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Advances in biotechnology: new tools in future pig production for agriculture and biomedicine.

Biotechnology in livestock comprises an arsenal of reproductive biotechniques and molecular genetics. While molecular genetics are poorly developed in swine, reproductive techniques are more advanced and applied under field conditions. This review describes three selected examples of our own research to illustrate the implication of biotechnology in future pig reproduction. Sperm sexing technology is now available and can be used to generate piglets of the desired sex by IVF and ICSI. First studies also indicate satisfactory success rates following intrauterine insemination with sexed spermatozoa. Cloning technique and production of transgenic pigs require information about the regulation and time course of gene expression during in vitro production and pre-implantation development. Information on gene expression is scare in porcine embryos. With the exception of transcripts for the oestrogen receptor gene, no mRNA's from the activated porcine genome have been identified. Recent development of cDNA arrays might help to identify a larger amount of genes in single embryos. Remarkable progress has been made in organ transplantation technology. As the demand for human organs is increasing rapidly, the pig might serve as donor of xenotranplants, provided the transmission of zoonoses from the donor animal to the human recipient is prevented, donor organ anatomy and function are compatible, and immunological rejections (HAR, VAR MAC) can be overcome. The most promising strategy is the synthesis of human complement regulatory proteins in the pig. Transgenic pigs have been generated for hDAF or hCD 46 and their hearts have been transplanted into non human primates for up to 90 days. HCD 59 driven by CMV promotor provides significant protection against HAR at least under in vitro and in vivo conditions. Current studies indicate a temporary use in patients within the next 3-5 years. As the microinjection technology to produce transgenic offspring is time consuming and very expensive, nuclear transfer technology provides a possibility for multiplication without going through the germ line with recombination effects. A functional nuclear transfer system will be crucial for xenotransplantation as it is anticipated that the expression of several transgenes will be required.

Agriculture↗

Genetic and maternal environmental factors influencing litter size and reproductive efficiency in mice.

Lines of mice which have been genetically differentiated in litter size at birth and 6-week body weight by single-trait or index selection were randomly assigned to be reared in postnatal litter sizes (PLS) of N8, N12 or N16. The lines were selected as follows: large litter size (L+), large 6-week body weight (W+), selection index to decrease litter size and increase 6-week body weight (L-W+), selection index to increase litter size and decrease 6-week weight (L+W-) and random selection (K). Line and PLS differences in female body weight at weaning and 6 weeks were significant (P less than .05), with W+ greater than L+ greater than L-W+ greater than K greater than L+W- and N8 greater than N12 greaater than N16. Although there was a significant interaction between line and PLS, ranking of the lines was the same at each PLS. Females of each line by PLS group were exposed to males of the same group. Line and PLS differences (P less than .05) in female body weight at copulatory plug, adjusted for age, exhibited the same rankings as the earlier weights. Litter size at birth (dead plus alive), number born alive and number born alive per female exposed each revealed similar differences (P less than .05) in the ranking of lines and PLS. No significant interactions were found. Means for litter size were as follows for each line: L+ (17.4) greater than [W+ (14.1), L+W- (14.3)] greater than K (12.0) greater than L-W+ (10.8), and for each PLS: [N8 (14.0), N12 (13.9)] greater than N16 (13.2). The decline in litter size as PLS increased did not differ significantly from linearity. Adjusting litter size for body weight at copulatory plug eliminated the significant effect of PLS [N8 (13.9), N12 (13.7), N16 (13.4)]. The established genetic differences in litter size among lines due to selection were considerably larger than the maternal environmental effect due to postnatal litter size. Because of the negligible interaction between line and PLS, valid inferences concerning genetic differences among lines in litter size may be made at any postnatal litter size between eight and 16.

Animal Nutritional Physiological Phenomena↗

Perspectives for artificial insemination and genomics to improve global swine populations.

