Embryonic morphology observation and HOXC8 gene expression in crest cushions of Chinese Crested duck.

The Chinese Crested duck (Fengtou duck) reappeared in China recently. Along with white feathers and a black bill and feet, the Fengtou duck has a high feather crest. This breed can be used for ornamental purposes or as a model organism; however, little is known about the genetic basis and development of its distinct morphological features. In this study, we observed the skull and feather crest of Fengtou duck in the embryonic stages. As a result, the protuberances of the head integument could be clearly observed at embryonic stage E9, and small perforations in the skull were first visible at E13 and were clearer at E15. Besides, intracranial fat in a small number of individuals was found starting at E15, and a small number of osteophytes was found at E18. In addition, hematoxylin-eosin (HE) and Oil Red-O staining of the crest cushion and intracranial tissue revealed fat tissue accumulation. Previous studies demonstrated that homeobox c8 (HOXC8) played a critical role in chicken crest formation. Here, we cloned the HOXC8 from Fengtou duck and determined that its transcript was highly expressed in the crest cushion; moreover, HOXC8 was detected in this tissue with a molecular weight of 38 kDa in the Fengtou duck. In conclusion, embryos of Fengtou duck have different small protuberances and perforations in the skull, including accumulation of intracranial tissue and osteophytes in some cases. Furthermore, HOXC8 may regulate the formation of the crest. These findings provide novel insight into the ontogenesis of the crest cushion in crested ducks and a basis for future studies on their evolutionary origins.

Embryonic effects of an environmentally relevant PCB mixture in the domestic chicken.

Studies were conducted to develop methods to assess the effects of a complex mixture of polychlorinated biphenyls (PCBs) in the domestic chicken (Gallus domesticus). Treatments were administered by egg injection to compare embryonic effects of an environmentally relevant PCB congener mixture in the domestic chicken over a range of doses. Chicken eggs were injected with the PCB mixture with a profile similar to that found in avian eggs collected on the upper Hudson River, New York, USA, at doses that spanned 0 to 98 µg/g egg. Eggs were hatched in the laboratory to ascertain hatching success. In the domestic chicken, the median lethal dose was 0.3 µg/g. These data demonstrate adverse effects of an environmentally relevant PCB mixture and provide the basis for further work using in vitro and other models to characterize the potential risk to avian populations. Environ Toxicol Chem 2018;37:2513-2522. © 2018 SETAC.

Pre-implantation Development of Domestic Animals.

During the first days following fertilization, cells of mammalian embryo gradually lose totipotency, acquiring distinct identity. The first three lineages specified in the mammalian embryo are pluripotent epiblast, which later gives rise to the embryo proper, and two extraembryonic lineages, hypoblast (also known as primitive endoderm) and trophectoderm, which form tissues supporting development of the fetus in utero. Most of our knowledge regarding the mechanisms of early lineage specification in mammals comes from studies in the mouse. However, the growing body of evidence points to both similarities and species-specific differences. Understanding molecular and cellular mechanisms of early embryonic development in nonrodent mammals expands our understanding of basic mechanisms of differentiation and is essential for the development of effective protocols for assisted reproduction in agriculture, veterinary medicine, and for biomedical research. This review summarizes the current state of knowledge on key events in epiblast, hypoblast, and trophoblast differentiation in domestic mammals.

Genomic determinants of epidermal appendage patterning and structure in domestic birds.

Variation in regional identity, patterning, and structure of epidermal appendages contributes to skin diversity among many vertebrate groups, and is perhaps most striking in birds. In pioneering work on epidermal appendage patterning, John Saunders and his contemporaries took advantage of epidermal appendage diversity within and among domestic chicken breeds to establish the importance of mesoderm-ectoderm signaling in determining skin patterning. Diversity in chickens and other domestic birds, including pigeons, is driving a new wave of research to dissect the molecular genetic basis of epidermal appendage patterning. Domestic birds are not only outstanding models for embryonic manipulations, as Saunders recognized, but they are also ideal genetic models for discovering the specific genes that control normal development and the mutations that contribute to skin diversity. Here, we review recent genetic and genomic approaches to uncover the basis of epidermal macropatterning, micropatterning, and structural variation. We also present new results that confirm expression changes in two limb identity genes in feather-footed pigeons, a case of variation in appendage structure and identity.

Invited review: Pre- and postnatal adipose tissue development in farm animals: from stem cells to adipocyte physiology.

