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PubMed1990

Stochastic modeling of multiple ovulation and embryo transfer breeding schemes in small closed dairy cattle populations.

Genetic changes and genetic drift in three small closed dairy cattle populations were examined with a stochastic simulation model. Multiple ovulation and embryo transfer and AI techniques were simulated in three populations, two with 88 breeding females each and one with 352 breeding females. The selection goal was to maximize genetic improvement in milk yield. The reduction in genetic variation due to inbreeding and linkage disequilibrium was accounted for in the simulation. Strict restriction against inbred mating slowed genetic progress significantly in the small population but would not be consequential in the larger population. However, allowing inbred mating in the smaller population caused a rapid accumulation of inbreeding. Linkage disequilibrium was as important as inbreeding in reducing genetic variation. Genetic drift variance was much smaller in the larger population.

Animals↗
PubMed2006

Computational models for neurogenic gene expression in the Drosophila embryo.

The early Drosophila embryo is emerging as a premiere model system for the computational analysis of gene regulation in development because most of the genes, and many of the associated regulatory DNAs, that control segmentation and gastrulation are known. The comprehensive elucidation of Drosophila gene networks provides an unprecedented opportunity to apply quantitative models to metazoan enhancers that govern complex patterns of gene expression during development. Models based on the fractional occupancy of defined DNA binding sites have been used to describe the regulation of the lac operon in E. coli and the lysis/lysogeny switch of phage lambda. Here, we apply similar models to enhancers regulated by the Dorsal gradient in the ventral neurogenic ectoderm (vNE) of the early Drosophila embryo. Quantitative models based on the fractional occupancy of Dorsal, Twist, and Snail binding sites raise the possibility that cooperative interactions among these regulatory proteins mediate subtle differences in the vNE expression patterns. Variations in cooperativity may be attributed to differences in the detailed linkage of Dorsal, Twist, and Snail binding sites in vNE enhancers. We propose that binding site occupancy is the key rate-limiting step for establishing localized patterns of gene expression in the early Drosophila embryo.

Animals↗
PubMed2006

The developmental neurotoxicity of fipronil: notochord degeneration and locomotor defects in zebrafish embryos and larvae.

Fipronil is a phenylpyrazole insecticide designed to selectively inhibit insect gamma-aminobutyric acid (GABA) receptors. Although fipronil is often used in or near aquatic environments, few studies have assessed the effects of this neurotoxicant on aquatic vertebrates at sensitive life stages. We explored the toxicological effects of fipronil on embryos and larvae using the zebrafish (Danio rerio) experimental model system. Embryos exposed to fipronil at nominal concentrations at or above 0.7 microM (333 mug/l) displayed notochord degeneration, shortening along the rostral-caudal body axis, and ineffective tail flips and uncoordinated muscle contractions along the body axis in response to touch. This phenotype closely resembles zebrafish locomotor mutants of the accordion class and is consistent with loss of reciprocal inhibitory neurotransmission by glycinergic commissural interneurons in the spinal cord. Consistent with the hypothesis that notochord degeneration may be due to abnormal mechanical stress from muscle tetany, the expression patterns of gene and protein markers specific to notochord development were unaffected by fipronil. Moreover, the degenerative effects of fipronil (1.1 microM) were reversed by coexposure to the sodium channel blocker MS-222 (0.6mM). The notochord effects of fipronil were phenocopied by exposure to 70 microM strychnine, a glycinergic receptor antagonist. In contrast, exposure to gabazine, a potent vertebrate GABA(A) antagonist, resulted in a hyperactive touch response but did not cause notochord degeneration. Although specifically developed to target insect GABA receptors with low vertebrate toxicity, our results suggest that fipronil impairs the development of spinal locomotor pathways in fish by inhibiting a structurally related glycine receptor subtype. This represents an unanticipated and potentially novel mechanism for fipronil toxicity in vertebrates.

Animals↗
PubMedDate not supplied

The effects of nest temperature, nest substrate, and clutch size on the oxygenation of embryos and larvae of the Australian moss frog, Bryobatrachus nimbus.

