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Ruminant reproduction: recent findings and future challenges, a summary. 反刍动物繁殖:最近的发现和未来的挑战,总结。
E K Inskeep
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引用次数: 0
Assisted reproduction in Mediterranean wild ruminants: lessons from the Spanish ibex (Capra pyrenaica). 地中海野生反刍动物的辅助繁殖:来自西班牙野山羊的经验。
Pub Date : 2010-01-01 DOI: 10.5661/RDR-VII-431
J. Santiago-Moreno, M. A. Coloma, A. Toledano-Díaz, C. Castaño, A. Gómez-Brunet, A. López-Sebastián
Despite apparent progress in reproductive technology as applied to wild ruminants, the success achieved in terms of the number of offspring that become healthy adults has remained low. Difficulties often arise through a lack of knowledge regarding appropriate cryopreservation techniques, and indeed through a lack of detailed information on the reproductive physiology of the species in question. The Spanish ibex (Capra pyrenaica) is a wild caprid found exclusively in the mountains of Iberia; only two of the original four subspecies still exist. Great efforts need to be made to preserve this species. The endocrine and environmental mechanisms that control its seasonal reproduction need to be properly understood, reproductive technologies (particularly the cryopreservation of gametes) optimised, and genetic resource banks developed. The experience obtained with the Spanish ibex may be useful in ex situ conservation strategies designed to preserve other threatened Mediterranean wild ruminants.
尽管应用于野生反刍动物的生殖技术取得了明显进展,但在成为健康成年动物的后代数量方面取得的成功仍然很低。困难往往是由于缺乏关于适当的低温保存技术的知识,实际上是由于缺乏有关物种生殖生理学的详细信息。西班牙野山羊(Capra pyrenaica)是一种只在伊比利亚山脉发现的野生野山羊;最初的四个亚种中只有两个仍然存在。需要做出巨大的努力来保护这个物种。需要正确理解控制其季节性繁殖的内分泌和环境机制,优化生殖技术(特别是配子的低温保存),并开发遗传资源库。从西班牙野山羊身上获得的经验可能对旨在保护其他受威胁的地中海野生反刍动物的迁地保护策略有用。
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引用次数: 7
Genomic tools for characterizing monogenic and polygenic traits in ruminants--using the bovine as an example. 描述反刍动物单基因和多基因特征的基因组工具——以牛为例。
J F Taylor, R H Chapple, J E Decker, S J Gregg, J W Kim, S D McKay, H R Ramey, M M Rolf, T M Taxis, R D Schnabel

Next generation sequencing platforms have democratized genome sequencing. Large genome centers are no longer required to produce genome sequences costing millions. A few lanes of paired-end sequence on an Illumina Genome Analyzer, costing < $10,000, will produce more sequence than generated only a few years ago to produce the human and cow assemblies. The de novo assembly of large numbers of short reads into a high-quality whole-genome sequence is now technically feasible and will allow the whole genome sequencing and assembly of a broad spectrum of ruminant species. Next-generation sequencing instruments are also proving very useful for transcriptome or resequencing projects in which the entire RNA population produced by a tissue, or the entire genomes of individual animals are sequenced, and the produced reads are aligned to a reference assembly. We have used this strategy to examine gene expression differences in tissues from cattle differing in feed efficiency, to perform genome-wide single nucleotide polymorphism discovery for the construction of ultrahigh-density genotyping assays, and in combination with genome-wide association analysis, for the identification of mutations responsible for Mendelian diseases. The new 800K SNP bovine genotyping assays possess the resolution to map trait associations to the locations of individual genes and the 45 million polymorphisms identified in > 180X genome sequence coverage on over 200 animals can be queried to identify the polymorphisms present within positional candidate genes. These new tools should rapidly allow the identification of genes and mutations underlying variation in cattle production and reproductive traits.