Civilizations throughout the world continue to depend on pig meat as an important food source. Approximately 40% of the red meat consumed annually worldwide (94 million metric tons) is pig meat. Pig numbers (940 million) and consumption have increased consistent with the increasing world population (FAO 2002). In the past 50 years, research guided genetic selection and nutrition programs have had a major impact on improving carcass composition and efficiency of production in swine. The use of artificial insemination (AI) in Europe has also had a major impact on pig improvement in the past 35 years and more recently in the USA. Several scientific advances in gamete physiology and/or manipulation have been successfully utilized while others are just beginning to be applied at the production level. Semen extenders that permit the use of fresh semen for more than 5 days post-collection are largely responsible for the success of AI in pigs worldwide. Transfer of the best genetics has been enabled by use of AI with fresh semen, and to some extent, by use of AI with frozen semen over the past 25 years. Sexed semen, now a reality, has the potential for increasing the rate of genetic progress in AI programs when used in conjunction with newly developed low sperm number insemination technology. Embryo cryopreservation provides opportunities for international transport of maternal germplasm worldwide; non-surgical transfer of viable embryos in practice is nearing reality. While production of transgenic animals has been successful, the low level of efficiency in producing these animals and lack of information on multigene interactions limit the use of the technology in applied production systems. Technologies based on research in functional genomics, proteomics and cloning have significant potential, but considerable research effort will be required before they can be utilized for AI in pig production. In the past 15 years, there has been a coordinated worldwide scientific effort to develop the genetic linkage map of the pig with the goal of identifying pigs with genetic alleles that result in improved growth rate, carcass quality, and reproductive performance. Molecular genetic tests have been developed to select pigs with improved traits such as removal of the porcine stress (RYR1) syndrome, and selection for specific estrogen receptor (ESR) alleles. Less progress has been made in developing routine tests related to diseases. Major research in genomics is being pursued to improve the efficiency of selection for healthier pigs with disease resistance properties. The sequencing of the genome of the pig to identify new genes and unique regulatory elements holds great promise to provide new information that can be used in pig production. AI, in vitro embryo production and embryo transfer will be the preferred means of implementing these new technologies to enhance efficiency of pig production in the future.

Animals↗

Histochemical studies on genetical control of hormonal enzyme inducibility in the mouse. V. Histochemical evidence for androgen inducibility of beta-glucuronidase in the epididymis.

The enzyme beta-glucuronidase (beta G) is shown, by histochemical methods, to be androgen inducible in the mouse epididymis. This trait has previously been believed to exist only in the kidney. Its presence in the genital tract constitutes a valuable tool in study of the developmental genetics of the reproductive system. Data presented here and previously imply co-ordinated genetic control of heterogeneous lysosomal populations. The results reported also provide a system for study of X-chromosomal activation, and of the developmental androgen dependence of the epididymis.

Androgens↗

Calcium homeostasis and human evolution.

The maintenance of calcium homeostasis is critical to survival. The systems dependent upon the availability of calcium include the musculoskeletal, nervous, and immune systems, all of which utilize the ionic properties of divalent calcium to perform critical functions. The widespread utilization of calcium in essential biological processes across a wide range of phyla argues strongly for its adoption early in the evolution of life. In the broad context of evolution, the use of calcium in the structure of the skeleton is a relatively late, or derived trait. It is not surprising, therefore, that when conditions demand the resorption of bone to sustain normal blood calcium levels, resorption can lead to serious impairment of function. Nowhere is the force of natural selection greater than when traits essential to successful reproduction are directly involved. Thus, when the requirements of pregnancy and lactation demand it, major shifts in the endocrine regulation of calcium turnover occur. While beneficial to reproductive success, elements of these changes can have deleterious effects later in life. However, selective pressure on such traits diminishes with the years of remaining reproductive life. The genetic determinants of traits that only manifest their deleterious aspects after reproduction has ceased altogether are thus able to remain and even increase in frequency in the population's gene pool. Such traits have been designated antagonistic pleiotropies. The following discussion explores the possibilities of Type I osteoporosis, a condition experienced by many postmenopausal women being an example of antagonistic pleiotropy.

Aged↗

A genetic evaluation of mating system and determinants of individual reproductive success in Atlantic salmon (Salmo salar L.).

