Expression of DLK1 and MEG3 genes in porcine tissues during postnatal development

The Drosophila-like homolog 1 (DLK1), a transmembrane signal protein similar to other members of the Notch/Delta/Serrate family, regulates the differentiation process in many types of mammalian cells. Callipyge sheep and DLK1 knockout mice are excellent examples of a fundamental role of the gene encoding DLK1 in muscle growth and fat deposition. DLK1 is located within co-regulated imprinted clusters (the DLK1/DIO3 domain), along with other imprinted genes. Some of these, e.g. the RNA coding MEG3 gene, presumedly interfere with DLK1 transcription. The aim of our study was to analyze DLK1 and MEG3 gene expression in porcine tissues (muscle, liver, kidney, heart, brain stem) during postnatal development. The highest expression of both DLK1 and MEG3 variant 1 (MEG3 var.1) was observed in the brain-stem and muscles, whereas that of MEG3 variant 2 (MEG3var.2) was the most abundant in muscles and the heart. During development (between 60 and 210 days of age) expression of analyzed genes was down-regulated in all the tissues. An exception was the brain- stem, where there was no significant change in MEG3 (both variants) mRNA level, and relatively little decline (2-fold) in that of DLK1 transcription. This may indicate a distinct function of the DLK1 gene in the brain-stem, when compared with other tissues

Tolerância de rizóbios de diferentes procedências ao zinco, cobre e cádmio Tolerance of rhizobia genera from different origins to zinc, copper and cadmium

Sessenta estirpes/isolados dos gêneros Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium e Azorhizobium, procedentes de diferentes locais (Mata Atlântica, Amazônia, culturas agrícolas e experimentos com metais pesados) e de espécies hospedeiras pertencentes às subfamílias Papilionoideae, Mimosoideae e Caesalpinoideae, foram avaliadas quanto à tolerância a Zn, Cu e Cd em meio YMA modificado pela adição de tampões biológicos (HEPES e MES) e suplementados com Cu (0 a 60 mg L-1), Cd (0 a 60 mg L-1) e Zn (0 a 1.000 mg L-1). Mediante padrões de crescimento atribuídos às culturas nas diferentes concentrações dos metais, avaliaram-se as concentrações máximas toleradas e as doses tóxicas destes metais para redução de crescimento em 25% (DT25) e 50% (DT50). Não houve influência da procedência na concentração máxima de metal tolerada. A ordem de sensibilidade aos metais, considerando-se as concentrações máximas toleradas, foi Azorhizobium > Rhizobium = Mesorhizobium = Sinorhizobium > Bradyrhizobium. A DT25 e a DT50 foram úteis para diferenciarem estirpes/isolados de um mesmo gênero, que atingiram a mesma concentração máxima tolerada a Zn, Cu e Cd. A ordem de toxicidade dos metais estudados foi Cu > Cd > Zn.<br>Sixty strains/isolates of the genera Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium and Azorhizobium, isolated from different hosts (legume subfamilies: Papilionoideae, Mimosoideae and Caesalpinoideae) and location (Atlantic Forest, Amazon region, crop plantings and heavy metal experiments), were evaluated for Zn, Cu and Cd tolerance in YMA medium modified by the addition of biological buffers (HEPES and MES) and supplemented with Cu (0 to 60 mg L-1), Cd (0 to 60 mg L-1), and Zn (0 to 1,000 mg L-1)sulphates. Growth standards were applied to evaluate rhizobia cultures growth at different metal concentrations, allowing evaluation of highest tolerated concentrations of Zn, Cu, and Cd and the toxic doses (concentrations) of these metals, which reduce the growth standard in 25% (DT25) and 50% (DT50). It was verified that there was no influence of the origin (host and location) on the highest tolerated concentration of each metal; the order of sensitivity to heavy metals, considering the highest tolerable concentrations, was Azorhizobium > Rhizobium = Mesorhizobium = Sinorhizobium > Bradyrhizobium. The DT25 and the DT50 were useful to differentiate strain/isolates of the same genus, which reached the same highest tolerated concentration to Zn, Cu, and Cu; and the order of toxicity was Cu > Cd > Zn