下一代测序平台使基因组测序大众化。大型基因组中心不再需要花费数百万美元来制作基因组序列。在Illumina基因组分析仪上,花费不到1万美元的几列成对末端序列,将产生比几年前产生的人类和奶牛组装更多的序列。将大量短序列重新组装成高质量的全基因组序列现在在技术上是可行的,并将允许对广泛的反刍动物物种进行全基因组测序和组装。新一代测序仪器也被证明对转录组或重测序项目非常有用,在这些项目中,对组织产生的整个RNA群体或单个动物的整个基因组进行测序,并将产生的reads与参考汇编对齐。我们使用这种策略来检测不同饲料效率的牛组织中的基因表达差异,进行全基因组单核苷酸多态性发现,以构建超高密度基因分型分析,并结合全基因组关联分析,以确定导致孟德尔疾病的突变。新的800K SNP牛基因分型分析具有将性状关联定位到单个基因位置的分辨率,并且可以查询超过200只动物的> 180X基因组序列覆盖范围内鉴定的4500万个多态性,以确定定位候选基因内存在的多态性。这些新工具应该能够迅速识别牛生产和繁殖性状变异的基因和突变。
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引用次数: 0
Inter- and intra-cellular mechanisms of prostaglandin F2alpha action during corpus luteum regression in cattle. 牛黄体退化过程中前列腺素F2alpha作用的细胞间和细胞内机制。
Pub Date : 2010-01-01 DOI: 10.7313/upo9781907284991.025
D J Skarzynski, K Okuda

The bovine corpus luteum (CL) grows very fast and regresses within a few days at luteolysis. Mechanisms controlling development and secretory function of the bovine CL may involve many factors that are produced both within and outside the CL. In the cow, luteolysis is initiated by uterine prostaglandin (PG)F2alpha released at the late luteal stage. It can also be induced by injection of exogenous PGF2alpha given at the mid luteal stage. Luteolysis consists of a phase of rapid decrease in progesterone (P4) production by the CL, followed by a phase of structural regression. Although uterine PGF2alpha is known to be the main luteolytic factor, its direct action on the CL is mediated by the products of accessory luteal cells: immune cells, endothelial cells, pericytes and fibroblasts. There are studies showing that beside endothelin-1, cytokines (tumor necrosis factor-alpha, interferons) and nitric oxide play critical roles in functional and structural luteolysis in cattle by stimulating leukotrienes and PGF2alpha', decreasing P4 secretion and apoptosis induction. Because of luteal blood flow and P4 concentrations decrease in parallel during both spontaneous and PGF2alpha-induced luteolysis, a decrease in luteal blood flow resulting in hypoxia has been proposed as one of the main luteolytic mechanisms in the cow. Hypoxia inhibits P4 synthesis in luteal cells by inhibiting the steroidogenic enzymes and promotes apoptosis of luteal cells by increasing pro-apoptotic proteins. Although reduction of luteal blood flow and hypoxia contribute to the late events of luteolysis, little is known about the physiological relevance and the cause of the transient increase in luteal blood flow and reactive oxygen species during the initial step of luteolysis.

牛黄体(CL)生长非常快,并在几天内退行黄体溶解。控制牛乳糜泻发育和分泌功能的机制可能涉及乳糜泻内部和外部产生的许多因素。在奶牛中,黄体溶解是由黄体晚期释放的子宫前列腺素(PG)F2alpha引起的。在黄体中期注射外源性PGF2alpha也可诱导。黄体溶解包括一个由CL快速减少黄体酮(P4)产生的阶段,随后是一个结构回归的阶段。虽然已知子宫PGF2alpha是主要的黄体溶解因子,但其对CL的直接作用是由黄体附属细胞的产物介导的:免疫细胞、内皮细胞、周细胞和成纤维细胞。有研究表明,除内皮素-1外,细胞因子(肿瘤坏死因子- α、干扰素)和一氧化氮通过刺激白三烯和pgf2 α′,减少P4分泌和诱导细胞凋亡,在牛的功能性和结构性黄体溶解中起关键作用。由于黄体血流量和P4浓度在自发和pgf2α诱导的黄体溶解过程中平行减少,黄体血流量减少导致缺氧被认为是牛黄体溶解的主要机制之一。缺氧通过抑制甾体生成酶抑制黄体细胞P4合成,并通过增加促凋亡蛋白促进黄体细胞凋亡。虽然黄体血流量的减少和缺氧有助于黄体溶解的后期事件,但在黄体溶解的初始阶段,黄体血流量和活性氧的短暂增加的生理相关性和原因尚不清楚。
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引用次数: 19
Managing the dominant follicle in high-producing dairy cows. 高产奶牛的显性卵泡管理。
Pub Date : 2010-01-01 DOI: 10.7313/upo9781907284991.020
M C Wiltbank, R Sartori, J L M Vasconcelos, A B Nascimento, A H Souza, A P Cunha, A Gumen, S Sangsritavong, J N Guenther, H Lopez, J R Pursley