The primary objective of this study was to use highly polymorphic microsatellite loci to estimate individual reproductive success in Atlantic salmon based on the number of surviving juveniles (young of the year) at the population level under natural conditions. We inferred reproductive strategies adopted by both sexes by applying a maximum likelihood method to determine parent-offspring genotype relationships. A high degree of variance in individual reproductive success for both males and females was revealed. The high number of mates used by both sexes is not concordant with previous behavioral studies proposing that females are mainly monogamous in this species. We found little evidence supporting the prediction from previous reports of a positive relationship between individual size and realized reproductive success for either males or females. For both sexes, however, there was a significant correlation between the number of mates and the number of offspring. These results indicate that this species' mating system is more flexible than previously thought and suggest that factors such as potential genetic benefits or environmental uncertainty may also be driving the evolution and the plasticity of mating systems in Atlantic salmon.

Alleles↗

Disassociation between weak sexual isolation and genetic divergence in a hermaphroditic land snail and implications about chirality.

Examination of the association between reproductive isolation and genetic divergence in a variety of organisms is essential for elucidating the mechanisms causing speciation. However, such studies are lacking for hermaphrodites. We measured premating (sexual) isolation in species pairs of the hermaphroditic land snail Albinaria and we compared it with their genetic divergence. We did not find substantial sexual isolation barriers between the species studied. The absence of strong sexual isolation between species implies its minor effect in the evolution of this genus, because distributional, population and life-history characteristics of Albinaria make mate-choice possibly redundant. Furthermore, we found disassociation between genetic divergence and sexual isolation, suggesting that they do not form necessarily a cause-effect duet. However, Albinaria voithii, the only dextral Albinaria species, shows strong sexual isolation against the other sinistral species. We discuss whether change in coiling either has triggered instantaneous speciation, or is an example of character displacement.

Animals↗

Divergent selection for mature body weight in dwarf White Leghorns. 1. Growth and reproductive responses to selection.

Genetic selection for high (H) and low (L) 20-week body weight (BW) in dwarf White Leghorns was practiced for three generations. The base population was the Oregon State University dwarf Leghorn population (C) selected previously for part year hen-housed egg production (EP), early sexual maturity (SM), and large initial egg weight (EW). Asymmetry of response was observed in both sexes. Realized heritabilities for 20-week BW for H and L males were .49 and .36 and for H and L females were .45 and .79, respectively. Twenty-week BW of the 3rd generation H and L line females were 1.48 and .80 kg, respectively, compared to 1.13 kg for the C line. Similar weights for the males were 1.74, .97, and 1.27 kg for the H, L, and C line, respectively. The H and L lines diverged significantly in 10, 20, 40, and 60-week BW, SM, and EW after one generation of selection and the divergence continued through the third generation. The EP and egg specific gravity did not differ among lines. The C line was intermediate to the selected lines for all traits except EP. The EW at 35 and 60 weeks differed between the H and L lines by 10 g at generation 3 and SM was 149 and 157 days for the H and L lines, respectively.

Animals↗

Professional self-regulation for preimplantation genetic diagnosis: experience of the American Society for Reproductive Medicine and other professional societies.

Professional societies that are stakeholders in preimplantation genetic diagnosis have democratically elected officers and favor self-regulation. Their practice guidelines are cognizant of societal needs and allow for introduction of new technologies. These societies have demonstrated the ability for professional self-regulation by collection, validation, and promulgation of center-specific results.

Fertilization in Vitro↗

Sexual conflict selects for male and female reproductive characters.

BACKGROUND: Strict genetic monogamy leads to sexual harmony because any trait that decreases the fitness of one sex also decreases the fitness of the other. Any deviation from monogamy increases the potential for sexual conflict. Conflict is further enhanced by sperm competition, and given the ubiquity of this phenomenon, sexual conflict is rife. In support of theory, experimentally enforced monogamy leads to the evolution of sexual benevolence. In contrast, with multiple mating, males evolve traits causing massive female fitness reductions when female evolution is restrained. Theory also predicts increased investment in spermatogenesis when sperm competition risk is high. While this supposition has correlational support, cause and effect has yet to be firmly established. RESULTS: By enforcing monogamy or polyandry in yellow-dung-fly lines, we have shown experimentally that males from polyandrous treatments evolved larger testes. Furthermore, females from this treatment evolved larger accessory sex glands. These glands produce a spermicidal secretion, so larger glands could increase female ability to influence paternity. Using molecular techniques, we have shown that, consistent with this idea, males' success as second mates is reduced in females from the polyandrous treatment. Nevertheless, males from polyandrous lines achieve higher paternity during sperm competition, and this finding further supports the testis evolution patterns. CONCLUSIONS: These results provide direct experimental support for macroevolutionary patterns of testis size evolution. Furthermore, we have shown that sperm competition selects for traits likely to be important in sexual conflicts over paternity, a result only previously demonstrated in Drosophila melanogaster.

