Significance of specific-locus germ-cell mutations in mice

On the basis of a historical-control incidence of 39 mutants in 688 921 progeny (5.7 mutants /10⁵ animals or 0.82 mutations/locus/10⁵ gametes) proposals are made for the numbers of test progeny required when screening for possible mutagens using the specific-locus test in mice (7 loci).It is recommended that 25 000 control progeny should be included in each test, to establish homogeneity with the historical controls. This would also be the number of progeny required from treated males, unless significantly positive results had been obtained with smaller numbers.It would appear that the greater sensitivity of post-spermatogonial stages could more than compensate for practical difficulties in sampling these stages, rather than spermatogonia, in screening tests.     

Tests for urethane induction of germ-cell mutations and germ-cell killing in the mouse

Urethane, a chemical that has given varied results in mutagenesis assays, was tested in the mouse specific-locus test, and its effect on germ-cell survival was explored. Altogether 32,828 offspring were observed from successive weekly matings of males exposed to the maximum tolerated i.p. dose of 1750 mg urethane/kg. The combined data rule out (at the 5% significance level) an induced mutation rate greater than 1.7 times the historical control rate. For spermatogonial stem cells alone, the multiple ruled out is 3.2, and for poststem-cell stages, 3.5. Litter sizes from successive conceptions made in any of the first 7 weeks give no indication of induced dominant lethality, confirming results of past dominant-lethal assays. That urethane (or an active metabolite) reaches germ cells is indicated by SCE induction in spermatogonia demonstrated by other investigators. Cytotoxic effects in spermatogonia are suggested by our finding of a slight reduction in numbers of certain types of spermatogonia in seminiferous tubule cross-sections and of a borderline decrease in the number of litters conceived during the 8th and 9th posttreatment weeks. The negative results for induction of gene mutations as well as clastogenic damage are at variance with Nomura's reports of dominant effects (F1 cancers and malformations) produced by urethane.     

Significance of specific-locus germ-cell mutations in mice.

On the basis of a historical-control incidence of 39 mutants in 688 921 progeny (5.7 mutants/10(5) animals or 0.82 mutations/locus/10(5) gametes) proposals are made for the numbers of test progeny required when screening for possible mutagens using the specific-locus test in mice (7 loci). It is recommended that 25,000 control progeny should be included in each test, to establish homogeneity with the historical controls. This would also be the number of progeny required from treated males, unless significantly positive results had been obtained with smaller numbers. It would appear that the greater sensitivity of post-spermatogonial stages could more than compensate for practical difficulties in sampling these stages, rather than spermatogonia, in screening tests.     

Allometric estimation of metabolic rates in animals

The relationship between body mass (M) and metabolic rate (MR) typically accounts for most (>90%) of the inter-specific variation in MR. As such, when measurement of a species of interest is not possible, its MR can often be predicted using M. However, choosing an appropriate relationship to make such predictions is critical, and the choice is complicated by ongoing debate about the structure of the relationship between M and MR. The present study examines a range of methods including ordinary least squares (OLS), reduced major axis (RMA), and phylogenetically-informed (PI) approaches for estimating log(MR) from log(M), as well as non-linear approaches for estimating the relationship between MR and M without the need for log-transformation. Using data for the basal metabolic rates of mammals, it is shown that RMA regression overestimates the scaling exponent of MR (b, where MR=aM(b)), suggesting that OLS regression is appropriate for these data. PI approaches are preferred over non-PI ones, and the best estimates of log(MR) are obtained by including information on body temperature, climate, habitat, island endemism, and use of torpor in addition to log(M). However, the use of log-transformed data introduces bias into estimates of MR, while the use of non-linear regression underestimates MR for small mammals. This suggests that no single relationship is appropriate for describing the relationship between MR and M for all mammals, and that relationships for more narrow taxonomic groups or body mass ranges should be used when predicting MR from M.     

