Three regions of the ruminant oviduct play different roles in the progress of sperm: the uterotubal junction, isthmus, and ampulla. The uterotubal junction acts as a point of selection of sperm, requiring that sperm are progressively motile and express specific proteins in order to enter the oviduct. The isthmus stores sperm, preserving motility and viability until ovulation. Sperm are stored in the isthmus by binding to its mucosal epithelium. In bovine sperm, binding to the oviductal epithelium is promoted by proteins that are secreted by the seminal vesicles and coat the heads of sperm by associating with plasma membrane phospholipids. Putative oviductal receptors for the seminal vesicle proteins are members of the annexin protein family. Release of sperm from the storage site in the isthmus is gradual, which serves to ensure that sperm in the proper physiological state reach the oocytes at the appropriate moment and also to reduce incidence of polyspermic fertilization. The ampulla supports fertilization and may participate in guiding sperm toward the eggs. Further studies are needed to improve our understanding of the interactions between sperm and the female reproductive tract, in order to develop means to improve fertility in ruminants.
Three regions of the ruminant oviduct play different roles in the progress of sperm: the uterotubal junction, isthmus, and ampulla. The uterotubal junction acts as a point of selection of sperm, requiring that sperm are progressively motile and express specific proteins in order to enter the oviduct. The isthmus stores sperm, preserving motility and viability until ovulation. Sperm are stored in the isthmus by binding to its mucosal epithelium. In bovine sperm, binding to the oviductal epithelium is promoted by proteins that are secreted by the seminal vesicles and coat the heads of sperm by associating with plasma membrane phospholipids. Putative oviductal receptors for the seminal vesicle proteins are members of the annexin protein family. Release of sperm from the storage site in the isthmus is gradual, which serves to ensure that sperm in the proper physiological state reach the oocytes at the appropriate moment and also to reduce incidence of polyspermic fertilization. The ampulla supports fertilization and may participate in guiding sperm toward the eggs. Further studies are needed to improve our understanding of the interactions between sperm and the female reproductive tract, in order to develop means to improve fertility in ruminants.
Endogenous retroviruses (ERVs) are present in the genome of all vertebrates and are remnants of ancient exogenous retroviral infections of the host germline transmitted vertically from generation to generation. Sheep betaretroviruses offer a unique model system to study the complex interaction between retroviruses and their host. The sheep genome contains 27 endogenous betaretroviruses (enJSRVs) related to the exogenous and pathogenic Jaagsiekte sheep retrovirus (JSRV), the causative agent of a transmissible lung cancer in sheep. The enJSRVs can protect their host against JSRV infection by blocking early and late steps of the JSRV replication cycle. In the female reproductive tract, enJSRVs are specifically expressed in the uterine luminal and glandular epithelia as well as in the conceptus (embryo and associated extraembryonic membranes) trophectoderm and in utero loss-of-function experiments found the enJSRVs envelope (env) to be essential for conceptus elongation and trophectoderm growth and development. Collectively, available evidence in sheep and other mammals indicate that ERVs coevolved with their hosts for millions of years and were positively selected for biological roles in genome plasticity and evolution, protection of the host against infection of related pathogenic and exogenous retroviruses, and placental development.
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.
The purpose of the review is to describe the factors that affect fertility in domestic water buffalo (Bubalus bubalis) and the techniques that enable an improvement in reproductive performance. On Italian and Latin American farms where natural mating is practiced and bulls are always present in the herd, the inter-calving interval is approximately 400 days and the culling rate is lower than 15%. The buffalo has a tendency for seasonal reproductive activity. Reproduction is favoured when there is a decrease in day length. Ovarian activity stops if conception does not occur within 3 to 5 ovarian cycles. It is important, therefore, that appropriate management of the transition period is practiced, particularly with respect to the hygienic conditions of the uterus. In tropical countries located north of the equator, feed deficiencies and heat stress are considered the main factors that lead to poor fertility in the summer. In Pakistan, for example, the increase in body condition score during the autumn was associated with the commencement of the breeding season in buffaloes. Anoestrus is observed also in Italy, however, where the average daily temperature during the same period is 13.5 to 23.5 degrees C and feeding is constant throughout the year. The only common element between the two areas is the progressive increase in daylight hours between April and June and the day length greater than 12 hours up to September. In Italian herds that apply an out-of-season breeding strategy, an improvement in fertility (measured as the percentage of corpora lutea corresponding to subsequent pregnancy) is observed when water pools are present on the farm. This demonstrates that an improvement in environmental conditions reduces the incidence of embryonic mortality and/or abnormal cycles. To summarize, in the absence of serious nutritional problems, an improvement in environmental conditions increases fertility in buffalo.
