸡弱精子症调节机制相关的功能基因提供参考。     

野生盘羊×巴什拜羊杂交二代尾部脂肪差异表达lncRNA的鉴定与分析

【目的】 分析不同脂尾型绵羊尾部脂肪组织中的长链非编码RNA(long non-coding RNA,lncRNA)的表达谱,探究lncRNA与绵羊尾部脂肪沉积机制的关系,为揭示绵羊尾脂沉积机制提供理论依据。【方法】 选择新疆地方绵羊巴什拜羊(肥尾型)和野生盘羊×巴什拜羊杂交二代(小尾型)作为研究对象,采集2个群体尾部脂肪组织,利用转录组测序技术筛选差异表达lncRNA并预测靶基因,对其进行GO功能和KEGG通路富集分析;利用实时荧光定量PCR技术验证转录组测序的表达结果。【结果】 通过差异表达分析共筛选出728个差异表达lncRNAs,其中270个表达下调,458个表达上调;通过差异表达lncRNA靶基因的GO功能和KEGG通路富集分析,筛选到634个靶基因参与疾病、免疫、蛋白修饰、细胞代谢等相关功能分类,共涉及76条信号通路,部分lncRNA介导的靶基因SCD、GPAM、THRSP、FASN等与绵羊尾部脂肪沉积相关。实时荧光定量PCR与转录组测序结果趋势相同。【结论】 lncRNA在绵羊脂尾进化中对尾部脂肪沉积具有重要的调控作用,为从lncRNA的角度分析绵羊脂肪沉积提供了理论依据。     

Ascorbic acid synthesis and transportation capacity in old laying hens and the effects of dietary supplementation with ascorbic acid

Background: Laying hens over 75 weeks of age commonly show great declines in immunity and production performance.It is unclear whether these declines can be relieved by supplementing with ascorbic acid(AA) in feed.Two trials were conducted to investigate the synthesis and metabolism of AA in layers of different ages and the effects of dietary supplemental AA on the performance and the immune and antioxidant statuses of 78 weeks old hens.Methods: In Exp.1,equal numbers(24 hens) of 35 weeks old(Young) and 75 weeks old(Old) layers were fed the same diet without AA supplementation for 4 weeks.In Exp.2,360 healthy 78 weeks old laying hens were randomly assigned to 4 treatments(basal diet supplemented with 0,0.25,0.5,or 1 g AA/kg diet) in an 8-week feeding trial.Results: The old hens tended to have decreased L-gulonolactone oxidase(GLO) synthase activity in the kidney and liver than that of the young hens(P = 0.07 and P = 0.05,respectively).Compared with the young hens,the old hens had lower hepatic antioxidant capacity allowing for the lower thioredoxin(TXN),thioredoxin reductase(TXNR) and cytochrome b5 reductase(CYB5 R) gene expression(P 0.05),whereas increased sodium-dependent vitamin C transporter(SVCT) 1 expression levels in the ileum and kidney and enhanced splenic and hepatic AA concentrations(P 0.05).Dietary supplementation with AA significantly decreased GLO enzyme activity but increased splenic AA concentration and anti-bovine serum albumin IgG levels(P 0.05) and tended to increase CD4+T lymphocyte numbers(P = 0.06) in serum.Supplementation of 0.25 g AA/kg diet significantly increased hepatic total antioxidant capacity(T-AOC,P 0.05) relative to the control group.Conclusions: Laying hens could synthesize AA in both the kidney and the liver,though the GLO enzyme activities were 100 times greater in kidneys than in livers.The old laying hens had greater absorption and reabsorption capacity and higher AA retention in some tissues that did the young hens.Dietary supplementation of AA can improve the health of old layers by enhancing immunity and antioxidant capacity.     