鱼类生殖细胞移植的研究进展

生殖细胞移植在生物学、畜牧学及兴起的鱼类生物工程上都有很多用途。目前,采用基于基因组的育种方法已选育出多种带有所需遗传性状的鱼类新品系;但由于缺乏鱼卵与胚胎的低温保存技术,尚未找到长期保存其种质资源的方法。就鱼类生殖细胞移植的最新研究进展进行综述,同时针对鱼类生殖细胞移植及其低温保存技术研究中存在的问题进行讨论,并提出了鱼类生殖细胞移植的应用前景,以期能积极推动鱼类生物工程的研究,加速鱼类生殖细胞移植在产业中的应用。     

Interphase cytogenetics in estimation of genomic mutations in somatic cells

The review considers the current state, possibilities, and perspectives of using interphase cytogenetics in the estimation of genomic mutations in human and animal somatic cells for aims of genetic toxicology and genetic instability analysis. Possible mechanisms underlying action of mutagens causing numeric chromosome aberrations are discussed.     

Interphase cytogenetics in estimation of genomic mutations in somatic cells

The review considers the current state, possibilities, and perspectives of using interphase cytogenetics in the estimation of genomic mutations in human and animal somatic cells for aims of genetic toxicology and genetic instability analysis. Possible mechanisms underlying action of mutagens causing numeric chromosome aberrations are discussed.Translated from Genetika, Vol. 41, No. 1, 2005, pp. 5–16.Original Russian Text Copyright © 2005 by Timoshevsky, Nazarenko.     

Genomic analysis of gamma-ray-induced germ-cell mutations at the b locus recovered from the medaka specific-locus test.

To study how gamma-ray-induced germ-cell mutations are fixed at the early embryonic stage of the next generation, genomic alterations in the b locus mutants (colorless melanophores) detected during development in the medaka specific-locus test (SLT) were analyzed. First, nine anonymous DNA markers linked to the b locus were cloned and mapped into the region extending about 47cM surrounding the b locus. Next, losses of paternal alleles of these DNA markers were examined in each of the 51 gamma-ray-induced b locus mutants obtained after irradiation of sperm or spermatids. In these mutants, 47 were dominant lethals, three were semi-viable and one was viable. All the mutants examined had large deletions surrounding the b locus. One viable mutant had an interstitial deletion, while all the semi-viable and dominant lethal ones appeared to have terminal deletions. Deletions extending about 20-35cM were the most frequently observed in 18 of the 51 mutants examined. The largest one extended more than 40cM. These results suggest that most of the gamma-ray induced germ cell mutations recovered as total specific-locus mutants were accompanied by large genomic deletions, which eventually led the mutant embryos to dominant lethality.     

Abundance estimation of long-diving animals using line transect methods

Line transect sampling is one of the most widely used methods for estimating the size of wild animal populations. An assumption in standard line transect sampling is that all the animals on the trackline are detected without fail. This assumption tends to be violated for marine mammals with surfacing/diving behaviors. The detection probability on the trackline is estimated using duplicate sightings from double-platform line transect methods. The double-platform methods, however, are insufficient to estimate the abundance of long-diving animals because these animals can be completely missed while the observers pass. We developed a more flexible hazard probability model that incorporates information on surfacing/diving patterns obtained from telemetry data. The model is based on a stochastic point process and is statistically tractable. A simulation study showed that the new model provides near-unbiased abundance estimates, whereas the traditional hazard rate and hazard probability models produce considerably biased estimates. As an illustration, we applied the model to data on the Baird's beaked whale (Berardius bairdii) in the western North Pacific.     

