Unraveling trafficking of the kidney anion exchanger 1 in polarized MDCK epithelial cells.

Kidney anion exchanger 1 (kAE1) is a membrane glycoprotein expressed at the basolateral membrane of type A intercalated cells in the kidney collecting tubule. Mutations occurring in the gene encoding this protein can give rise to distal renal tubular acidosis (dRTA), a disease characterized by an impaired urine acidification, nephrocalcinosis, and renal failure. Here we review how the study of dRTA mutants in polarized epithelial cells has shed light on the cellular mechanisms resulting in this renal disease.     

On the streets of San Francisco: highlights from the ISSCR Annual Meeting 2010

The 2010 Annual Meeting of the International Society for Stem Cell Research (ISSCR) was held in San Francisco in June with an exciting program covering a wealth of stem cell research from basic science to clinical research.     

The source of laterally transferred genes in bacterial genomes

Background  

Laterally transferred genes have often been identified on the basis of compositional features that distinguish them from ancestral genes in the genome. These genes are usually A+T-rich, arguing either that there is a bias towards acquiring genes from donor organisms having low G+C contents or that genes acquired from organisms of similar genomic base compositions go undetected in these analyses.     

Interacting effects of age,density, and weather on survival and current reproduction for a large mammal

Individual‐based study of natural populations allows for accurate and precise estimation of fitness components and the extent to which they might vary with ecological conditions. By tracking the fates of all 701 horses known to have lived on Sable Island, Canada, from 2009 to 2013 (where there is no predation, human interference, or interspecific competition for food), we present a detailed analysis of structured population dynamics with focus on interacting effects of intraspecific competition and weather on reproduction and survival. Annual survival of adult females (0.866 ± 0.107 [ ± SE]) was lower than that of 3‐year‐olds (0.955 ± 0.051), although annual fecundity (producing a foal in a year that was observed during our census) was higher in adults (0.616 ± 0.023) compared to 3‐year‐olds (0.402 ± 0.054). Milder winters and lower densities during gestation increased fecundity. Density negatively impacted survival for all age and sex categories; however, highest adult female survival was observed during high‐density years coupled with a harsh winter, the result expected if pregnancy loss during winter or loss of foals in spring improved survival. Three‐year‐old females, which reproduced at lower rates, experienced higher survival than adults. Our results contrast with a previous study of feral horses that suggested recently feral ungulates might be artificially selected to reproduce even when costs to survival are high. In part, this may be because of the comparably long history of feralization (250 years; at least 25 generations) for Sable Island horses.     

Dynamic management of genetic resources: maintenance of outcrossing in experimental metapopulations of a predominantly inbreeding species

Dynamic management of genetic resources inpredominantly inbreeding species requiresincreased levels of outcrossing to limit theloss of genetic variation due to smallereffective sizes and to favour the emergence ofnew genetic combinations. Here, we show thatoutcrossing rates can artificially bepermanently increased in experimental evolvingplant metapopulations, using Arabidopsisthaliana as a model. We introducedmale-sterility genes and used an adequatemanagement of the resulting female plants tomodify the outcrossing rates. As expected, theincrease in outcrossing resulted in lowerlevels of heterozygote deficiency (F _is) than observed in natural populationsof A. thaliana and therefore in themaintenance of potentially higher levels ofgenetic variation. An additional selectiveadvantage for females' offspring, due to theproduction of larger seeds by females and apossible heterosis effect, furthermore led tosmaller F _is than expected from therealized outcrossing rate. This selectiveadvantage also resulted in an increase infemale frequency, especially in metapopulationswith large population sizes, creating anon-causal negative correlation between femalefrequency and heterozygote deficiency.     

