1、[中文题目]

          卵巢中B4GALNT2基因的异位表达与拉卡恩羊高繁殖力的关系研究

[全部的英文单位]     

  INRA-ENVT, UMR 444 ,Laboratoire de Génétique Cellulaire, Castanet- Tolosan, France.

[全部的中文单位]

法国农业科学研究院图卢兹国立兽医学校，UMR 444，细胞遗传学实验室，卡斯塔内托洛桑，法国。

[出处]

     Drouilhet L; Mansanet C; Sarry J; Tabet K; Bardou P; Woloszyn F; Lluch J; Harichaux G;Viguie C; Monniaux D; Bodin L; Mulsant P; Fabre S. The Highly Prolific Phenotype of Lacaune Sheep Is Associated with an Ectopic Expression of the B4GALNT2 Gene within the Ovary. PLOS GENETICS, 2013, 9 (9): e 1003809.

[ABSTRACT]

  Prolific sheep have proven to be a valuable model to identify genes and mutations implicated in female fertility. In the Lacaune sheep breed, large variation in litter size is genetically determined by the segregation of a fecundity major gene influencing ovulation rate, named FecL and its prolific allele FecL(L). Our previous work localized FecL on sheep chromosome 11 within a locus of 1.1 Mb encompassing 20 genes. With the aim to identify the FecL gene, we developed a high throughput sequencing strategy of long-range PCR fragments spanning the locus of FecL(L) carrier and non-carrier ewes. Resulting informative markers defined a new 194.6 kb minimal interval. The reduced FecL locus contained only two genes, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) and beta-1,4-N-acetyl-galactosaminyl transferase 2 (B4GALNT2), and we identified two SNP in complete linkage disequilibrium with FecL(L). B4GALNT2 appeared as the best positional and expressional candidate for FecL, since it showed an ectopic expression in the ovarian follicles of FecL(L)/FecL(L) ewes at mRNA and protein levels. In FecL(L) carrier ewes only, B4GALNT2 transferase activity was localized in granulosa cells and specifically glycosylated proteins were detected in granulosa cell extracts and follicular fluids. The identification of these glycoproteins by mass spectrometry revealed at least 10 proteins, including inhibin alpha and betaA subunits, as potential targets of B4GALNT2 activity. Specific ovarian protein glycosylation by B4GALNT2 is proposed as a new mechanism of ovulation rate regulation in sheep, and could contribute to open new fields of investigation to understand female infertility pathogenesis.

[中文摘要]

 人们已经证实产仔数高的羊是一个很有价值的模型，可以用来识别与母羊繁殖能力相关的基因和突变。从基因方面来讲，拉卡恩羊窝产仔数的不同是由与繁殖力相关的FecL基因和它的等位基因FecL(L)之间分离决定的，FecL是影响排卵率的主效基因。我们之前的工作是在绵羊11号染色体包含20个基因的1.1Mb位点处定位FecL基因。为了检测FecL基因，我们自主开发了一个长片段PCR技术和高通量测序分析，用来衡量母羊中是否有携带FecL(L)基因的位点。一些已知信息的标记界定了一个全新的194.6kb最小片段间隔。缩减的FecL位点只包含两个基因，胰岛素样生长因子ⅡmRNA结合蛋白(IGF2BP1)和β1,4-N-乙酰氨基半乳糖转移酶2(B4GALNT2)。FecL(L)基因存在完全连锁不平衡，我们在其中检测到两个单核苷酸多态性。无论在位置上还是表达上，B4GALNT2都是作为FecL的最佳候选基因，因为它能在mRNA和蛋白水平上表明FecL(L)/FecL(L)母羊卵泡的异位表达。对于只有FecL(L)的母羊，B4GALNT2转移酶活性作用于颗粒细胞，特别是在颗粒细胞提取物和卵泡液中发现糖基化蛋白。用质谱分析法检测这些糖蛋白发现至少有十种蛋白质，包括抑制素α和βA亚基，是B4GALNT2作用的潜在靶区域。因此认为由B4GALNT2作用导致特定的卵巢蛋白糖基化是调控绵羊排卵率的全新机制，并且有利于打开新的思路和方法来了解雌性不孕的发病机理。

2、[中文题目]

          全基因组关联分析鉴定绵羊的异常高产与BMP15基因的两处新突变相关

[全部的英文单位]

  INRA, UMR 444 Laboratoire de Génétique Cellulaire, Castanet-Tolosan, France.