Both white and brown adipose tissues are recognized to be differently involved in energy metabolism and are also able to secrete a variety of factors called adipokines that are involved in a wide range of physiological and metabolic functions. Brown adipose tissue is predominant around birth, except in pigs. Irrespective of species, white adipose tissue has a large capacity to expand postnatally and is able to adapt to a variety of factors. The aim of this review is to update the cellular and molecular mechanisms associated with pre- and postnatal adipose tissue development with a special focus on pigs and ruminants. In contrast to other tissues, the embryonic origin of adipose cells remains the subject of debate. Adipose cells arise from the recruitment of specific multipotent stem cells/progenitors named adipose tissue-derived stromal cells. Recent studies have highlighted the existence of a variety of those cells being able to differentiate into white, brown or brown-like/beige adipocytes. After commitment to the adipocyte lineage, progenitors undergo large changes in the expression of many genes involved in cell cycle arrest, lipid accumulation and secretory functions. Early nutrition can affect these processes during fetal and perinatal periods and can also influence or pre-determinate later growth of adipose tissue. How these changes may be related to adipose tissue functional maturity around birth and can influence newborn survival is discussed. Altogether, a better knowledge of fetal and postnatal adipose tissue development is important for various aspects of animal production, including neonatal survival, postnatal growth efficiency and health.

Stem cells and regenerative medicine in domestic and companion animals: a multispecies perspective.

Since their original isolation, the majority of the work on embryonic stem cells (ESC) has been carried out in mice. While the mouse is an outstanding model for basic research, it also has considerable limitations for translational work, especially in the area of regenerative medicine. This is due to a combination of factors that include physiological and size differences when compared to humans. In contrast, domestic animal species, such as swine, and companion animal species, such as dogs, provide unique opportunities to develop regenerative medicine protocols that can then be utilized in humans. Unfortunately, at present, the state of knowledge related to, and availability of, ESC from domestic animals vary among species such as pig, horse, dog and cat, and without exception lags significantly behind the mouse and human. It is clear that much still needs to be discovered. The 'stem cell-like' cell lines being reported are still not satisfactorily used in regenerative medicine, due to reasons such as heterogeneity and chromosomal instability. As a result, investigators have searched for alternate source of cells that can be used for regenerative medicine. This approach has uncovered a range of adult stem cells and adult progenitor cells that have utility in both human and veterinary medicine. Here, we review a range of stem cells, from ESC to induced pluripotent stem cells, and discuss their potential application in the field of regenerative medicine.

Preimplantation embryo metabolism and culture systems: experience from domestic animals and clinical implications.

Despite advantages of in vitro embryo production in many species, widespread use of this technology is limited by generally lower developmental competence of in vitro derived embryos compared to in vivo counterparts. Regardless, in vivo or in vitro gametes and embryos face and must adjust to multiple microenvironments especially at preimplantation stages. Moreover, the embryo has to be able to further adapt to environmental cues in utero to result in the birth of live and healthy offspring. Enormous strides have been made in understanding and meeting stage-specific requirements of preimplantation embryos, but interpretation of the data is made difficult due to the complexity of the wide array of culture systems and the remarkable plasticity of developing embryos that seem able to develop under a variety of conditions. Nevertheless, a primary objective remains meeting, as closely as possible, the preimplantation embryo requirements as provided in vivo. In general, oocytes and embryos develop more satisfactorily when cultured in groups. However, optimization of individual culture of oocytes and embryos is an important goal and area of intensive current research for both animal and human clinical application. Successful culture of individual embryos is of primary importance in order to avoid ovarian superstimulation and the associated physiological and psychological disadvantages for patients. This review emphasizes stage specific shifts in embryo metabolism and requirements and research to optimize in vitro embryo culture conditions and supplementation, with a view to optimizing embryo culture in general, and culture of single embryos in particular.

Therio-ontology: a personal view of 40 years of farm animal embryo form and function.

The main purpose of this autobiographical reminiscence of 40 years of embryo research is to provide young theriogenologists with a firsthand account of how career development can depend strongly on early influences that become modified by changing circumstances. With no intention of being didactic, I hope that my experience of coping with enormous changes in techniques and attitudes may be of use to some of those embarking on a further 40 years of change of at least equal enormity.

From "ES-like" cells to induced pluripotent stem cells: a historical perspective in domestic animals.

Pluripotent stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) provide great potential as cell sources for gene editing to generate genetically modified animals, as well as in the field of regenerative medicine. Stable, long-term ESCs have been established in laboratory mouse and rat; however, isolation of true pluripotent ESCs in domesticated animals such as pigs and dogs have been less successful. Initially, domesticated animal pluripotent cell lines were referred to as "embryonic stem-like" cells owing to their similar morphologic characteristics to mouse ESCs, but accompanied by a limited ability to proliferate in vitro in an undifferentiated state. That is, they shared some but not all the characteristics of true ESCs. More recently, advances in reprogramming using exogenous transcription factors, combined with the utilization of small chemical inhibitors of key biochemical pathways, have led to the isolation of iPSCs. In this review, we provide a historical perspective of the isolation of various types of pluripotent stem cells in domesticated animals. In addition, we summarize the latest progress and limitations in the derivation and application of iPSCs.