Estrous synchronization and timed ovulation programs that permit AI at a predetermined time (timed AI) rather than as determined by detection of spontaneous estrus are requisite for increased adoption of AI in the beef cattle industry. In the past two decades, significant progress has been achieved in developing programs that synchronize ovulation to address this need. While this progress has been driven by a multitude of fundamental discoveries in reproductive biology, the greatest impact in the past two decades has been the result of enhanced understanding of the pattern of ovarian follicle growth in cattle and development of technologies to coordinate growth and ovulation of the dominant follicle. At present, estrous synchronization programs that result in acceptable timed AI pregnancy rates are available for beef cattle. The capacity to control growth of the dominant follicle and evaluate the impact of various approaches on fertility has resulted in greater understanding of the factors that influence maturity of ovulatory follicles. Modifications to the standard industry breeding programs, with the aim of lengthening and/or increasing the gonadotropic stimulus and estradiol production by preovulatory follicles, have been shown to substantially increase timed AI pregnancy rate in beef cattle. Associations between characteristics of follicular development and fertility have surfaced from application of estrous and ovulation synchronization technologies and led to investigation of the fundamental mechanisms that underlie these relationships.
Circannual clocks drive rhythms in reproduction and many other seasonal characteristics but the underlying control of these long-term oscillators remains a mystery. Now, we propose that circannual timing involves mechanisms that are integral to the ontogenetic life-history programme where annual transitions are generated by cell birth, death and tissue regeneration throughout the life cycle--the histogenesis hypothesis. The intrinsic cycle is then timed by cues from the environment. The concept is that in specific sites in the brain, pituitary and peripheral tissues, residual populations of progenitor cells (adult stem cells) synchronously initiate a phase of cell division to begin a cycle. The progeny cells then proliferate, migrate and differentiate, providing the substrate that drives physiological change over long time-spans (e.g. summer/winter); cell death may be required to trigger the next cycle. We have begun to characterise such a tissue-based timer in our Soay sheep model focusing on the pars tuberalis (PT) of the pituitary gland and the sub-ventricular zone of the mediobasal hypothalamus (MBH) as potential circannual pacemakers. The PT is of special interest because it is a melatonin-responsive tissue containing undifferentiated cells, strategically located at the gateway between the brain and pituitary gland. The PT also governs long-photoperiod activation of thyroid hormone dependant processes in the MBH required for neurogenesis. In sheep, exposure to long photoperiod markedly activates BrDU-labelled cell proliferation in the PT and MBH, and acts to entrain the circannual reproductive cycle. Variation in expression and co-ordination of multiple tissue timers may explain species differences in circannual rhythmicity. This paper is dedicated to the memory of Ebo Gwinner.