[全部的中文单位]

  法国农业科学研究院，UMR 444，细胞遗传学实验室，卡斯塔内托洛桑，法国。

[出处]

  Demars J, Fabre S, Sarry J, Rossetti R, Gilbert H, Persani L, Tosser-Klopp G, Mulsant P, Nowak Z, Drobik W, Martyniuk E, Bodin L. Genome-wide association studies identify two novel BMP15 mutations responsible for an atypical hyperprolificacy phenotype in sheep. PLOS GENETICS, 2013, Apr, 9(4): e1003482.

[ABSTRACT]  

  Some sheep breeds are naturally prolific, and they are very informative for the studies of reproductive genetics and physiology. Major genes increasing litter size (LS) and ovulation rate (OR) were suspected in the French Grivette and the Polish Olkuska sheep populations, respectively. To identify genetic variants responsible for the highly prolific phenotype in these two breeds, genome-wide association studies (GWAS) followed by complementary genetic and functional analyses were performed. Highly prolific ewes (cases) and normal prolific ewes (controls) from each breed were genotyped using the Illumina Ovine SNP50 Genotyping Beadchip. In both populations, an X chromosome region, close to the BMP15 gene, harbored clusters of markers with suggestive evidence of association at significance levels between 1E(-05) and 1E(-07). The BMP15 candidate gene was then sequenced, and two novel non-conservative mutations called FecX(Gr) and FecX(O) were identified in the Grivette and Olkuska breeds, respectively. The two mutations were associated with the highly prolific phenotype (p FecX (Gr) = 5.98E(-06) and p FecX(O) = 2.55E(-08)). Homozygous ewes for the mutated allele showed a significantly increased prolificacy (FecX(Gr)/FecX(Gr), LS = 2.50 ± 0.65 versus FecX(+)/FecX(Gr), LS = 1.93 ± 0.42, p<1E(-03) and FecX(O)/FecX(O), OR = 3.28 ± 0.85 versus FecX(+)/FecX(O), OR = 2.02 ± 0.47, p<1E(-03)). Both mutations are located in very well conserved motifs of the protein and altered the BMP15 signaling activity in vitro using a BMP-responsive luciferase test in COV434 granulosa cells. Thus, we have identified two novel mutations in the BMP15 gene associated with increased LS and OR. Notably, homozygous FecX(Gr)/FecX(Gr) Grivette and homozygous FecX(O)/FecX(O) Olkuska ewes are hyperprolific in striking contrast with the sterility exhibited by all other known homozygous BMP15 mutations. Our results bring new insights into the key role played by the BMP15 protein in ovarian function and could contribute to a better understanding of the pathogenesis of women's fertility disorders.

[中文摘要]  

