Complete replacement of the mitochondrial genotype in a Bos indicus calf reconstructed by nuclear transfer to a bos taurus oocyte

Due to the exclusively maternal inheritance of mitochondria, mitochondrial genotypes can be coupled to a particular nuclear genotype by continuous mating of founder females and their female offspring to males of the desired nuclear genotype. However, backcrossing is a gradual procedure that, apart from being lengthy, cannot ascertain that genetic and epigenetic changes will modify the original nuclear genotype. Animal cloning by nuclear transfer using host ooplasm carrying polymorphic mitochondrial genomes allows, among other biotechnology applications, the coupling of nuclear and mitochondrial genotypes of diverse origin within a single generation. Previous attempts to use Bos taurus oocytes as hosts to transfer nuclei from unrelated species led to the development to the blastocyst stage but none supported gestation to term. Our aim in this study was to determine whether B. taurus oocytes support development of nuclei from the closely related B. indicus cattle and to examine the fate of their mitochondrial genotypes throughout development. We show that indicus:taurus reconstructed oocytes develop to the blastocyst stage and produce live offspring after transfer to surrogate cows. We also demonstrate that, in reconstructed embryos, donor cell-derived mitochondria undergo a stringent genetic drift during early development leading, in most cases, to a reduction or complete elimination of B. indicus mtDNA. These results demonstrate that cross-subspecies animal cloning is a viable approach both for matching diverse nuclear and cytoplasmic genes to create novel breeds of cattle and for rescuing closely related endangered cattle.

Armazenamento, tolerância à dessecação e o potencial fisiológico de sementes de híbridos de milho e seus recíprocos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Variances of direct genetic effects, maternal genetic effects, and cytoplasmic inheritance effects for milk yield, fat yield, and fat percentage

Milk yield, fat yield, and fat percentage during the first three lactations were studied using New York Holsteins that were milked twice daily over a 305-d, mature equivalent lactation. Those data were used to estimate variances from direct and maternal genetic effects, cytoplasmic effects, sire by herd interaction, and cow permanent environmental effects. Cytoplasmic line was traced to the last female ancestor using DHI records from 1950 through 1991. Records were 138,869 lactations of 68,063 cows calving from 1980 through 1991. Ten random samples were based on herd code. Samples averaged 4926 dams and 2026 cytoplasmic lines. Model also included herd-year-seasons as fixed effects and genetic covariance for direct-maternal effects. Mean estimates of the effects of maternal genetic variances and direct-maternal covariances, as fractions of phenotypic variances, were 0.008 and 0.007 for milk yield, 0.010 and 0.010 for fat yield, and 0.006 and 0.025 for fat percentage, respectively. Average fractions of variance from cytoplasmic line were 0.011, 0.008, and 0.009 for milk yield, fat yield, and fat percentage. Removal of maternal genetic effects and covariance for maternal direct effects from the model increased the fraction of direct genetic variance by 0.014, 0.021, and 0.046 for milk yield, fat yield, and fat percentage; little change in the fraction was due to cytoplasmic line. Exclusion of cytoplasmic effects from the model increased the ratio of additive direct genetic variance to phenotypic variance by less than 2%. Similarly, when sire by herd interaction was excluded, the ratio of direct genetic variance to phenotypic variance increased 1% or less.

Hypercapnic ventilatory response of anesthetized female rats subjected to neonatal maternal separation: Insight into the origins of panic attacks?

Neonatal maternal separation (NMS) is a form of stress that interferes with the regulation of the stress response, an effect that predisposes to the emergence of panic and anxiety related disorders. We previously showed that at adulthood, awake female (but not male) rats subjected to NMS show a hypercapnic ventilatory response (HCVR; 5% CO(2)) that is 63% greater than controls (Genest et al., 2007). To understand the mechanisms underlying the sex-specific effects of NMS on the ventilatory response to CO(2), we used two different anesthetized female rat preparations to assess central CO(2) chemosensitivity and contribution of sensory afferents (stretch receptors and peripheral chemoreceptors) that influence the HCVR. Data show that anesthesia eliminated the respiratory phenotype observed previously in awake females and CO(2) chemosensitivity did not differ between groups. Finally, the assessment of the ovarian hormone levels across the oestrus cycle failed to reveal significant differences between groups. Since anesthesia did not affect the manifestation of NMS-related respiratory dysfunction in males (including the hypercapnic ventilatory response) (Kinkead et al., 2005; Dumont and Kinkead, 2010), we propose that the panic or anxiety induced by CO(2) during wakefulness is responsible for enhancement of the HCVR in NMS females. (C) 2011 Elsevier B.V. All rights reserved.