The New Zealand (NZ) economy and its dairy industry are sensitive to global consumer perception of farming practices used to generate milk products because milk exports account for > 25% of national export earnings and > 90% of milk produced is exported as products. Astute management of product image and market risk is, therefore, important for the viability of the industry and country. More than 95% of milk produced in NZ comes from strictly seasonal, pasture-based systems, with associated constraints on reproductive performance. Increasing herd sizes, operational changes and genetic selection priorities have further challenged dairy farmers to achieve optimal levels of herd fertility. Reproductive management practices have developed to address the need to maintain a 365-day inter-calving interval, essentially through maximizing the number of cyclic cows during the breeding period and minimizing the duration of the seasonal calving period. Aspects of the hormonal interventions developed and routinely used to achieve these objectives have been the subject of product quality and market risk concerns forcing the industry to explore alternative ways of achieving reproductive performance goals. One approach has been to exploit the inherently high level of fertility in NZ dairy herds. This approach has seen the inclusion of fertility-related traits in the national genetic evaluation system to prevent further decline in genetic fertility. More recently, a nationally coordinated extension program has been adopted to support farmers and their advisors to identify, prioritize and improve on key management areas for incremental gains in herd reproductive performance. Advances in automation and bio-sensing are yet to make a significant impact, but remain potentially valuable additions in supporting the dairy farmer to manage the areas having the largest effects on reproductive performance.
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.
The formation of primordial follicles to establish a reservoir of resting follicles and the gradual depletion of that reservoir to provide a succession of growing follicles are key to female fertility, but little is known about the regulation of these early stages of follicular development. This review summarizes the efforts of our laboratory to elucidate these critical processes in cattle. Primordial follicles first appear in fetal ovaries around the end of the first trimester of pregnancy (Day 90), during a decline in fetal ovarian production of estradiol and progesterone. In ovarian cortical pieces from 90 to 120-day-old fetuses, follicles form in vitro and estradiol or progesterone inhibits follicle formation, whereas the non-aromatizable androgen 5alpha-dihydrotestosterone (DHT) does not. Newly formed bovine follicles are not capable of activating within 2 days in vitro, but they can acquire the capacity to activate during a longer culture; estradiol and progesterone inhibit the acquisition of their capacity to activate. When primordial follicles first form in cattle, their oocytes are not yet in meiotic arrest and acquisition of competence to activate is correlated with their progression to meiotic arrest at the diplotene stage of first prophase. After they acquire the competence to activate, bovine primordial follicles can be stimulated to activate in vitro by insulin or kit ligand, whereas anti-Mullerian hormone (AMH) is inhibitory. Although few follicles progress to the secondary stage in vitro, addition of testosterone or vascular endothelial growth factor (VEGF) dramatically increased the incidence of that transition. Regulation of the earliest stages of follicular development is complex and far from understood; better understanding could lead to new interventions to enhance fertility.
Development of the corpus luteum (CL) in ruminants occurs in a rapid and time-dependent manner within 1 week after ovulation, with morphologic and biochemical changes in the cells of the theca interna and granulosa cells of the preovulatory follicle. These changes involve luteinisation of steroidogenic cells and angiogenesis to establish normal luteal function (progesterone secretion). The CL is composed of a large number of vascular endothelial cells, large and small steroidogenic luteal cells, smooth muscle cells, pericytes, fibrocytes and immune cells, indicating that the CL is a heterogeneous tissue. Moreover, the CL produces and secretes growth factors (fibroblast growth factor, vascular endothelial growth factor and insulin-like growth factor), vasoactive factors (nitric oxide, angiotensin II and endothelin-1), steroids (progesterone is important for its own production), oxytocin and prostaglandins (PGF2alpha and PGE2) to regulate luteal formation and development. Clearly, the main function of the CL is to produce progesterone, which is a prerequisite for survival of the embryo, implantation and maintenance of pregnancy. Inadequate luteinisation and angiogenesis during the early luteal phase results in poor progesterone secretion and causes compromised embryo development and reduced fertility. Secretion of adequate amounts of progesterone during luteal development requires "precise luteinisation" of theca and granulosa cells to form luteal cells, neovascularization, and the establishment of a blood supply (angiogenesis). PGF2alpha in the developing CL acts as a local regulator to enhance progesterone secretion directly and indirectly by stimulating angiogenic factors, VEGF and FGF2. The preceding role of PGF2alpha may explain why the developing CL does not acquire luteolytic capacity until several days following ovulation. The balance between luteotrophic and luteolytic factors as well as stimulation and inhibition of angiogenic factors during luteal formation, development and maintenance can have a profound effect on the fate of the CL.