Reduced reproductive efficiency has been reported in high-producing dairy cows. Sources of reproductive inefficiency include decreased expression of estrus, increased diameter of the ovulatory follicle and reduced fertility when cows are inseminated after estrus, increased incidence of double ovulation and twinning, and increased pregnancy loss. To overcome some of these inefficiencies, reproductive management programs have been developed that synchronize ovulation and enable effective timed artificial insemination (AI) of lactating dairy cows. Effective regulation of the corpus luteum (CL), follicles, and hormonal environment are critical for optimizing these programs. Recent programs, such as the 5-day CIDR program, Double-Ovsynch, G-6-G, and estradiol benzoate-CIDR programs were designed to more effectively control one or more physiological events. These events include synchronization of a new follicular wave at the beginning of the program, optimization of the circulating progesterone (P4) concentrations and duration of follicular dominance, optimized reductions in P4 and increases in circulating estradiol (E2) concentrations during the preovulatory period, and tightly synchronized ovulation of a follicle of optimal size and fertility for implementation of timed AI. The success of these programs has been remarkable, although there is substantial variability in effectiveness due to environmental, management, nutritional, genetic, and disease factors as well as potential variability in some aspects of reproductive physiology among commercial dairy farms. Future programs will optimize the reproductive physiology while simplifying the protocol implementation and also match specific reproductive management protocols to specific farms and even specific cows (for example primiparous vs. multiparous).

