This study aimed to investigate the mode of action of cadmium (Cd) toxicity at cell level, especially at early stages of plant exposure. Tomato seedlings were cultivated in growth media containing from 0.1 to 70 µM CdCl2 for 24 h. Mitotic index, chromosome abnormality, DNA integrity and organization of tubulin-based structures were assessed in root cells. As higher the Cd concentration in the growth media, higher was the DNA damage intensity and the occurrence of chromosomal abnormalities that included chromosome lost, bridges, stickiness, C-metaphase and polyploidy. The profile of chromosomal aberrations also varied with elevated Cd concentration, being observed increases in the frequency of chromosome stickiness. The mitotic index was reduced at the lowest Cd concentration, but such reduction was statistically similar to that detected at the highest concentration, suggesting that mitotic depression is a rapid outcome and, at same time, a Cd-induced effect that is limited at the first 24 h of direct root exposure to this metal. Under exposure to 20 µM CdCl2, heterogenous distribution of the spindle fibers, formation of two spindle complexes in both of the cell poles, absence of centrosome center, polarization of the spindle fibers during cell division, and non-uniform tubulin deposition in microtubule and phragmoplast were noticed. The results indicate that the tubulin-dependent components of cytoskeleton are Cd targets, and the sensitivity of tubulin-based structures to Cd exposure depends on cell cycle phase. Moreover, DNA damage intensity and chromosomal abnormality profile can be employed as markers of Cd toxicity level.
Cadmium/toxicity , Cell Cycle/drug effects , Chromosomal Instability/drug effects , Cytoskeleton/drug effects , Soil Pollutants/toxicity , Solanum lycopersicum/drug effects , Seedlings/drug effects
Repulsive guidance molecules (RGMs) compose a family of glycosylphosphatidylinositol (GPI)-anchored axon guidance molecules and perform several functions during neural development. New evidence has suggested possible new roles for these axon guidance molecules during skeletal muscle development, which has not been investigated thus far. In the present study, we show that RGMa, RGMb and RGMc are all induced during skeletal muscle differentiation in vitro. Immunolocalization performed on adult skeletal muscle cells revealed that RGMa, RGMb and RGMc are sarcolemmal proteins. Additionally, RGMa was found to be a sarcoplasmic protein with a surprisingly striated pattern. RGMa colocalization with known sarcoplasmic proteins suggested that this axon guidance molecule is a skeletal muscle sarcoplasmic protein. Western blot analysis revealed two RGMa fragments of 60 and 33 kDa, respectively, in adult skeletal muscle samples. RGMa phenotypes in skeletal muscle cells (C2C12 and primary myoblasts) were also investigated. RGMa overexpression produced hypertrophic cells, whereas RGMa knockdown resulted in the opposite phenotype. RGMa knockdown also blocked myotube formation in both skeletal muscle cell types. Our results are the first to show an axon guidance molecule as a skeletal muscle sarcoplasmic protein and to include RGMa in a system that regulates skeletal muscle cell size and differentiation.
Muscle Fibers, Skeletal/metabolism , Muscle Proteins/metabolism , Muscle, Skeletal/metabolism , Animals , Cell Differentiation/physiology , Cell Enlargement , Hypertrophy/metabolism , Male , Membrane Proteins/metabolism , Mice , Muscle Development/physiology , Muscle, Skeletal/pathology , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neurogenesis/physiology
Two functional and positional candidate genes were selected in a region of chicken chromosome 1 (GGA1), based on their biological roles, and also where several quantitative trait loci (QTL) have been mapped and associated with performance, fatness and carcass traits in chickens. The insulin-like growth factor 1 (IGF1) gene has been associated with several physiological functions related to growth. The lysine (K)-specific demethylase 5A (KDM5A) gene participates in the epigenetic regulation of genes involved with the cell cycle. Our objective was to find associations of selected single-nucleotide polymorphisms (SNPs) in these genes with performance, fatness and carcass traits in 165 F(2) chickens from a resource population. In the IGF1 gene, 17 SNPs were detected, and in the KDM5A gene, nine SNPs were detected. IGF1 SNP c.47673G > A was associated with body weight and haematocrit percentage, and also with feed intake and percentages of abdominal fat and gizzard genotype × sex interactions. KDM5A SNP c.34208C > T genotype × sex interaction affected body weight, feed intake, percentages of abdominal fat (p = 0.0001), carcass, gizzard and haematocrit. A strong association of the diplotype × sex interaction (p < 0.0001) with abdominal fat was observed, and also associations with body weight, feed intake, percentages of carcass, drums and thighs, gizzard and haematocrit. Our findings suggest that the KDM5A gene might play an important role in the abdominal fat deposition in chickens. The IGF1 and KDM5A genes are strong candidates to explain the QTL mapped in this region of GGA1.
Body Weight/genetics , Chickens/anatomy & histology , Chickens/genetics , Insulin-Like Growth Factor I/genetics , Retinoblastoma-Binding Protein 2/genetics , Adipose Tissue/metabolism , Animals , Base Sequence , Biometry , Female , Genotype , Hematocrit , Male , Polymorphism, Single Nucleotide , Quantitative Trait Loci , Sequence Analysis, DNA
Quantitative trait loci (QTL) mapping in livestock allows the identification of genes that determine the genetic variation affecting traits of economic interest. We analyzed the birth weight and weight at 60 days QTL segregating on bovine chromosome BTA14 in a F2 resource population using genotypes produced from seven microsatellite markers. Phenotypes were derived from 346 F2 progeny produced from crossing Bos indicus Gyr x Holstein Bos taurus F1 parents. Interval analysis to detect QTL for birth weight revealed the presence of a QTL (p < 0.05) at 1 centimorgan (cM) from the centromere with an additive effect of 1.210 ± 0.438 kg. Interval analysis for weight at 60 days revealed the presence of a QTL (p < 0.05) at 0 cM from the centromere with an additive effect of 2.122 ± 0.735 kg. The region to which the QTL were assigned is described in the literature as responsible for some growth traits, milk yield, milk composition, fat deposition and has also been related to reproductive traits such as daughter pregnancy rate and ovulation rate. The effects of the QTL described on other traits were not investigated.
Os programas de melhoramento genético de frangos de corte que buscam máxima velocidade de ganho de peso, alta eficiência alimentar, alta viabilidade, maior rendimento de carcaça e menor deposição de gordura podem desencadear algumas síndromes fisiológicas, dentre as quais destacam-se o estresse calórico, a morte súbita e a ascite. A ascite se enquadra no conceito das síndromes multifatoriais, uma vez que sua manifestação ocorre quando certos fatores genéticos e ambientais atuam em conjunto determinando o processo. As limitações anatômica e fisiológica da circulação sanguínea nos pulmões provocam a síndrome de hipertensão pulmonar (PHS); esta pode provocar grande acúmulo de fluido na cavidade abdominal, quadro este denominado de ascite. Ocorre redução da eficiência da circulação sangüínea, levando as aves à morte por hipóxia, predominantemente no período entre 30 e 40 dias de idade. Uma vez desencadeado o processo ascítico, a ave dificilmente é aproveitada no abate já que a mesma restringe o consumo de alimento, ganhando menos peso. Adicionalmente, a carcaça apresenta aumento do volume da cavidade abdominal e conseqüente congestão dos órgãos internos. A descrição da fisiologia, medidas de avaliação e perspectivas são apresentadas neste trabalho.