In cattle we have noted that the antral follicle count (AFC, follicles > or = 3 mm in diameter) varies greatly among animals (from 5 to 50), is repeatable within animals, and is highly correlated with the total number of healthy follicles in ovaries. Also, animals with low AFC have higher serum concentrations of FSH and LH, but lower concentrations of Anti-Mullerian Hormone, progesterone and androgens than animals with high AFC. We have investigated the effect of maternal environment during gestation on their offspring AFC by restricting maternal nutrition to 60% of maintenance requirements (compared with 100% in controls) during the first third of gestation. Calves born to nutritionally restricted mothers had 60% lower AFC compared with calves born to mothers fed control diets. In other studies we have evidence to indicate that fertility may be compromised in animals with low AFC due to effects on oocytes, progesterone and the endometrium compared with animals with high AFC. To examine this directly we assessed AFC in post-partum dairy cows and found that cows with a high AFC had higher pregnancy rates, shorter calving to conception intervals and received fewer services during the breeding season compared with cows with a low AFC. In addition, the high variation in follicle numbers in adults may not only be reflective of reproductive disorders and suboptimal fertility, but there is evidence to indicate that it may be associated with alterations in the function of other non-reproductive systems (e.g. cardiovascular) that may have profound effects on the animal's health and welfare.
In the current review the main fundamental biological differences in reproductive function between Bos taurus and Bos indicus cattle are discussed. Breed differences regarding puberty, estrous cycle patterns, estrous behavior, acquisition of ovulatory capacity, ovarian structures and reproductive hormones are presented. The main physiological differences that Bos indicus cattle present relative to Bos taurus cattle include: delayed age at puberty; higher circulating concentrations of hormones such as estradiol, progesterone, insulin and IGF-I, despite having smaller ovulatory follicle size and corpora lutea; greater population of small follicles and smaller size of the dominant follicle at deviation; and greater sensitivity of follicles to gonadotropins. Knowledge of the differences between Bos indicus and Bos taurus breeds help explain different management procedures and responses to hormonal treatments associated with artificial insemination, ovarian superstimulation, and in vivo and in vitro embryo production.
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.
Reindeer (Rangifer tarandus tarundus) are the only cervids indigenous to the arctic environment. In Alaska, reindeer are a recognized agricultural species and an economic mainstay for many native populations. Traditionally raised in extensive free-ranging systems, a recent trend toward intensive farming requires a more in-depth knowledge of reproductive management. Reported gestation length in reindeer varies, ranging from 198 to 229 d in studies performed at the University of Alaska Fairbanks. A switchback study that manipulated only breeding date demonstrated a mean increase in gestation length of 8.5 d among females bred early in the season. The negative correlation between conception date and gestation length is consistent with reindeer research at other locations and reports of variable gestation length in a growing number of domestic and non-domestic species. This paper reviews the phenomenon in reindeer and discusses some of the factors known to affect gestation length as well as possible areas for future research.
The placenta provides the means for nutrient transfer from the mother to the fetus, waste transfer from the fetus to the mother, protection of the fetus from the maternal immune system, and is an active endocrine organ. While many placental functions have been defined and investigated, assessing the function of specific genes expressed by the placenta has been problematic, since classical ablation-replacement methods are not feasible with the placenta. The pregnant sheep has been a long-standing animal model for assessing in vivo physiology during pregnancy, since surgical placement of indwelling catheters into both maternal and fetal vasculature has allowed the assessment of placental nutrient transfer and utilization, as well as placental hormone secretion, under unanesthetized-unstressed steady state sampling conditions. However, in ruminants the lack of well-characterized trophoblast cell lines and the inefficiency of creating transgenic pregnancies in ruminants have inhibited our ability to assess specific gene function. Recently, sheep and cattle primary trophoblast cell lines have been reported, and may further our ability to investigate trophoblast function and transcriptional regulation of genes expressed by the placenta. Furthermore, viral infection of the trophoectoderm layer of hatched blastocysts, as a means for placenta-specific transgenesis, holds considerable potential to assess gene function in the ruminant placenta. This approach has been used successfully to "knockdown" gene expression in the developing sheep conceptus, and has the potential for gain-of-function experiments as well. While this technology is still being developed, it may provide an efficient approach to assess specific gene function in the ruminant placenta.