Estradiol inhibits hepatic stellate cell area and collagen synthesis in the chicken liver

Hepatic stellate cells (HSCs) are the main collagen‐producing cells in the liver. The HSC area and amount of collagen fibers are different between male and female chickens. This study was performed to confirm the effect of estradiol on collagen synthesis in the growing chicken liver. Blood estradiol levels in chicks were compared at 4 and 8 weeks of age, and the collagen fibril network in liver tissue was observed at 8 weeks by scanning electron microscopy. Intraperitoneal administrations of estradiol and tamoxifen to male and female chicks, respectively, were performed daily from 5 to 8 weeks of age. The areas of HSCs and collagen contents were measured in the liver tissue. The blood estradiol level was higher in females than in males, and the collagen fibril network was denser in males than in females at 8 weeks of age. Estradiol administration in males induced decreases in the HSC area and collagen content of the liver. Conversely, tamoxifen administration in females induced an increase in the HSC area but did not facilitate collagen synthesis. Based on these results, estradiol inhibits the area and collagen synthesis of HSCs in the growing chicken liver under normal physiological conditions.     

The effect of biological isolation and a molt-inducing regimen on the recovery of Mycoplasma gallisepticum from commercial Leghorn hens

Two trials were conducted to determine the effect of induced molt on the reisolation of Mycoplasma gallisepticum (MG) from commercial leghorn hens that had been eyedrop-inoculated with MG at 10 weeks of age. Chickens were maintained in a conventional floored chicken house on dry litter through 100 weeks of age. At age 64 weeks, 4 days (Trial 1), and at 100 weeks (Trial 2), hens were swabbed and cultured for MG and then molted in biological isolation units. Swabs were again taken at the end of each molt. No difference was observed in the number of MG isolations between molted hens and controls that did not undergo molting. However, a significant decrease in MG isolations was observed in both trials from swabs obtained when hens were housed on dry litter floors as compared with swabs taken from the same hens after 18 days (Trial 1) or 21 days (Trial 2) of confinement in isolation units.     

Gustducin is expressed in the taste buds of the chicken

We investigated the expression of gustducin in chicken taste buds using molecular biological, biochemical and immunohistochemical techniques. Expression of a gustducin‐like sequence was detected by RT‐PCR in the tissues containing taste buds, and corresponded to the predicted gustducin gene in the chicken. Expression of this sequence was not detected in the brain, heart, liver, pancreas, intestine, kidney and testis of the chicken. The expressed sequence had a high specificity for oral tissues that contained taste buds. These results suggest that the detected sequence was the chicken gustducin gene. Next, we generated a polyclonal antiserum against the chicken gustducin protein to observe its localization in the oral tissues. The results revealed that the chicken gustducin was specifically expressed in the taste buds. It is suggested that the chicken has a gustatory system mediated by gustducin, and chicken gustducin is a reliable marker for taste buds or taste cells. This is the first molecular biological, biochemical and immunohistochemical demonstration of the presence of gustducin in the chicken.     

Influence of F strain Mycoplasma gallisepticum infection on response of commercial layers to heat exposure

Commercial layers were inoculated with F strain Mycoplasma gallisepticum (MG) and housed in either conventional chicken houses or the lower-stress environment of biological isolation units. At the end of 2 weeks, all treatment groups were placed in environmental chambers and subjected to 4 hr of heat stress (40 C with a dew point of 21 C). Rectal temperature, an indicator of response to high heat, was monitored. Rectal temperatures of F strain MG-inoculated hens housed in the conventional chicken house environment were significantly higher than those of uninoculated controls, whereas rectal temperatures of hens held in isolation units were comparable to those of their uninoculated controls.     