  一些绵羊品种天生具有高繁殖力，并且能够用于研究繁殖遗传学和生理学。在法国Grivette羊和波兰的Olkuska羊中分别检测到可能提高产仔数和排卵率的主效基因。为了确认在这两个品种中遗传突变对超高繁殖力有影响，本实验对补充的基因和功能分析之后进行全基因组关联分析。使用Illumina绵羊SNP50基因分型芯片，分别在每个品种中将超高产的母羊（病例组）和正常高产的母羊（对照组）进行基因分型。在这两个群体中，X染色体上靠近BMP15基因的区域存在一些标记，这些标记在显著水平上[1E(-05) ~1E(-07)]证明与高繁殖力有关系。之后对BMP候选基因进行测序，分别在Grivette和Olkuska羊品种中检测到两处新的非保守突变，分别被称之为FecX(Gr) 和 FecX(O)。这两个突变和超高产相关(p FecX (Gr) = 5.98E(-06) ， p FecX(O) = 2.55E(-08))。等位基因发生突变的纯合子母羊的繁殖力显著地增加(FecX(Gr)/FecX(Gr), LS = 2.50 ± 0.65 versus FecX(+)/FecX(Gr), LS = 1.93 ± 0.42, p<1E(-03) and FecX(O)/FecX(O), OR = 3.28 ± 0.85 versus FecX(+)/FecX(O), OR = 2.02 ± 0.47, p<1E(-03))。这两个突变都位于蛋白质非常保守的基序上，并且使用BMP应答的荧光素酶检测COV434颗粒细胞发现它们改变了BMP15在体外的信号强弱。因此，我们认为BMP15基因的两处新突变和LS以及OR的升高有关联。尤其是，相比于其他已知的纯合子BMP15突变导致的不孕不育，纯合子FecX(Gr)/FecX(Gr) Grivette母羊以及FecX(O)/FecX(O) Olkuska母羊的繁殖力显著地高。本实验结果引进了关于BMP15蛋白在卵巢功能上发挥关键作用的全新认识，并且有助于更好地理解女性生殖疾病的发病机理。

3、[中文题目]

       研究简报：Latxa奶绵羊的朊病毒蛋白(PRNP)基因位点和泌乳性状之间的关联性分析

[全部的英文单位]

  INRA, UMR 1289, Tissus Animaux Nutrition Digestion Ecosystème et Métabolisme, F-31326 Castanet-Tolosan, France.

[全部的中文单位]

  法国农业科学研究院， UMR 1289，动物组织营养消化生态系统和新陈代谢，F-31326卡斯塔内托洛桑，法国。

[出处]

   Vitezica ZG, Beltran de Heredia I, Ugarte E. Short communication: Analysis of association between the prion protein (PRNP) locus and milk traits in Latxa dairy sheep. J Dairy Sci. 2013 Sep;96(9):6079-83.

[ABSTRACT]

    The purpose of this study was to analyze associations between polymorphisms in the PRNP gene and ewe milk traits. A total of 242,565 lactations of the Latxa breed were used. Milk, fat and protein yields, and fat and protein content from black-faced Latxa from Spanish Basque Country, black-faced Latxa from Navarra, and blond-faced Latxa were collected. To evaluate evidence of association, the different traits were analyzed using an animal model, where the PRNP genotype effect was included or not as a random effect. Adding the PRNP effect to the model improved the fitting for milk yield in black-faced Latxa from Spanish Basque Country and in blond-faced Latxa, for fat yield in black-faced Latxa from Navarra, and for protein yield in blond-faced Latxa. However, the proportion of the phenotypic variance explained by the PRNP effect for milk yield (1.0×10(-3)), fat yield (3.6×10(-3)) and protein yield (9.4×10(-4)) were near zero. The PRNP locus accounts for about 0.5, 1.5, and 0.4% of total genetic (PRNP and polygenic) variance in milk, fat, and protein yield. These values indicated that the PRNP effect is not relevant regarding genetic additive contribution. For breeding purposes, it is unlikely that selection for scrapie resistance will have an effect on the milk traits studied in the Latxa breed.

[中文摘要]

 本研究的目的是分析PRNP基因的多态性与母羊泌乳性状的相互关系。采用Latxa 泌乳期的绵羊共242,565只，包括西班牙巴斯克地区的黑面Latxa品种、纳瓦拉地区的黑面Latxa品种和棕面Latxa品种。收集它们的泌乳量、脂肪含量和蛋白含量这些数据。为了证明PRNP基因的多态性与母羊泌乳性状的关联性，使用同一个动物模型进行不同性状的分析，其中有PRNP基因型效应作用或者只是随机效应。提高模型动物内的PRNP基于效应能够提高西班牙巴斯克黑面Latxa羊的泌乳量，提高纳瓦拉黑面Latxa羊的乳脂量以及提高棕面Latxa羊的乳蛋白含量。然而，PRNP基因效应对泌乳量(1.0×10(-3))、乳脂量(3.6×10(-3))及乳蛋白量(9.4×10(-4)) 表型方差百分比接近0。PRNP基因位点在泌乳量、乳脂量和乳蛋白量中全部基因（PRNP和多基因的）变化所占的比例大约0.5，1.5和0.4%。这些数值表明，PRNP效应和基因加性效应不相关。对于育种目标来说，选择Latxa品种抗痒病羊不见得对泌乳性状有影响。