Vitrification in human and domestic animal embryology: work in progress.

According to the analysis of papers published in major international journals, rapidly increasing application of vitrification is one of the greatest achievements in domestic animal and especially human embryology during the first decade of our century. This review highlights factors supporting or hampering this progress, summarises results achieved with vitrification and outlines future tasks to fully exploit the benefits of this amazing approach that has changed or will change many aspects of laboratory (and also clinical) embryology. Supporting factors include the simplicity, cost efficiency and convincing success of vitrification compared with other approaches in all species and developmental stages in mammalian embryology, while causes that slow down the progress are mostly of human origin: inadequate tools and solutions, superficial teaching, improper application and unjustified concerns resulting in legal restrictions. Elimination of these hindrances seems to be a slower process and more demanding task than meeting the biological challenge. A key element of future progress will be to pass the pioneer age, establish a consensus regarding biosafety requirements, outline the indispensable features of a standard approach and design fully-automated vitrification machines executing all phases of the procedure, including equilibration, cooling, warming and dilution steps.

Pluripotent cells in farm animals: state of the art and future perspectives.

Pluripotent cells, such as embryonic stem (ES) cells, embryonic germ cells and embryonic carcinoma cells are a unique type of cell because they remain undifferentiated indefinitely in in vitro culture, show self-renewal and possess the ability to differentiate into derivatives of the three germ layers. These capabilities make them a unique in vitro model for studying development, differentiation and for targeted modification of the genome. True pluripotent ESCs have only been described in the laboratory mouse and rat. However, rodent physiology and anatomy differ substantially from that of humans, detracting from the value of the rodent model for studies of human diseases and the development of cellular therapies in regenerative medicine. Recently, progress in the isolation of pluripotent cells in farm animals has been made and new technologies for reprogramming of somatic cells into a pluripotent state have been developed. Prior to clinical application of therapeutic cells differentiated from pluripotent stem cells in human patients, their survival and the absence of tumourigenic potential must be assessed in suitable preclinical large animal models. The establishment of pluripotent cell lines in farm animals may provide new opportunities for the production of transgenic animals, would facilitate development and validation of large animal models for evaluating ESC-based therapies and would thus contribute to the improvement of human and animal health. This review summarises the recent progress in the derivation of pluripotent and reprogrammed cells from farm animals. We refer to our recent review on this area, to which this article is complementary.

Utility of animal models for human embryo culture development: domestic species.

Although the mouse is widely used as an animal model for the in vitro culture of human preimplantation embryos, arguments can be made for the validity of many domestic species as more relevant, applicable models of early human development. Here, we describe the advantages and disadvantages of domestic species as animal models to develop in vitro culture methods and conditions that successfully support human embryo development. In vitro embryo culture procedures are described and compared between species for the bovine, ovine, caprine, and porcine species.

Skeletal muscle proteomics in livestock production.

Proteomics allows studying large numbers of proteins, including their post-translational modifications. Proteomics has been, and still are, used in numerous studies on skeletal muscle. In this article, we focus on its use in the study of livestock muscle development and meat quality. Changes in protein profiles during myogenesis are described in cattle, pigs and fowl using comparative analyses across different ontogenetic stages. This approach allows a better understanding of the key stages of myogenesis and helps identifying processes that are similar or divergent between species. Genetic variability of muscle properties analysed by the study of hypertrophied cattle and sheep are discussed. Biological markers of meat quality, particularly tenderness in cattle, pigs and fowl are presented, including protein modifications during meat ageing in cattle, protein markers of PSE meat in turkeys and of post-mortem muscle metabolism in pigs. Finally, we discuss the interest of proteomics as a tool to understand better biochemical mechanisms underlying the effects of stress during the pre-slaughter period on meat quality traits. In conclusion, the study of proteomics in skeletal muscles allows generating large amounts of scientific knowledge that helps to improve our understanding of myogenesis and muscle growth and to control better meat quality.

Developmental programming: the concept, large animal models, and the key role of uteroplacental vascular development.