Somatic and germ-cell sex in mammals

The phenotypic sex of an individual mammal is determined by the sex of its gonads, i.e. testes or ovaries. This in turn is determined by the presence or absence of a small region of the Y chromosome, located near the X-Y pairing region in man and on the short arm of the Y chromosome in the mouse. The testis-determining region of the Y appears to exert its primary effect by directing the supporting-cell lineage of the gonad to differentiate as Sertoli cells, acting at least in part cell-autonomously. The phenotypic sex of a germ cell, i.e. whether it undergoes spermatogenesis or oogenesis, is determined at least in the mouse by whether or not it enters meiotic prophase before birth. This depends not on its own sex chromosome constitution, but on its cellular environment. A germ cell in or near normal testis cords (made up mainly of Sertoli cells) is inhibited from entering meiosis until after birth; one that escapes this inhibition will develop into an oocyte even if it is in a male animal and is itself XY in chromosome constitution.     

Long-distance photogrammetric trait estimation in free-ranging animals: A new approach

The acquisition of accurate information on the size of traits in animals is fundamental for the study of animal ecology and evolution and their management. We demonstrate how morphological traits of free-ranging animals can reliably be estimated on very large observation distances of several hundred meters by the use of ordinary digital photographic equipment and simple photogrammetric software. In our study, we estimated the length of horn annuli in free-ranging male Alpine ibex (Capra ibex) by taking already measured horn annuli of conspecifics on the same photographs as scaling units. Comparisons with hand-measured horn annuli lengths and repeatability analyses revealed a high accuracy of the photogrammetric estimates. If length estimations of specific horn annuli are based on multiple photographs measurement errors of <5.5 mm can be expected. In the current study the application of the described photogrammetric procedure increased the sample size of animals with known horn annuli length by an additional 104%. The presented photogrammetric procedure is of broad applicability and represents an easy, robust and cost-efficient method for the measuring of individuals in populations where animals are hard to capture or to approach.     

Mechanisms of germ-cell specification in mouse embryos

The mode and timing of germ-cell specification has been studied in diverse organisms, however, the molecular mechanism regulating germ-cell-fate determination remains to be elucidated. In some model organisms, maternal germ-cell determinants play a key role. In mouse embryos, some germ-line-specific gene products exist as maternal molecules and play critical roles in a pluripotential cell population at preimplantation stages. From those cells, primordial germ cells (PGCs) are specified by extracellular signaling mediated by tissue, as well as cell-cell interaction during gastrulation. Thus, establishment of germ-cell lineage in mammalian embryos appears to be regulated by a multistep process, including formation and maintenance of a pluripotential cell population, as well as specification of PGCs. PGCs can be generated from pluripotential embryonic stem (ES) cells in a simple monolayer culture in which tissue interaction does not occur. This raises the possibility that ES cells, as well as, possibly, pluripotential cells in preimplantation embryos, are more closely related to the PGC precursors than pluripotential cells after implantation.     

Muscle parameters estimation based on biplanar radiography

The evaluation of muscle and joint forces in vivo is still a challenge. Musculo-Skeletal (musculo-skeletal) models are used to compute forces based on movement analysis. Most of them are built from a scaled-generic model based on cadaver measurements, which provides a low level of personalization, or from Magnetic Resonance Images, which provide a personalized model in lying position. This study proposed an original two steps method to access a subject-specific musculo-skeletal model in 30 min, which is based solely on biplanar X-Rays. First, the subject-specific 3D geometry of bones and skin envelopes were reconstructed from biplanar X-Rays radiography. Then, 2200 corresponding control points were identified between a reference model and the subject-specific X-Rays model. Finally, the shape of 21 lower limb muscles was estimated using a non-linear transformation between the control points in order to fit the muscle shape of the reference model to the X-Rays model. Twelfth musculo-skeletal models were reconstructed and compared to their reference. The muscle volume was not accurately estimated with a standard deviation (SD) ranging from 10 to 68%. However, this method provided an accurate estimation the muscle line of action with a SD of the length difference lower than 2% and a positioning error lower than 20 mm. The moment arm was also well estimated with SD lower than 15% for most muscle, which was significantly better than scaled-generic model for most muscle. This method open the way to a quick modeling method for gait analysis based on biplanar radiography.     