The jelly around amphibian eggs presents a formidable barrier to oxygen diffusion. Therefore, egg capsules must be thin enough, and the dimensions of globular egg masses small enough, to avoid oxygen limitation leading to developmental retardation or death. The eggs of the Australian moss frog, Bryobatrachus nimbus, have the thickest jelly capsule known for any anuran amphibian. Laboratory measurements of respirometric variables predict that single prehatching embryos should be normoxic between 5 degrees and 20 degrees C, with Po(2 in) maintained above critical levels (10.2-17.0 kPa). However, numerical models of embryos amid larger egg masses (13-20 eggs) predict hypoxia at temperatures above 5 degrees C. Contrary to model predictions, however, B. nimbus embryos rarely experience hypoxia in natural nests, because embryos occur in one or two layers and the moss substrate permits aeration of the lower surface while photosynthesis probably supplies oxygen directly. After hatching, larvae move to oxygen-rich regions of the jelly mass and disperse more widely within the mass as temperatures increase. Although nest characteristics relieve diffusive constraints, small clutch sizes, low rates of embryonic and larval respiration, and the cool climate occupied by B. nimbus are the main characteristics that prevent hypoxia.

Animals↗
PubMed2001

Neuroprotective effects of lamotrigine and remacemide on excitotoxicity induced by glutamate agonists in isolated chick retina.

The possible neuroprotective effects of two recently developed antiepileptic compounds, lamotrigine (LTG) and remacemide (REMA), against glutamate agonist-induced excitotoxicity have been investigated in the isolated chick embryo retina model. Retina segments from 15- or 16-day-old embryos were incubated in 1 ml of balanced salt solution, at 25 degrees C for 30 min, in the presence or absence of N-methyl-d-aspartate (NMDA), kainic acid (KA), or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) (10 to 200 microM). LTG, REMA, and the active desglycinyl metabolite of REMA (d-REMA) (10-200 microM) were added separately 5 min before glutamate agonists. Retina damage was assessed after 24 h (i) by measuring LDH activity present in the medium, expressed as percentage of total retina LDH activity, and (ii) by histological analysis of retina specimens through scoring for the presence or absence of edema, necrosis, nuclear pyknosis, and cell layer damage. LTG, REMA, and d-REMA reduced LDH release produced by NMDA 58-70% in a dose-dependent manner, with d-REMA being the most potent (EC(50): d-REMA, 25.75 +/- 3.27 microM; REMA, 64.75 +/- 7.75 microM; LTG, 60.50 +/- 6.80 microM; P < 0.001). The drugs had less effect on the LDH release produced by AMPA and KA. Histological analysis confirmed these biochemical results, with all three compounds reducing edema and the number of necrotic and pyknotic nuclei in the ganglion layer. d-REMA provided almost complete protection of the ganglion cell layer against damage produced by NMDA. Combinations of d-REMA and LTG showed additive rather than potentiative effects against NMDA-induced cell injury. The present data provide pharmacological evidence that LTG, REMA, and d-REMA decrease glutamate agonist-induced excitotoxicity in isolated chick retina, findings that might have therapeutic implications for various neurological disorders.

Acetamides↗
PubMed2004

The polychaete Platynereis dumerilii (Annelida): a laboratory animal with spiralian cleavage, lifelong segment proliferation and a mixed benthic/pelagic life cycle.

Platynereis dumerilii, a marine polychaetous annelid with indirect development, can be continuously bred in the laboratory. Here, we describe its spectacular reproduction and development and address a number of open research problems. Oogenesis is easily studied because the oocytes grow while floating in the coelom. Unlike the embryos of other model spiralians, the Platynereis embryo is transparent giving insight into the dynamic structures and processes inside the cells that accompany the prevailing anisotropic cleavages. Functional studies on cell specification and differential gene expression in embryos, larvae, and later stages are underway. Lifelong proliferation of uniform trunk segments qualifies Platynereis as a model for the study of gene expression and of the functional circuitry of this process. Platynereis can also become a stepping stone in the comparison of segmentation between annelids and arthropods because it comes closer to the putative ancestral morphology and style of development than other model annelids.

Animals↗
PubMed1992

Cardiac mechanics in the stage-16 chick embryo.