Animals↗

Genetic counselling: ethical mediation of eugenic futures?

This paper examines some proposals concerning the involvement of genetic counsellors in reproductive decisionmaking. This involvement represents the future situation as being shaped decisively by the "relevant" knowledge of powerful medical and other political interests. The paper deconstructs and reorders this proposed knowledge in order to make it problematic. The projected involvement of genetic counselling backed by claims of ethical expertise is rendered denaturalised and particular, opening a conceptual space for the emergence of other futures. An alternative future in which public communication, not private medical decisions, is given as the primary ethical focus.

Abortion, Eugenic↗

Attitudes toward genetic testing and fetal diagnosis, 1990-1996.

With few exceptions, existing research on attitudes toward genetic testing and prenatal diagnosis is based on small studies using nonprobability samples of specialized populations. In this paper, we use a nationally representative sample from the General Social Survey to report on attitudinal change between 1990 and 1996, and to explore socio-demographic predictors of public views on genetic technology and reproduction in the context of changing mass media coverage between 1988 and 1995. During that period, media coverage of prenatal testing became both less frequent and less favorable, despite increasing use of this technology, whereas media reports about other types of genetic testing increased in frequency and became more favorable. Between 1990 and 1996, attitudes toward genetic testing remained stable, although the attitudes of specific demographic subgroups may be changing in different directions. Attitudes toward abortion in case of genetic defect became more negative over the period studied. We explore some of the implications of these findings.

Abortion, Induced↗

Reproductive performance of chimeric mice produced from genetic lines that differ in litter size.

A population of chimeras was made by aggregating 8- and 16-cell embryos from two mouse strains: a randomly bred line (C) and a selected line characterized by large litters (JU), with litter sizes of 7.7 and 13.5, respectively. The two genotypes were developmentally "balanced", as judged by the high frequency (90%) of chimeras with an intermediate or high degree of coat-color chimerism, a chimeric sex ratio of 2.2:1 males:females, and a high percentage of chimeras (31% of males, 71% of females) with germ cells of both strains. Litter size characteristics, including ovulation rate, implantation rate, rates of pre- and postimplantation embryo survival and number born were studied in the female chimeras and compared with the performance of both parent lines and to the genetic cross of the two lines. Values for JU females exceeded those for C females for all parameters studied except postimplantation embryo survival, which was the same for both lines in second litters and was lower for JU's third litters. For most traits, means for genetic crossbreds and chimeras were similar, regardless of whether the means were at or above the midparent average. In contrast, for ovulation rate and body weight, genetic crossbreds and chimeras clearly differed, with chimeric females being similar to the JU line and genetic crossbred females exhibiting additive inheritance. Because of phenotypic differences between experimental chimeras and crossbreds produced from the same two lines, chimeras may provide a useful model for studying the physiologic basis for expression of genetic differences in quantitative traits.

Animals↗

The genetics of species differences.

Species are separated by reproductive isolation as well as by more 'ordinary' differences in morphology and behavior that play no necessary role in blocking gene flow. Although a great deal is now known about the genetics of reproductive isolation, we are only beginning to understand the genetic basis of ordinary phenotypic differences between species. I review what is known about the number of genes involved in such differences, as well as about the role of major genes and epistasis in the evolution of these differences. I also compare and contrast these findings with those on the genetics of reproductive isolation.

Journal Article↗

Male reproductive timing in Rhesus macaques is influenced by the 5HTTLPR promoter polymorphism of the serotonin transporter gene.