据报道,高产奶牛的繁殖效率降低。生殖效率低下的原因包括发情期表达降低、卵泡直径增大和发情期后受精卵受精率降低、双排卵和双胞胎发生率增加以及妊娠损失增加。为了克服这些低效率的问题,人们开发了生殖管理程序,以同步排卵并实现有效的定时人工授精(AI)。黄体(CL)、卵泡和激素环境的有效调节是优化这些程序的关键。最近的方案,如5天CIDR方案,双ovsync, G-6-G和雌二醇-苯甲酸酯CIDR方案,旨在更有效地控制一个或多个生理事件。这些事件包括在程序开始时新卵泡波的同步,优化循环孕酮(P4)浓度和卵泡优势持续时间,优化排卵期P4的降低和循环雌二醇(E2)浓度的增加,以及为实施定时人工智能而实现最佳大小和生育能力的卵泡的紧密同步排卵。尽管由于环境、管理、营养、遗传和疾病因素以及商业奶牛场生殖生理某些方面的潜在差异,这些项目的有效性存在很大差异,但这些项目的成功是显著的。未来的项目将优化生殖生理学,同时简化协议的实施,并将特定的生殖管理协议与特定的农场甚至特定的奶牛(例如初产与多产)相匹配。
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引用次数: 6
Dietary regulation of developmental programming in ruminants: epigenetic modifications in the germline. 反刍动物发育程序的饮食调节:种系的表观遗传修饰。
Pub Date : 2010-01-01 DOI: 10.5661/RDR-VII-59
K. Sinclair, A. Karamitri, D. Gardner
Ruminants have been utilised extensively to investigate the developmental origins of health and disease, with the sheep serving as the model species of choice to complement dietary studies in the rat and mouse. Surprisingly few studies, however, have investigated delayed effects of maternal undernutrition during pregnancy on adult offspring health and a consistent phenotype, together with underlying mechanistic pathways, has not emerged. Nevertheless, when broad consideration is given to all studies with ruminants it is apparent that interventions that are initiated very early in gestation, and/or prior to conception, lead to greater effects on adult physiology than those that are specifically targeted to late gestation. Effects induced following dietary interventions at the earliest stages of mammalian development have been shown to arise as a consequence of alterations to key epigenetic processes that occur in germ cells and pluripotent embryonic cells. Currently, our understanding of epigenetic programming in the germline is greatest for the mouse, and is considered in detail in this article together with what is known in ruminants. This species imbalance, however, looks set to change as fully annotated genomic maps are developed for domesticated large animal species, and with the advent of 'next-generation' DNA sequencing technologies that have the power to globally map the epigenome at single-base-pair resolution. These developments would help to address such issues as sexually dimorphic epigenetic alterations to DNA methylation that have been found to arise following dietary restrictions during the peri-conceptional period, the effects of paternal nutritional status on epigenetic programming through the germline, and transgenerational studies where, in future, greater emphasis in domesticated ruminants should be placed on traits of agricultural importance.
反刍动物已被广泛用于研究健康和疾病的发育起源,绵羊可作为大鼠和小鼠饮食研究的补充模式物种。然而,令人惊讶的是,很少有研究调查了怀孕期间母亲营养不良对成年后代健康的延迟影响,并且没有出现一致的表型以及潜在的机制途径。然而,当对所有反刍动物的研究进行广泛考虑时,很明显,在妊娠早期和/或怀孕之前开始的干预措施对成年生理的影响比那些专门针对妊娠后期的干预措施更大。在哺乳动物发育的早期阶段,饮食干预引起的影响已被证明是生殖细胞和多能胚胎细胞中发生的关键表观遗传过程改变的结果。目前,我们对生殖系的表观遗传编程的了解最多的是小鼠,本文将与反刍动物的已知情况一起详细讨论。然而,随着驯化的大型动物物种的全注释基因组图谱的开发,以及“下一代”DNA测序技术的出现,这种物种不平衡似乎将会改变,这些技术具有以单碱基对分辨率绘制全球表观基因组图谱的能力。这些发展将有助于解决以下问题:在怀孕期饮食限制后发现的DNA甲基化的性别二态表观遗传改变,父亲营养状况对生殖系表观遗传编程的影响,以及跨代研究,在未来,驯化反刍动物应该更加重视农业重要性的性状。
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引用次数: 8
Spermatogonial stem cell biology in the bull: development of isolation, culture, and transplantation methodologies and their potential impacts on cattle production. 公牛精原干细胞生物学:分离、培养和移植方法的发展及其对牛生产的潜在影响。
J M Oatley

Widespread adoption of artificial insemination as a breeding practice has allowed for expanded use of desirable genetics from specific sires and greatly influenced production traits in dairy cattle populations worldwide. In fact, the average dairy cow in the US in 2009 produced 4.5 times more milk than in 1940 when commercialization of artificial insemination began. While many factors have contributed to this rapid increase in levels of milk production, genetic gain through expanded utilization of germlines from specific sires has been a major contribution. In comparison, use of artificial insemination in beef cattle populations has been limited due to challenges with implementing intensive management strategies required for success. Thus, there is need for alternative reproductive tools to expand use of desirable male genetics in the beef cattle industry. The process of sperm production, termed spermatogenesis, is supported by a tissue-specific stem cell population referred to as spermatogonial stem cells (SSCs). These unique cells have the capacity for infinite self-renewal and long-term regeneration of spermatogenesis following transplantation. In rodents, methods for isolating, culturing, and transplanting SSCs have been devised. For beef cattle, transplanting SSCs isolated from a donor male into the testes of recipient males in which donor-derived spermatogenesis occurs and offspring with donor genetics are produced from natural breeding has great potential as an alternative to artificial insemination. This potential reproductive strategy would allow for expansive use of genetics from desirable sires that overcomes the logistical challenges of artificial insemination. Translation of the methods devised for rodents to cattle is at the forefront of development. Devising means for isolating an SSC-enriched cell fraction from donor testes and identifying conditions that support long-term maintenance and proliferation of bovine SSCs in vitro are two tools that would greatly accelerate the pace at which transplantation will become a commercially viable option for cattle industries. Recent studies showed that expression of THY1 by SSCs is a conserved phenotype between rodents and cattle, and selection of the THY1 + fraction from donor testes can be used for isolating an SSC-enriched germ cell population. In addition, the conditions devised for expanding the number of rodent SSCs in vitro continues to serve as the basis for developing conditions that support bovine SSCs. With these tools in hand major advances in developing implementable reproductive tools with SSCs for commercial cattle production will be made in the coming decade.