4、[中文题目]

          妊娠早期绵羊乳腺的MicroRNA：miR-21, miR-205, and miR-200的时空表达

[全部的英文单位]

 Institut National de la Recherche Agronomique (INRA), Unité de Recherche 1196. Génomique et Physiologie de la Lactation, Jouy-en-Josas, France.

[全部的中文单位]

    法国农业科学研究院，联合研究中心1196，泌乳期基因组学和生理学，茹依桑若萨，法国。

[出处]

   Galio L, Droineau S, Yeboah P, Boudiaf H, Bouet S, Truchet S, Devinoy E.MicroRNA in the ovine mammary gland during early pregnancy: ，spatial and temporal expression of miR-21, miR-205, and miR-200. Physiol Genomics. 2013 Feb 15;45(4):151-61.

[ABSTRACT]

    The mammary gland undergoes extensive remodeling between the beginning of pregnancy and lactation; this involves cellular processes including cell proliferation, differentiation, and apoptosis, all of which are under the control of numerous regulators. To unravel the role played by miRNA, we describe here 47 new ovine miRNA cloned from mammary gland in early pregnancy displaying strong similarities with those already identified in the cow, human, or mouse. A microarray study of miRNA variations in the adult ovine mammary gland during pregnancy and lactation showed that 100 miRNA are regulated according to three principal patterns of expression: a decrease in early pregnancy, a peak at midpregnancy, or an increase throughout late pregnancy and lactation. One miRNA displaying each pattern (miR-21, miR-205, and miR-200b) was analyzed by qRT-PCR. Variations in expression were confirmed for all three miRNA. Using in situ hybridization, we detected both miR-21 and miR-200 in luminal mammary epithelial cells when expressed, whereas miR-205 was expressed in basal cells during the first half of pregnancy and then in luminal cells during the second half. We therefore conclude that miR-21 is strongly expressed in the luminal cells of the normal mammary gland during early pregnancy when extensive cell proliferation occurs. In addition, we show that miR-205 and miR-200 are coexpressed in luminal cells, but only during the second half of pregnancy. These two miRNA may cooperate to maintain epithelial status by repressing an EMT-like program, to achieve and preserve the secretory phenotype of mammary epithelial cells.

[中文摘要]

  在怀孕初期到开始泌乳期间，乳腺经历了一个伸张重塑的过程。它涉及到细胞进化，包括细胞增殖、分化以及凋亡，都受到许多调控因子的作用。为了阐明microRNA的作用机制，这里我们对怀孕初期的绵羊克隆了47个miRNA，证实了其在牛、人和老鼠中高度的相似。通过基因芯片技术研究在妊娠期和泌乳期的成年母羊乳腺的miRNA变化状态，发现100种miRNA被调控后主要有三种表达形式：妊娠初期降低，妊娠中期最高，或者在整个妊娠后期和泌乳期升高。qRT-PCR分析每个miRNA显现地形式(miR-21, miR-205, and miR-200b)。这三个miRNA已被证实表达存在变化。运用原位分子杂交，我们检测到miR-21和miR-200在乳腺上皮细胞的细胞腔表达，然而妊娠初期miR-205在基底细胞中表达，妊娠后期在细胞腔中表达。因此，我们得到结论，在妊娠初期大量的细胞增殖时，正常乳腺的细胞腔中有miR-200表达。另外，我们发现miR-205和miR-200在细胞腔中共同表达，但是只是在妊娠后期。通过抑制类似于上皮间质转化的机制，这两种miRNA可能协同维持上皮细胞状态来获得或保护乳腺上皮细胞的具有分泌作用的基因型。

5、[中文题目]

          光周期变化中的下丘脑神经细胞粘附分子的唾液酸形态与绵羊产羔数相关关系

[全部的英文单位]

   INRA, UMR85 Physiologie de la Reproduction et des Comportements, Nouzilly, France.