Developmental programming refers to the programming of various bodily systems and processes by a stressor of the maternal system during pregnancy or during the neonatal period. Such stressors include nutritional stress, multiple pregnancy (i.e., increased numbers of fetuses in the gravid uterus), environmental stress (e.g., high environmental temperature, high altitude, prenatal steroid exposure), gynecological immaturity, and maternal or fetal genotype. Programming refers to impaired function of numerous bodily systems or processes, leading to poor growth, altered body composition, metabolic dysfunction, and poor productivity (e.g., poor growth, reproductive dysfunction) of the offspring throughout their lifespan and even across generations. A key component of developmental programming seems to be placental dysfunction, leading to altered fetal growth and development. We discuss various large animal models of developmental programming and how they have and will continue to contribute to our understanding of the mechanisms underlying altered placental function and developmental programming, and, further, how large animal models also will be critical to the identification and application of therapeutic strategies that will alleviate the negative consequences of developmental programming to improve offspring performance in livestock production and human medicine.

Intracytoplasmic sperm injection in livestock species: an update.

Intracytoplasmic sperm injection (ICSI) is a powerful technique in the field of assisted reproduction (ART) and provides exciting opportunities for studying the basic mechanisms of fertilization and early embryo development. Nevertheless, its application in agriculture and conservation biology has been greatly hampered by the low success rate reported for this method in respect of economically important species. Specifically, the rates of blastocyst formation and live newborn are greatly reduced when zygotes are generated by ICSI. Except for humans, ICSI remains a low efficiency technology in comparison with alternatives such as in vitro fertilization (IVF) and its application is less widespread. In this paper, we discuss the present status, applications and factors affecting ICSI in pigs and other species.

Recent progress in embryonic stem cell research and its application in domestic species.

Many reports described cell lines derived in domestic species, which presented several important features typical of embryonic stem cells (ESCs). Such features unfortunately did not include the capacity to generate germ-line chimeras, therefore limiting the possibility to use these cells as tools for the genetic manipulation. However, farm animal ESCs may still be useful for the generation of transgenic animals as usually have a self-renewal capacity more prolonged than normal primary cultures thus increasing the possibility to transform and select cells to be used as nucleus donors in cloning procedures. Farm animal ESCs may also be an excellent experimental model in pre-clinical trials, assessing the feasibility of cell therapy because of the close morphological and physiological resemblance to humans of species like the pig. However, the persistent lack of standard methods for the derivation, maintenance and characterization of ESCs in domestic species stimulated the search for alternatives. Embryonic germ cells may represent such an alternative. Indeed, these cells showed a higher plasticity than ESCs as contributed to embryonic development forming chimeric newborns but, as for ESCs, standardization is still far away and efficiency is very low. Recent results indicated spermatogonial stem cells as possible tools for germ-line genetic modifications with some proof of principle results already achieved. But, a real break through could arrive from the multipotent germ-line stem cells, virtually equivalent to ESC, derived from newborn and adult mouse testis.

Chromosome variation in the embryos of domestic animals.

Chromosome abnormalities in the embryos of domestic animals are mostly eliminated during development. De novo chromosome abnormalities in the embryos of domestic animals have been detected in a larger proportion of embryos produced by in vitro fertilization and somatic cell nuclear transfer than in those produced by natural mating or artificial insemination. The increased incidence of abnormalities in embryos produced in vitro provides evidence for an influence of the embryo production procedures on chromosome stability. Research strategies involving cytogenetics, molecular biology and reproductive biotechnologies hold the promise of yielding insight into the mechanisms underlying chromosome instability in embryos and the impact of the in vitro environment on the chromosome make-up of embryos.

Opportunities and challenges in applying genomics to the study of oogenesis and folliculogenesis in farm animals.

Ovarian oogenesis and folliculogenesis are complex and coordinated biological processes which require a series of events that induce morphological and functional changes within the follicle, leading to cell differentiation and oocyte development. In this context, the challenge of the researchers is to describe the dynamics of gene expression in the different compartments and their interactions during the follicular programme. In recent years, high-throughput arrays have become a powerful tool with which to compare the whole population of transcripts in a single experiment. Here, we review the challenges of applying genomics to this model in farm animal species. The first limitation lies in limited the availability of biological material, which makes the study of the follicle compartments (oocyte, granulosa cells and thecal cells) or early embryo much more difficult. The concept of observing all transcripts at once is very attractive but despite progress in sequencing, the genome annotation remains very incomplete in non-model species. Particularly, oogenesis and early embryo development relate to the high proportion of unknown expressed sequence tags. Then, it is important to consider post-transcriptional and translational regulation to understand the role of these genes. Ultimately, these new inferred insights will still have to be validated by functional approaches. In addition to in vitro or ex vivo functional approaches, both 'natural mutant' ewe models and RNA interference represent, at the moment, the best hope for functional genomics. Advances in our understanding of reproductive physiology should be facilitated by gene expression data exchange and translation into a better understanding of the underlying biological phenomena.