Skeletal muscle overexpression of sAnk1.5 in transgenic mice does not predispose to type 2 diabetes.

Genome-wide association studies (GWAS) and cis-expression quantitative trait locus (cis-eQTL) analyses indicated an association of the rs508419 single nucleotide polymorphism (SNP) with type 2 diabetes (T2D). rs508419 is localized in the muscle-specific internal promoter (P2) of the ANK1 gene, which drives the expression of the sAnk1.5 isoform. Functional studies showed that the rs508419 C/C variant results in increased transcriptional activity of the P2 promoter, leading to higher levels of sAnk1.5 mRNA and protein in skeletal muscle biopsies of individuals carrying the C/C genotype. To investigate whether sAnk1.5 overexpression in skeletal muscle might predispose to T2D development, we generated transgenic mice (TgsAnk1.5/+) in which the sAnk1.5 coding sequence was selectively overexpressed in skeletal muscle tissue. TgsAnk1.5/+ mice expressed up to 50% as much sAnk1.5 protein as wild-type (WT) muscles, mirroring the difference reported between individuals with the C/C or T/T genotype at rs508419. However, fasting glucose levels, glucose tolerance, insulin levels and insulin response in TgsAnk1.5/+ mice did not differ from those of age-matched WT mice monitored over a 12-month period. Even when fed a high-fat diet, TgsAnk1.5/+ mice only presented increased caloric intake, but glucose disposal, insulin tolerance and weight gain were comparable to those of WT mice fed a similar diet. Altogether, these data indicate that sAnk1.5 overexpression in skeletal muscle does not predispose mice to T2D susceptibility.

Biochemical and morphological comparison of microsomal preparations from rat, quail, trout, mussel, and water flea.

Differential centrifugation methods already in use were applied to purify rat, quail, and trout liver microsomes and modified as necessary to purify microsomes from mussel digestive gland and whole water flea. All these microsomal preparations were comparatively characterized with respect to protein and RNA content, levels of markers of subcellular contaminants, ultrastructural morphology, differential spectra of cytochromes P-450 and b5, monoxygenase activity, and in vitro metabolism of p-dichlorobenzene. Yields of microsomal proteins of the tested organisms differed widely, with mussel showing the lowest yield. Very low levels of nuclear and mitochondrial contaminants were detected in all microsomal preparations, but cell membrane contaminants were clearly present in most preparations. Daphnia microsomes were significantly contaminated by plasma membranes, and hepatopancreas microsomes contained significant amounts of partially disrupted secretory granules and plasma membrane. From a qualitative standpoint differential spectra of cytochrome b5 were very similar for all the preparations, whereas cytochrome P-450 spectra were largely dependent on the microsomal preparation as well as on the assay method used. The content of cytochrome P-450 was highest for rat liver microsomes and very low or absent in Daphnia and mussel preparations; the range of cytochrome b5 contents was much narrower. Significant differences were observed among monoxygenase activities of the different preparations. In Daphnia and mussel microsomes, aniline hydroxylase was absent and benzo[a]pyrene hydroxylase activity was much lower than in rat and quail microsomes. Benzo[a]pyrene hydroxylase activity of trout liver microsomes was similar to that of rat and quail microsomes, whereas hydroxylation of substrates which in rat liver are preferentially metabolized by phenobarbital-inducible forms of cytochrome P-450 was much lower in trout microsomes.