A theoretical model is presented for the tubular heart of the stage-16 chick embryo (2.3 days of a 21-day incubation period). The model is a thick-walled, pseudoelastic cylindrical shell composed of three isotropic layers: the endocardium, the cardiac jelly, and the myocardium. The analysis is based on a shell theory that accounts for large deformation, material nonlinearity, residual strain, and muscle activation, with material properties inferred from available experimental data. We also measured epicardial strains from recorded motions of microspheres on the primitive right ventricles of stage-16 white Leghorn chick embryos. Relative to end diastole, peak axial and circumferential Lagrange strains occurred near end systole and had similar values. The magnitudes of these strains varied along the longitudinal axis of the heart (-0.16 +/- 0.08), being larger near the ends of the primitive right ventricle and smaller near midventricle. The in-plane shear strain was less than 0.05. Comparison of theoretical and experimental strains during the cardiac cycle shows generally good agreement. In addition, the model gives strong stress concentrations in the myocardial layer at end systole.

Animals↗
PubMed2001

A two- versus three-embryo transfer: the oocyte donation model.

OBJECTIVE: To compare implantation and pregnancy rates in oocyte recipients undergoing a two-embryo versus three-embryo transfer, 3 days after retrieval. DESIGN: Retrospective comparative analysis. SETTING: University-based in vitro fertilization center. PATIENT(S): All oocyte recipients undergoing embryo transfer from January 1, 1997 through August 31, 1999. INTERVENTION(S): Recipients received two or three embryos. MAIN OUTCOME MEASURE(S): Implantation, and clinical and multiple pregnancy rates. RESULT(S): Seventy-three recipients underwent a two-embryo transfer, and 376 had three embryos replaced. The numbers of oocytes retrieved (12.7 +/- 0.89 vs. 13.1 +/- 0.36) and embryos obtained (8.05 +/- 0.65 vs. 8.77 +/- 0.27) did not differ between the two-embryo and three-embryo transfer groups, nor did the proportion of patients with embryo cryopreservation (54.3% vs. 42.6%, respectively). There was no significant difference in pregnancy or implantation rates when comparing those patients with a two-embryo transfer to those with a three-embryo transfer. Significantly, 13.8% of the pregnancies in the three-embryo transfer group were triplet. CONCLUSION(S): Reducing the number of embryos transferred in an oocyte donation cycle can lower the incidence of triplet pregnancies without significantly lowering the overall pregnancy rate.

Cryopreservation↗
PubMed1998

Amnio-allantoic fluid exchange for the prevention of intestinal damage in gastroschisis. III: Determination of the waste products removed by exchange.

Experimental amnio-allantoic fluid (AAF) exchange has been shown to prevent intestinal damage in the chicken embryo gastroschisis model. AAF contains both urinary and gastrointestinal waste products (UWP and GIWP). An experimental study was performed to find the waste products responsible for this intestinal damage. Gastroschisis was created in 20 chick embryos. Half were treated with AAF exchange, the other half were not treated. AAF samples were obtained for biochemical determination of urea nitrogen and creatinine as UWP, bile acids and bilirubin as GIWP at the end of the incubation. Intestines were evaluated by light microscopy. While GIWP (Bile salts and bilirubin) were significantly removed from AAF by exchange, the levels of UWP (urea nitrogen and creatinine) were unaffected. Intestinal wall thickness was less in the exchange group compared to the untreated group. The unchanged levels of UWP after AAF exchange may be attributed to their relatively rapid production compared to GIWP. Dilution of GIWPs by AAF exchange results in prevention of the intestinal damage in gastroschisis.

Allantois↗
PubMed2002

A health-economic decision-analytic model comparing double with single embryo transfer in IVF/ICSI.

BACKGROUND: Single embryo transfer (SET) is the sole strategy with which to reduce the incidence of twins following assisted reproductive technology (ART), but SET may increase the number of ART cycles needed per live-born child. Its cost-effectiveness compared with double embryo transfer (DET) is therefore unknown. METHODS: A decision-analytic model comparing SET with DET was developed. Estimates were obtained from literature, national pregnancy registers and local hospital records. A sensitivity analysis was performed, using pregnancy rates from four published studies. The outcome measure was the cost per child born, calculated from IVF procedure-related, pregnancy-related and neonatal care costs. Neonatal mortality and long-term morbidity costs were not taken into account. RESULTS: Independently of the pregnancy rates used, the SET cost per child born was in all instances the same as with DET, varying from EURO= 9520 (SET) versus EURO= 9511 (DET) to EURO= 12254 (SET) versus EURO= 12934 (DET). CONCLUSIONS: More ART cycles are required to obtain the same numbers of children born following SET compared with DET. Because SET allows the avoidance of twins and thus diminishes pregnancy-related and neonatal care costs, there is no difference in the cost per child born between SET and DET. The real advantage of SET is the avoidance of the very high long-term costs resulting from the increased morbidity of twins after birth.