The 5HTTLPR polymorphism in the promoter region of the human serotonin transporter (SLC6A4) gene is known to be associated with various stress-related psychological and psychiatric phenomena. We observed that a similar diallelic polymorphism in the orthologous gene of rhesus macaques (Macaca mulatta) was related to the reproductive life history of 580 males residing in the free-ranging colony of Cayo Santiago, Puerto Rico, between 1985 and 1998. At first glance, the polymorphism appeared to be selectively neutral because no difference in total reproductive output was noted between males of different 5HTTLPR genotypes. However, whereas heterozygotes were significantly more reproductive than homozygotes at intermediate age (10-13 yr), the opposite held true before and after this period (n = 682 offspring; randomization P = 0.014). This association, which explains approximately 7% of the observed variation in sire age, most likely reflects different natal dispersal patterns and represents the first reported instance of a genetic influence on reproductive timing in mammals.

Age Factors↗

Joint analysis of the influence of CYP11B1 and DGAT1 genetic variation on milk production, somatic cell score, conformation, reproduction, and productive lifespan in German Holstein cattle.

Recent publications indicate genetic variation in milk production traits on proximal BTA14, which cannot be explained solely with genetic variation in the DGAT1 gene. To elucidate these QTL effects, animals from a German Holstein granddaughter design (18 families, 1,291 sons) were genotyped for CYP11B1 (V30A) and DGAT1 (K232A) polymorphisms. Frequencies of alleles of maternal descent were estimated for CYP11B1(V) (0.776) and DGAT1(K) (0.549). Allele substitution effects (alpha/2) were first calculated for both alleles in separate models and then in a joint model. From the joint analysis, CYP11B1(V) effects on fat content (+0.04%) and protein content (+0.01%) were positive. Effects on milk yield (-82 kg), fat yield (-0.5 kg), and protein yield (-1.9 kg) were negative. Compared with the individual analysis, DGAT1(K) effects on fat content (+0.28%), protein content (+0.06%), and milk yield (-258 kg) were reduced; fat yield (+10.8 kg) was enhanced; and protein yield (-3.8 kg) was reduced. In the joint analysis, allele substitution effects of CYP11B1(V) and DGAT1(K) together explained more of the variation in milk production traits than DGAT1(K) alone. Further significant effects were found for CYP11B1(V) and DGAT1(K) among 6 reproduction traits and 14 conformational traits. These observations indicate a possible negative influence of DGAT1(K) on maternal nonreturn rate, and thus, on length of productive life.

Animals↗

Speciation by postzygotic isolation: forces, genes and molecules.

New species arise as reproductive isolation evolves between diverging populations. Here we review recent work in the genetics of postzygotic reproductive isolation-the sterility and inviability of species hybrids. Over the last few years, research has taken two new directions. First, we have begun to learn a good deal about the population genetic forces driving the evolution of postzygotic isolation. It has, for instance, become increasingly clear that conflict-driven processes, like sexual selection and meiotic drive, may contribute to the evolution of hybrid sterility. Second, we have begun to learn something about the identity and molecular characteristics of the actual genes causing hybrid problems. Although molecular genetic data are limited, early findings suggest that "speciation genes" correspond to loci having normal functions within species and that these loci sometimes diverge as a consequence of evolution in gene regulation.

Animals↗

Restriction fragment length polymorphism analysis of open reading frame 5 gene of porcine reproductive and respiratory syndrome virus isolates in Korea.

The genetic variability of porcine reproductive and respiratory syndrome virus (PRRSV) was studied by restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR)-amplified fragments among 50 Korean isolates from open reading frame 5. All Korean PRRSVs were isolated from the field cases after the marketing of an U.S. ATCC VR2332-derived modified live PRRSV vaccine. Combining the restriction enzyme digestion patterns obtained with MluI, HincII, SacII, and HaeIII, we observed 19 distinct RFLP patterns. Seventeen out of 50 PRRSV isolates (34%) exhibited the modified live PRRSV vaccine RFLP pattern. The genomic variations that have been identified in the present study seemed to represent characteristic features of the Korean PRRSV isolates. PCR-based RFLP analysis using several restriction enzymes provides a good genetic estimate for isolate differentiation.

Animals↗