人工授精作为一种育种方法的广泛采用,已经允许扩大使用特定品种的理想基因,并极大地影响了全世界奶牛种群的生产性状。事实上,2009年美国平均每头奶牛的产奶量是1940年人工授精商业化开始时的4.5倍。虽然许多因素促成了牛奶产量水平的快速增长,但通过扩大利用来自特定奶牛的种系而获得的遗传增益是一个主要贡献。相比之下,由于实施成功所需的集约化管理战略的挑战,在肉牛种群中人工授精的使用受到限制。因此,需要替代生殖工具,以扩大在肉牛工业中使用理想的雄性遗传学。精子产生的过程,称为精子发生,是由组织特异性干细胞群称为精原干细胞(ssc)支持的。这些独特的细胞在移植后具有无限自我更新和长期再生精子的能力。在啮齿类动物中,已经设计了分离、培养和移植ssc的方法。对于肉牛来说,将从供体雄性中分离的ssc移植到供体雄性的睾丸中,从而发生供体来源的精子发生,并通过自然育种产生具有供体遗传的后代,作为人工授精的替代方案具有很大的潜力。这种潜在的生殖策略将允许从理想的品种中广泛使用遗传学,从而克服人工授精的后勤挑战。将啮齿类动物的方法转化为牛的方法是发展的前沿。设计从供体睾丸中分离ssc富集细胞的方法和确定支持牛ssc在体外长期维持和增殖的条件是两种工具,这将大大加快移植成为养牛业商业上可行选择的步伐。最近的研究表明,ssc表达THY1在啮齿动物和牛之间是一种保守的表型,从供体睾丸中选择THY1 +片段可用于分离富含ssc的生殖细胞群体。此外,为扩大体外培养啮齿动物间充质干细胞数量而设计的条件继续作为开发支持牛间充质干细胞条件的基础。有了这些工具,未来十年将在开发可实施的带有ssc的用于商业牛生产的生殖工具方面取得重大进展。
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引用次数: 0
Developmental programming of the ovine placenta. 绵羊胎盘的发育程序。
A L Fowden, J W Ward, F B P Wooding, A J Forhead

The pattern of intrauterine growth and size at birth, in particular, programmes the structure and function of tissues later in life in many species, which has important implications for the incidence of adult-onset generative diseases in human populations. In mammals, the main determinant of intrauterine growth is the placental supply of nutrients which, in turn, depends on the size, morphology, transport characteristics and endocrine function of the placenta. However, compared to somatic tissues, little is known about the developmental programming of the placenta. This review examines the epigenetic regulation of placental phenotype with particular emphasis on the nutrient transfer capacity of the ovine placenta and environmental factors shown to cause developmental programming of other tissues. Overall, the placenta is responsive to environmental factors and uses a number of different strategies to adapt its phenotype to help support fetal growth during adverse intrauterine conditions. It is, therefore, not just a passive conduit for nutrient transfer to the fetus but alters its nutrient supply capacity dynamically to optimise fetal nutrient acquisition. Thus, the placental epigenome provides both a memory of environmental conditions experienced during development and an index of the future well being of the offspring.