Cost-Benefit Analysis↗
PubMed2005

New tools for visualization and analysis of morphogenesis in spherical embryos.

Many classical models of development, including amphibians, fish, and echinoderms, have embryos that are approximately spherical and contain concentric cell layers during early development. Neighbor relationships in such curved cell layers are not conveniently assayed or measured by conventional physical or optical sectioning techniques. To answer these challenges, we have constructed computational methods that correct for spherical distortion in 2D images and that allow extraction of concentric cell layers from 3D digital images. These methods for quantitative analysis and visualization of early development in spherical embryos are introduced by using them for the quantitative analysis of 2D and 3D images of gastrula stage Xenopus laevis.

Algorithms↗
PubMed1987

Arachidonic acid and male genital differentiation.

Arachidonic acid (AA) prevents neural tube/defects, cleft palate, and micrognathia in the rat models of diabetic embryopathy and neural tube defects in the mouse embryo culture model. In this study, the involvement of AA in the male genital differentiation was described. These observations raise the complementary possibility that the AA-prostaglandin biochemical pathway may be a major mechanism mediating many embryonic events that involve cellular movement and fusion.

Animals↗
PubMed1988

Rat whole embryo culture: an in vitro model for testing nitrous oxide teratogenicity.

The teratogenic effects of nitrous oxide (N2O) on postimplantation rat embryos were studied using a whole embryo culture system to separate the direct effects of N2O from those that are maternally mediated. A total of 100, 10-day-old rat embryos were cultured in either a control atmosphere (75% N2, 20% O2, and 5% CO2), or a N2O atmosphere (75% N2O, 20% O2, and 5% CO2). After 22 h of culture embryos were examined microscopically, and protein and DNA contents were determined. DNA content was significantly lower in the embryos exposed to N2O compared with the controls. Additionally, three malformed embryos and four embryos with left-sided tails were observed in the N2O group, whereas no abnormalities were observed in the control group. There were no differences in crown-rump length, somite numbers, limb bud index, and protein content between the two groups of embryos. The positive findings in this study indicate that whole embryo culture is useful for studying the mechanisms of N2O teratogenicity.

Abnormalities, Drug-Induced↗
PubMed1993

Restoration of normal implantation rates in mouse embryos with a hatching impairment by use of a new method of assisted hatching.

OBJECTIVE: To use a newly developed mouse model for embryos with a hatching defect to develop improved methods of assisted hatching. DESIGN: We had previously demonstrated that mouse embryos with reduced cell numbers, when resident within an intact zona, exhibit a hatching deficiency, and we used these "hatching-defective" embryos to demonstrate that hatching is mediated predominantly, if not entirely, by lysis of the zona pellucida. In the present study, the hatching-defective embryos were implanted into pseudo-pregnant females to determine if implantation rates were reduced. In addition, to compensate for reduced zona lysin production, the zonae of some embryos were circumferentially thinned with acid Tyrode's solution. The implantation rates and developmental potential of hatching-defective embryos subjected to zona thinning were compared with those of the hatching-defective embryos implanted without zona thinning. SETTING: University-based basic research laboratory. RESULTS: When hatching-defective embryos, created by destruction of 1/4 of the blastomeres at the four-cell stage, were implanted into pseudopregnant female mice, no pregnancies were established, and no implantation sites could be identified after transfer of 41 embryos. However, when hatching-defective embryos were subjected to zona thinning before ET, a normal rate of implantation was observed (18 implantation events of 49 embryos transferred). Fetuses appeared morphologically normal. The difference in implantation rates between hatching-defective embryos subjected to zona thinning and those not treated by zona thinning was highly significant by chi 2 analysis. CONCLUSION: Failure of embryos with cell numbers reduced by micromanipulation to implant and restoration of normal implantation and development after circumferential zona thinning demonstrates that the developmental defect of the manipulated embryos is confined to hatching, and thus, that this procedure establishes a useful model for defective hatching. High rates of implantation after circumferential thinning of the zona shows that this procedure of assisted hatching is highly effective. These findings have important implications for use of assisted hatching in human IVF because the assisted hatching procedure is safe and effective and increases the efficiency of the normal hatching mechanism. Thus, the procedure may be useful in a wide variety of circumstances for which assisted hatching by zona opening has been found to be ineffective.