Sheep are seasonal breeders and reproductive status is controlled by photoperiod. Recent recognition of the significant role for kisspeptin and gonadotropin inhibitory hormone (GnIH) in the regulation of gonadotropin releasing hormone (GnRH) cells has provided a new perspective in the seasonal regulation of reproductive activity. Virtually all kisspeptin cells express estrogen receptors and kisspeptin is a potent stimulator of GnRH secretion. Thus, kisspeptin cells provide a conduit by which changes in estrogen feedback effects may be exerted upon GnRH cells. Changes in the activity of kisspeptin cells with season indicate a major role in the seasonal changes in reproductive activity in the ewe. GnIH is an inhibitor of reproductive function and there is mounting evidence that changing activity of this system is also an important determinant of reproductive status. Reciprocal changes in kisspeptin and GnIH activity explain seasonal changes in the function of GnRH cells.
In order to achieve fertilization sperm cells, first need to successfully interact with the zona pellucida. To this end, the sperm surface is extensively remodeled during capacitation and the resulting sperm cells also possess hyperactivated motility. Together, this serves to mediate optimal recognition of the zona pellucida in the oviduct or after in vitro fertilization incubations (primary zona pellucida binding). When the sperm cell attaches to the zona pellucida, it will be triggered to undergo the acrosome reaction which allows the hyperactivated motile sperm cell to drill through the zona pellucida (secondary zona pellucida binding coinciding with sequential local zona pellucida digestion and rebinding). After successful zona penetration, some sperm cells may enter the perivitelline space. This delaying strategy of the oocyte allows only one sperm cell at a given time to bind and fuse with the oocyte (fertilization) and thus minimizes the risk of polyspermy. The fertilization fusion between the oocyte and the first sperm cell is immediately followed by a polyspermic fertilization block, in which the content of the oocyte's cortical granules is released into the perivitelline space. The cortical reaction blocks further sperm-oocyte fusion either by sticking at the oolemma or by the induction of a biochemical reaction of the zona pellucida (zona pellucida hardening). The cortical reaction thus blocks sperm-zona pellucida binding and/or sperm-zona pellucida penetration. This review summarizes the current understanding of sperm-zona pellucida interactions in relation to mammalian fertilization. The lack of knowledge about sperm-zona pellucida binding in ruminants will be critically discussed.
The establishment and maintenance of pregnancy in lactating dairy cows is a complex biological event that is influenced by a multitude of factors, from the reproductive biology of the cow to managerial aspects of the dairy farm. It is often mentioned in the scientific literature that fertility in dairy cows has declined concurrent with major advances in milk production. Some of this decline is attributed to the negative genetic correlation between milk production and reproduction. In the United States, yearly production per cow has increased steadily at a rate of 1.3% in the last decade and it is likely that this trend will continue in the years to come. At this rate, the average cow in the United States will be producing over 14 tons of milk per year in 2050 and technologies will have to be developed to allow these cows to reproduce to maintain the sustainability of dairy production. Despite high production, it is not uncommon for dairy herds with rolling herd averages for milk yield above 11,000 kg to overcome the challenges of reproduction and obtain satisfactory reproductive performance. Among other things, those herds have been able to mitigate some of the mechanisms that suppress reproduction in dairy cows such as extended postpartum anovulatory period, poor estrous detection, low pregnancy per insemination and, to a lesser extent, the high pregnancy loss. The success of those farms comes from an integrated approach to fertility that includes adequate cow comfort, elaborated transition cow management and nutrition, aggressive postpartum health monitoring program with preventative and curative measures to mitigate the negative effects of diseases on reproduction, and a sound reproductive program that includes manipulation of the ovarian cycle to allow for increased insemination rate. More recently, introduction of fertility traits in selection programs have created new opportunities for improved reproduction without neglecting economically important production traits.
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).