二氢吡啶(diludin)抗蛋鸡脂肪肝的机理

30周龄罗曼蛋鸡 80 0只 ,随机分为 2组 (每组设 4个重复 ,每个重复 10 0只 ) ,分别饲喂添加二氢吡啶 (diludin) 0、15 0 m g/ kg的玉米豆粕日粮 ,分别于试验 2 0周和 40周采血和屠宰 ,研究了二氢吡啶抗蛋鸡脂肪肝的机制。结果显示 ,日粮中添加二氢吡啶 ,试验鸡于 5 0、70周龄时 ,肝脂率和腹脂率均显著降低 (P<0 .0 5或 P<0 .0 1) ;腹脂激素敏感脂酶 (HSL )活性显著提高 (P<0 .0 5或 P<0 .0 1) ,脂蛋白脂酶 (L PL )活性显著降低 (P<0 .0 5或 P<0 .0 1) ,肝脏中NADPH生成酶 - MDH活性显著降低 (P<0 .0 1) ;血清脂肪酶活性和游离脂肪酸、极低密度脂蛋白胆固醇含量显著提高 (P<0 .0 5或 P<0 .0 1) ;血清甘油三酯含量降低 (P<0 .0 5或 P<0 .0 1) ;血清超氧化物歧化酶和 T3 显著提高 (P<0 .0 5或 P<0 .0 1) ;肝脏、腹脂中环腺苷酸 (c AMP)含量显著提高 (P<0 .0 5 )。上述结果表明 ,二氢吡啶通过促进产蛋中后期蛋鸡肝脏中脂肪的迁移和体脂的分解 ,达到抗脂肪肝效果。     

Expression and localization of estrogen receptors α and β mRNA in medullary bone of laying hens

The aim of this study was to observe the expression and localization of estrogen receptor (ER) α and ER β mRNA in the medullary bone of laying hens. First, medullary bone, liver, kidney, and shell gland of the oviduct tissues were dissected from laying hens. Then, the total cellular RNA was isolated from each tissue specimen, and the ER α and ER β mRNA expression was observed using semiquantitative RT‐PCR. Second, the localization of ER α mRNA in the medullary bone was detected with in situ hybridization using digoxigenin‐11‐UTP‐labeled cRNA probes. As a result, the expression of ER α mRNA was higher than that of ER β mRNA in the medullary bone, liver, and shell gland of the oviduct from laying hens. In the kidney, ER α mRNA expression was lower than that of ER β mRNA. The expression pattern of ER α and ER β mRNA of the medullary bone was similar to that of the shell gland of the oviduct. Moreover, ER α mRNA was intensively expressed in osteoblasts on the medullary bone surface and bone marrow stromal cells but was not expressed in osteoclasts. These results suggest that in medullary bone, estrogen action may be regulated not by ER β but by ER α.     

Serum-free culture of adult chicken hepatocytes; morphological and biochemical characterisation

The morphological and biochemical characterisation of adult chicken hepatocytes in a serum-free culture are described. When cultured in positively charged plastic dishes, chicken hepatocytes formed a monolayer cell sheet. The monolayer morphology of these chicken hepatocytes was quite distinct from the spheroid shape of rat hepatocytes cultured under similar conditions. Electron microscopy showed that the cytoplasmic organelles of chicken hepatocytes were well preserved in vitro. Two-dimensional gel electrophoresis showed that the chicken hepatocytes secreted liver-specific proteins. Several enzymes of glucose-6-phosphatase, cytochrome P-450 or glutathione S-transferase, involved in metabolic and biotransformation pathways in the liver, were retained in the chicken hepatocytes in a serum free condition. These findings suggest that the primary culture of adult chicken hepatocytes with a serum-free culture system could be useful to study the hepatic metabolic pathway in the chicken and its response to various chemicals.     

不同生理阶段边鸡卵巢转录谱的构建及卵泡发育相关基因的分析

本研究通过分析不同生理阶段边鸡卵巢转录表达谱,旨在找出与卵泡发育相关的基因并进行验证,以揭示边鸡就巢性强的遗传机制.构建代表不同生理期即70日龄(70 d,卵巢发育期)、165日龄(165 d,开产期)、220日龄(220 d,产蛋高峰期)和330日龄(330 d,就巢期)的边鸡卵巢基因表达谱,用转录组表达定量(RSE...