Identification of optimal assisted aspiration conditions of oocytes for use in porcine in vitro maturation: a re-evaluation of the relationship between the cumulus oocyte complex and oocyte quality

The quality of porcine oocytes for use in IVF is commonly graded according to the number of layers of cumulus cells surrounding the oocyte; together these form the cumulus oocyte complex (COC). At least three compact layers of cumulus cells is regarded as important for efficient IVP. To test this, oocytes were scored according to cumulus investment, with grade A representing COCs with three or more cumulus layers including granulosa cell-cumulus oocyte complexes, grade B those with an intact corona radiata surrounded by another layer of cumulus cells and grades C and D representing COCs with lower cumulus cell investment. These oocytes were then monitored for in vitro maturation (IVM), as assessed by tubulin immunostaining for meiotic progression, the development of a cortical granule ring, and by glutathione levels. Results indicate that grading correlates closely with nuclear maturation and cytoplasmic maturation, suggesting that grading oocytes by cumulus investment is a reliable method to predict IVM success. Importantly, Grade A and B oocytes showed no significant differences in any measure and hence using a cut-off of two or more cumulus cell layers may be optimal. We also determined the effect of assisted aspiration for oocyte retrieval, comparing the effect of needle size and applied pressure on the retrieval rate. These data indicated that both variables affected oocyte recovery rates and the quality of recovered oocytes. In combination, these experiments indicate that grade A and B oocytes have a similar developmental potential and that the recovery of oocytes of these grades is maximised by use of an 18-gauge needle and 50mmHg aspiration pressure

Mapeamento de locos de características quantitativas no cromossomo 6, associados às características de carcaça e de órgãos internos de suínos Mapping of quantitative trait loci for carcass traits and internal organs in swine chromosome 6

Com o objetivo de mapear locos de características quantitativas (QTLs) associados às características de carcaça e de órgão internos, uma população composta de 550 animais F2 foi produzida a partir do intercruzamento da geração F1, obtida pelo cruzamento divergente de dois machos da raça nativa brasileira Piau com 18 fêmeas comerciais. Os animais foram genotipados para 13 marcadores microssatélites distribuídos no cromossomo 6. As características avaliadas foram: comprimento de carcaça pelos métodos brasileiro e americano, peso e rendimento de carcaça, espessura de toucinho na região da copa, espessura de toucinho imediatamente após a última costela, espessura de toucinho entre a última e a penúltima vértebra lombar, menor espessura de toucinho na região acima da última vértebra lombar, espessura de toucinho imediatamente após a última costela, a 6,5 cm da linha dorso-lombar, espessura de toucinho média (geral = estimada a partir da média de todas as espessuras de toucinho citadas anteriormente; e dorso-lombar = estimada a partir das espessuras de toucinho tomadas na linha dorso-lombar do animal), espessura de bacon, profundidade de lombo, área de olho-de-lombo, pesos de órgãos internos (coração, pulmões, fígado, baço e rim) e comprimento de intestino. Utilizou-se o método de regressão por intervalo de mapeamento por meio do programa QTL Express. Foram encontrados QTLs sugestivos para as características de comprimento de carcaça pelo método brasileiro e espessura de bacon e QTL significativo para peso do rim. Nos intervalos dos picos da estatística F em que se encontraram QTLs sugestivos, devem ser incluídos mais marcadores para se confirmar a real presença de QTL.<br>A total of the 550 F2 animals produced by divergent cross using two sires of the native Brazilian breed named Piau and 18 commercial dams were genotyped for 13 microsatellite markers in swine chromosome 6. The traits evaluated were: carcass length by the Brazilian carcass classification method, carcass length by the American carcass classification method, carcass weight, carcass yield, higher backfat thickness at last 2nd-3rd thoracic vertebrae, backfat thickness after last rib, backfat thickness between last 1st-2nd lumbar vertebrae, lower backfat thickness after last lumbar vertebrae, backfat thickness after last rib, at 6,5 cm from the midline, average backfat thickness (global = estimated by average of all backfat thickness; and midline = estimated by average of midline backfat thickness), bacon depth, loin depth and loin eye area; internal organs weight (heart, lungs, liver, kidney and spleen), and intestine length. Data were analyzed by multiple regression using the interval mapping method from the QTL Express software. Suggestive QTL were found for carcass length by the Brazilian method and bacon depth and a significant one for kidney weight. Additional markers should be included in the peak intervals of the suggestive F-values to confirm the presence of QTL

Regression-based quantitative trait loci mapping: robust, efficient and effective

Regression has always been an important tool for quantitative geneticists. The use of maximum likelihood (ML) has been advocated for the detection of quantitative trait loci (QTL) through linkage with molecular markers, and this approach can be very effective. However, linear regression models have also been proposed which perform similarly to ML, while retaining the many beneficial features of regression and, hence, can be more tractable and versatile than ML in some circumstances. Here, the use of linear regression to detect QTL in structured outbred populations is reviewed and its perceived shortfalls are revisited. It is argued that the approach is valuable now and will remain so in the future