Efeito da linhagem citoplasmática sobre a produção de leite em bovinos da raça Caracu

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of data structure on the estimation of the additive genetic direct and maternal covariance for early growth traits in Nellore cattle

The objective of the present study was to investigate the effect of data structure on estimated genetic parameters and predicted breeding values of direct and maternal genetic effects for weaning weight (WW) and weight gain from birth to weaning (BWG), including or not the genetic covariance between direct and maternal effects. Records of 97,490 Nellore animals born between 1993 and 2006, from the Jacarezinho cattle raising farm, were used. Two different data sets were analyzed: DI_all, which included all available progenies of dams without their own performance; DII_all, which included DI_all + 20% of recorded progenies with maternal phenotypes. Two subsets were obtained from each data set (DI_all and DII_all): DI_1 and DII_1, which included only dams with three or fewer progenies; DI_5 and DII_5, which included only dams with five or more progenies. (Co)variance components and heritabilities were estimated by Bayesian inference through Gibbs sampling using univariate animal models. In general, for the population and traits studied, the proportion of dams with known phenotypic information and the number of progenies per dam influenced direct and maternal heritabilities, as well as the contribution of maternal permanent environmental variance to phenotypic variance. Only small differences were observed in the genetic and environmental parameters when the genetic covariance between direct and maternal effects was set to zero in the data sets studied. Thus, the inclusion or not of the genetic covariance between direct and maternal effects had little effect on the ranking of animals according to their breeding values for WW and BWG. Accurate estimation of genetic correlations between direct and maternal genetic effects depends on the data structure. Thus, this covariance should be set to zero in Nellore data sets in which the proportion of dams with phenotypic information is low, the number of progenies per dam is small, and pedigree relationships are poorly known. (c) 2012 Elsevier B.V. All rights reserved.

Efeito da linhagem citoplasmática sobre o peso ao nascer e o ganho médio diário na pré-desmama em bovinos da raça Caracu

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

INHERITANCE of CUCUMBER TOLERANCE TO ZUCCHINI YELLOW MOSAIC VIRUS

Studies were carried out in Brazil to study the inheritance of tolerance to Zucchini yellow mosaic virus (ZYMV) in cucumber cv. Formosa. This cultivar was individually crossed with two cucumber lines from different varietal types (L(b) from a Brazilian type, and L(j) from a Japanese type), both susceptible to the virus. Two experiments, one for each line, were separately carried out, where 6 treatments (parents, generations F1, F2 and F1BC1 for both parents) were evaluated in a randomized block design with 5 repetitions. Cotyledons of 2-week-old cucumber seedlings were inoculated with ZYMV. Only the plants that did not show symptoms up to 63 days post inoculation were considered as tolerant. A chi-square (chi(2)) analysis for assessing segregation from F2 and both F1BC1, led to the conclusion that the tolerance found in cv. Formosa is determined by a recessive gene.

Effect of cocaine on periadolescent rats with or without early maternal separation

Cocaine-induced behavioral sensitization and weight loss were investigated in periadolescent Wistar rats kept with their mothers or subjected to repeated maternal separation. Litters allocated to the separation procedure were placed in a temperature-controlled (33ºC) chamber for 3 h per day from postnatal day 6 (P6) to P20. Non-handled rats were left undisturbed until weaning. Treatments were started on P30-31 and the test was performed on P36-37. Animals received injections of saline or cocaine (10 mg/kg, sc) twice daily for 5 days. on day 6 all animals received saline. on day 7 animals were challenged with 10 mg/kg cocaine and their locomotion was evaluated in activity cages. A third group received saline throughout the 7-day period. Body weights were recorded on P30-31 and P36-37. Two-way ANOVA on body weights showed a main effect of treatment group (F(1,35) = 10.446, P = 0.003; N = 10-12). Non-handled rats treated with cocaine for 5 days gained significantly less weight, while no significant effect was observed in maternally separated rats. Two-way ANOVA revealed a main effect of drug treatment on locomotor activity (F(2,32) = 15.209, P<0.001; N = 6-8), but not on rearing condition (F(1,32)<0.001, P = 0.998). Animals pretreated with cocaine showed a clear behavioral sensitization relative to the saline group. No difference in the magnitude of sensitization was found between separated and non-handled animals. Only the effect of cocaine on weight gain was significantly affected by repeated episodes of early maternal separation during the pre-weaning period.

Maternal separation stress in male mice: long-term increases in alcohol intake

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of hypertension on maternal adaptations to pregnancy: experimental study on spontaneously hypertensive rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of Bauhinia forficata extract in diabetic pregnant rats: maternal repercussions

Bauhinia forficata, commonly known as paw-of-cow, is widely used in Brazil folk medicine for the treatment of Diabetes mellitus. The purposes of present study were to determine the repercussions of diabetes on the defense system against oxidative stress in pregnant female rats and to characterize the influence of the treatment with Bauhinia forficata extract on the antioxidant system, glycemic control, hepatic glycogen, cholesterol, triglycerides, total proteins and lipids. Virgin female Wistar rats were injected with 40 mg/kg streptozotocin (STZ) before mating. Oral administration of an aqueous extract of Bauhinia forficata leaves was given to non-diabetic and diabetic pregnant rats in 3 doses: 500 mg/kg from 0 to 4(th) day of pregnancy, 600 mg/kg from 5(th) to 14(th) day and 1000 mg/kg from 15(th) to 20(th) day. All the females were killed on the day 21 of pregnancy. A maternal blood sample was collected by venous puncture and the maternal liver was removed for biochemical measurement. The diabetic pregnant rats presented hyperglycemia, hyperlipemia, bypertriglyceridemia, hypercholesterolemia, hyperuricemia, decreased determinations of reduced glutathione (GSH) and superoxide dismutase (SOD). Treatment with B. forficata extract did not interfere in the albumin, total protein and lipid, triglyceride, cholesterol and SOD determinations. Increased hepatic glycogen, decreased uric acid concentration and increased GSH activity was observed. This last fact suggests that the plant may have some action on antioxidant defense system. However, the demonstration of the active component present in B. forficata responsible for its antioxidant effect and the increase in hepatic glycogen deserve further investigation.