Animals↗
PubMed1992

Bioeconomic evaluation of embryo transfer in beef production systems: I. Description of a biological model for steer production.

Concepts used to derive a deterministic model for evaluating embryo transfer for commercial steer production taking into consideration genetic merit for growth and mature size, herd feed supply, and recipient maternal environment are discussed. Genetic potential of an embryo is used to derive optimal growth rates that can be sustained by available herd feed per animal per day. Equations are provided for various measures of performance as functions of the feed, genotype of the embryo, and recipient maternal contribution. To assess the value of a particular line of embryos, interactions between genotype and nutrient environment are quantified, so that the benefits of embryos of high genetic merit are evaluated objectively. Product quality and weight are predicted from the model to provide a framework that will allow commercial beef producers to determine marketing strategies likely to result in optimal return.

Animal Feed↗
PubMed2003

Increased TNF-alpha expression in cultured mouse embryos exposed to teratogenic concentrations of glucose.

Diabetes-induced early embryonic death is accompanied by an increased expression of tumour necrosis factor alpha (TNF-alpha) in the embryonic microenvironment. The aim of the present study was to evaluate whether diabetes-induced embryopathic stress may also alter the expression of TNF-alpha produced by the embryo itself. As a model, whole postimplantation embryos were cultured for 24 h in a medium with high concentrations of glucose, one of the main diabetes-associated teratogenic metabolites. An anomaly such as an open neural tube was used as an end-point characterizing the glucose-induced teratogenic effect and the number of somites was counted to evaluate growth retardation induced by glucose. The expression of TNF-alpha (by immunohistochemistry), apoptosis (by TdT-mediated dUTP nick-end labelling; TUNEL) and the activity of caspases 3 and 8 (by a fluorometric assay) were evaluated in normal and malformed embryos. Ninety-seven per cent of the embryos exposed to 1300 mg glucose dl(-1) exhibited an open neural tube. The percentage of malformed embryos was smaller in media containing 800 and 500 mg glucose dl(-1) (68 and 37%, respectively) but it still exceeded significantly the value registered in embryos developing in a normoglycaemic medium (12%). In addition, a significant decrease in the number of somites was observed in embryos developing in media containing 1300 and 800 mg glucose dl(-1). Malformed embryos exhibited a greater number of nuclei that were positive in the TUNEL assay as well as a higher amount of active caspase 8 compared with normal embryos (with closed neural folds). TNF-alpha expression was detected in the neuroepithelial layer of the neural tube of the malformed embryos, whereas the expression of this cytokine was weak, if detectable, in normal embryos. Together, these findings indicate that TNF-alpha produced by the embryo may be involved in regulating the response of embryos to diabetes-generated embryopathic stress.

Abnormalities, Drug-Induced↗
PubMed2000

[Effects of endocrine disruptors on preimplantation embryo development].

Endocrine disruptors such as dioxins and bisphenol A(BPA) were detected in the human follicular fluids indicating that the reproductive organs have been exposed to these substances. To study possible effects of these chemicals on the germ cells and early embryos, we employed mouse preimplantation embryo development models. Dioxin(TCDD) and BPA both have positive and negative effects on embryo development in mice. The developmental rates were significantly decreased by exposure to concentrations of TCDD between 1 and 5 pM as compared with the control group. On the other hand, stimulatory effects of BPA were observed in the blastocyst formation rate at concentrations of 1 nM and 3 nM. The present study demonstrated that low concentration of both TCDD and BPA, not dose dependent manner, exert stimulatory or inhibitory effects on preimplantation embryo development. These alterations in low doses may be mediated by AhR or ER alpha and ER beta, present in mouse preimplantation embryos.

Animals↗