Prediction of haplotypes for ungenotyped animals and its effect on marker-assisted breeding value estimation

Background

In livestock populations, missing genotypes on a large proportion of animals are a major problem to implement the estimation of marker-assisted breeding values using haplotypes. The objective of this article is to develop a method to predict haplotypes of animals that are not genotyped using mixed model equations and to investigate the effect of using these predicted haplotypes on the accuracy of marker-assisted breeding value estimation.

Methods

For genotyped animals, haplotypes were determined and for each animal the number of haplotype copies (nhc) was counted, i.e. 0, 1 or 2 copies. In a mixed model framework, nhc for each haplotype were predicted for ungenotyped animals as well as for genotyped animals using the additive genetic relationship matrix. The heritability of nhc was assumed to be 0.99, allowing for minor genotyping and haplotyping errors. The predicted nhc were subsequently used in marker-assisted breeding value estimation by applying random regression on these covariables. To evaluate the method, a population was simulated with one additive QTL and an additive polygenic genetic effect. The QTL was located in the middle of a haplotype based on SNP-markers.

Results

The accuracy of predicted haplotype copies for ungenotyped animals ranged between 0.59 and 0.64 depending on haplotype length. Because powerful BLUP-software was used, the method was computationally very efficient. The accuracy of total EBV increased for genotyped animals when marker-assisted breeding value estimation was compared with conventional breeding value estimation, but for ungenotyped animals the increase was marginal unless the heritability was smaller than 0.1. Haplotypes based on four markers yielded the highest accuracies and when only the nearest left marker was used, it yielded the lowest accuracy. The accuracy increased with increasing marker density. Accuracy of the total EBV approached that of gene-assisted BLUP when 4-marker haplotypes were used with a distance of 0.1 cM between the markers.

Conclusions

The proposed method is computationally very efficient and suitable for marker-assisted breeding value estimation in large livestock populations including effects of a number of known QTL. Marker-assisted breeding value estimation using predicted haplotypes increases accuracy especially for traits with low heritability.     

Risk assessment of meat and milk from cloned animals

Research on, and commercialization of, cloned cattle has been conducted for more than 20 years. Early techniques relied on the physical splitting of embryos or using embryo cells for nuclear transfer to generate cloned animals. Milk and meat from these animals entered into the human food market with no evidence of problems. With the advent of nuclear transfer, which enables the direct transference and preservation of high-value meat- and milk-producing genotypes to offspring, concerns have been raised about whether the products from somatic cell nuclear transfer-produced animals are safe for human consumption. Studies on the biochemical properties of food products from cloned and noncloned animals have thus far not detected any differences. All data to date indicate no significant differences in the measured parameters between animals created by nuclear transfer and normally bred animals. Public acceptance of cloned animal products depends upon forthcoming US Food and Drug Administration approval along with convincing safety data.     

Abundance estimation of diving animals by the double-platform line transect method

In conventional line transect theory, it is assumed that all animals on the line are detected. This article introduces an extended and generalized hazard probability model without the need for such an assumption. The proposed method needs a survey design with independent observers having the same visual region and assumes an explicit distinction of simultaneous and delayed duplicates. It can take account of random heterogeneity caused by surfacing behavior as well as systematic heterogeneity by covariate effects. Furthermore, it can be easily extended to cases in which data from incompletely independent observers are available. The abundance estimate is based on the Horvitz-Thompson estimator in unequal detectability sampling scheme. Simulation studies suggest that the proposed method has good performance. The method is applied to a real data set on Antarctic minke whales in the illustration.     

Germ-plasm formation and germ-cell determination in Drosophila.

Germ cells in Drosophila form in a specialized cytoplasm that is critical for germ-cell development. Several genes involved in this process have been identified. By studying the temporal and functional sequence of interactions between their products, these genes can be ordered into a pathway for germ-plasm formation.