特别是,出生时的宫内生长模式和大小决定了许多物种以后生命中组织的结构和功能,这对人类中成人发病的生殖疾病的发病率具有重要影响。在哺乳动物中,子宫内生长的主要决定因素是胎盘的营养供应,而营养供应又取决于胎盘的大小、形态、运输特性和内分泌功能。然而,与体细胞组织相比,我们对胎盘的发育程序知之甚少。本文综述了胎盘表型的表观遗传调控,特别强调了羊胎盘的营养转移能力和显示导致其他组织发育规划的环境因素。总的来说,胎盘对环境因素有反应,并使用许多不同的策略来适应其表型,以帮助在不利的宫内条件下支持胎儿生长。因此,它不仅是一个被动的管道,将营养物质转移到胎儿,但改变其营养供应能力动态优化胎儿的营养获取。因此,胎盘表观基因组提供了发育过程中所经历的环境条件的记忆和后代未来健康的指标。
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引用次数: 0
Introduction of the George Eric Lamming Memorial Lecture. 乔治·埃里克·兰明纪念讲座简介。
Keith Inskeep
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引用次数: 0
Mechanisms and pathobiology of ovulation. 排卵的机制和病理生物学。
Pub Date : 2010-01-01 DOI: 10.7313/upo9781907284991.017
W J Murdoch, C J Murphy, E A Van Kirk, Y Shen

The ovulatory process is extraordinary in that it constitutes a hormone-induced injury. Gonadotropin delivered via the follicular vascular wreath stimulates secretion of plasminogen activator by contiguous ovarian surface epithelial cells. A consequent elevation in interstitial plasmin activates collagenases and cleaves tumor necrosis factor alpha from its anchors on endothelium. Collagen fibril degradation and cellular death at the apex of the preovulatory follicle are hallmarks of impending ovulation. Follicular contractions rupture the weakened fabric at the apex, and the ovum, which has been disconnected from the underlying granulosa, is expelled; these components of the cascade are prostaglandin-mediated. Ovulation is required for fertility; unfortunately, it imparts a cancer risk to the ovarian surface epithelium. DNA-damaging reactive oxygen species are generated by inflammatory cells attracted into the vicinity of the ovulatory stigma. An ischemia-reperfusion flux coincident with ovulation and wound repair also contributes to genotoxicity. Potentially mutagenic lesions in DNA are normally reconciled by TP53 tumor suppressor-dependent cell-cycle arrest and base excision repair mechanisms; it is a unifocal escape that could be problematic. Epithelial ovarian cancer is a deadly insidious disease because it typically remains asymptomatic until it has metastasized to vital abdominal organs.

排卵过程是不寻常的,因为它构成了激素诱导的损伤。促性腺激素经卵泡血管环输送,刺激相邻卵巢表面上皮细胞分泌纤溶酶原激活剂。随之而来的间质纤溶酶升高激活胶原酶,并使肿瘤坏死因子α脱离其锚定在内皮上的位置。排卵前卵泡顶端的胶原纤维降解和细胞死亡是即将排卵的标志。卵泡收缩使顶端脆弱的组织破裂,与下层颗粒分离的卵子被排出;级联反应的这些组成部分是前列腺素介导的。排卵是生育所必需的;不幸的是,它会给卵巢表面上皮细胞带来癌症风险。损伤dna的活性氧是由炎性细胞吸引到排卵期柱头附近产生的。与排卵和伤口修复相一致的缺血再灌注通量也有助于遗传毒性。DNA中潜在的诱变损伤通常通过TP53肿瘤抑制因子依赖的细胞周期阻滞和碱基切除修复机制进行调解;这是一种可能有问题的单一逃逸。上皮性卵巢癌是一种致命的潜伏性疾病,因为它通常在转移到重要的腹部器官之前没有症状。
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引用次数: 16
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