[全部的中文单位]

  法国农业科学研究院，联合研究中心85，繁殖生理学和行为学，努齐利，法国。

[出处]

  Chalivoix S, Guillaume D, Cognié J, Thiéry JC, Malpaux B, Dufourny L. Photoperiodic variations of the polysialylated form of neural cell adhesion molecule within the hypothalamus and related reproductive output in the ewe.Cell Tissue Res. 2013 May;352(2):387-99.

[ABSTRACT]

   Cellular mechanisms induced by melatonin to synchronise seasonal reproduction in several species, including sheep, remain unclear. We sought to evaluate the scale and physiological significance of neural plasticity in order to explain the delay between the change of duration of melatonin secretion and the change of reproductive status following a transition from long days (LD, 16 h light/24 h) to short days (SD, 8 h light/24 h) and from SD to LD. Using Western blots in ovariectomised oestradiol-replaced ewes, we evaluated the content of the polysialylated form of neural cell adhesion molecule (PSA-NCAM), a plasticity marker, in the hypothalamus. From day 15 following a transition to SD, most hypothalamic areas showed a decrease of PSA-NCAM level that was particularly significant in the preoptic area (POA). Following a transition to LD, PSA-NCAM content increased at day 15 in most regions except in the premammillary hypothalamic area (PMH) in which a significant decrease was noted. The functional importance of PSA-NCAM variations for seasonal reproduction was assessed for the PMH and POA. PSA-NCAM was degraded by stereotaxic injections of endoneuraminidase N and luteinising hormone (LH) secretion was recorded in treated and control ewes. Degradation of PSA-NCAM in the PMH in SD-treated ewes failed to produce a significant effect on LH secretion, whereas a similar treatment in the POA before a transition to SD delayed activation of the gonadotroph axis in two-thirds of the ewes. Our results suggest that the photoperiod controls variations of the hypothalamic content of a plasticity marker and that these might be important for the regulation of seasonal reproduction, particularly in the POA.

[中文摘要]

  褪黑激素诱导细胞机制来同步数个物种间包括绵羊的季节性繁殖，这一观点尚不可知。我们试图评估神经可塑性的大小和生理意义，是为了解释从长日照（LD 16 h 光照/24 h）到短日照（SD 8 h 光照/24 h）以及短日照到长日照转变时褪黑激素分泌的持续时间和生殖状态的变化。对母羊卵巢上的雌二醇进行免疫印迹，我们评估的内容是下丘脑中神经细胞黏附分子唾液酸（PSA - NCAM）的形态，它是一种可塑性标记。从SD过渡后的第15天，下丘脑PSA-NCAM水平降低，尤其是在视前区（POA）有极显著下降。过渡到LD的第15天， PSA-NCAM含量除了在下丘脑乳头突起（PMH）有显著降低，在大多数区域都增加。通过PMH和POA来评估PSA-NCAM变化对季节性繁殖的功能性重要作用。采用立体定向注射内N-神经氨酸苷酶后PSA-NCAM水平降低，然后记录处理组和对照组母羊的促黄体激素（LH）的分泌。对于SD-处理组母羊，PMH部位PSA- NCAM水平降低不会对LH分泌产生显著作用，而相似的处理组——转换为SD之前，在POA区域，PSA- NCAM延迟激活三分之二母羊的促性腺轴。我们的结论表明，光周期控制下丘脑，尤其是在视前区中的可塑性标记的变化，这些可能会对季节性繁殖的调控具有重要作用。

相关信息:

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