Vanadium-dependent iodoperoxidases in Laminaria digitata, a novel biochemical function diverging from brown algal bromoperoxidases

The brown alga Laminaria digitata features a distinct vanadium-dependent iodoperoxidase (vIPO) activity, which has been purified to electrophoretic homogeneity. Steady-state analyses at pH 6.2 are reported for vIPO (K _m ^I– =2.5 mM; k _cat ^I– =462 s^–1) and for the previously characterised vanadium-dependent bromoperoxidase in L. digitata (K _m ^I– =18.1 mM; k _cat ^I– =38 s^–1). Although the vIPO enzyme specifically oxidises iodide, competition experiments with halides indicate that bromide is a competitive inhibitor with respect to the fixation of iodide. A full-length complementary ANA (cDNA) was cloned and shown to be actively transcribed in L. digitata and to encode the vIPO enzyme. Mass spectrometry analyses of tryptic digests of vIPO indicated the presence of at least two very similar proteins, in agreement with Southern analyses showing that vIPOs are encoded by a multigenic family in L. digitata. Phylogenetic analyses indicated that vIPO shares a close common ancestor with brown algal vanadium-dependent bromoperoxidases. Based on a three-dimensional structure model of the vIPO active site and on comparisons with those of other vanadium-dependent haloperoxidases, we propose a hypothesis to explain the evolution of strict specificity for iodide in L. digitata vIPO.The nucleotide sequence reported in this paper has been submitted to the EBI Data Bank with accession no. AJ619804.     

Alterations of adenylyl cyclase and G proteins in aortocaval shunt-induced heart failure

Unlike most other experimental models of congestive heart failure, the volume overload model induced by aortocaval shunt (AVS) in rats was found to exhibit enhanced beta-adrenoceptor (beta-AR) signaling. To study whether the adenylyl cyclase (AC)-G protein system is involved in such a change, we examined cardiac AC activity and protein content as well as G(s)alpha and G(i)alpha activities, protein contents, and mRNA levels in both left (LV) and right (RV) ventricles at the failing stage (16 wk after surgery). Basal and forskolin-stimulated AC activities were significantly increased in both LV and RV from the failing hearts; this change was associated with an upregulation of type V/VI AC protein. In contrast to 5'-guanylyl imidodiphosphate and NaF, the stimulatory effect of isoproterenol on AC was increased in the failing heart. Although G(s)alpha and G(i)alpha protein contents in the failing hearts were not altered, the mRNA level for G(s)alpha was decreased by 20% and that for G(i)alpha was increased by 20%. In addition, the activity of G(s)alpha, but not G(i)alpha, as assessed by toxin-catalyzed ADP ribosylation, was significantly decreased in the failing heart. Losartan and imidapril treatments improved cardiac function and attenuated alterations in mRNA levels for G(s)alpha and G(i)alpha proteins, as well as G(s)alpha activity, without affecting changes in AC protein content or activities in heart failure due to volume overload. These data suggest that increased AC activity may contribute to the enhanced beta-AR signaling in the AVS model of heart failure, whereas alterations in gene expression for G proteins may be of an adaptive nature at this stage of heart failure.     

Proteomic analysis of nuclear proteins from proliferative and differentiated human colonic intestinal epithelial cells

Self-renewing tissues such as the intestine contain progenitor proliferating cells which subsequently differentiate. Cell proliferation and differentiation involve gene regulation processes which take place in the nucleus. A human intestinal epithelial cell line model (Caco2/TC7) which reproduces these dynamic processes has been used to perform proteomic studies on nuclear proteins. Nuclei from Caco2/TC7 cells at proliferative and differentiated stages were purified by subcellular fractionation. After two-dimensional gel electrophoresis separation and ruthenium staining, 400 protein spots were detected by image analysis. Eighty-five spots corresponding to 60 different proteins were identified by matrix-assisted laser desorption/ionization mass spectrometry in nuclei from proliferative cells. Comparison of nuclear proteomes from proliferative or differentiated cells by differential display resulted in the identification of differentially expressed proteins such as nucleolin, hnRNP A2/B1 and hnRNP A1. By using Western blot analysis, we found that the expression and number of specific isoforms of these nuclear proteins decreased in differentiated cells. Immunocytochemistry experiments also showed that in proliferative cells nucleolin was distributed in nucleoli-like bodies. In contrast, hnRNPs A2/B1 and A1 were dispersed throughout the nucleus. This study of the nuclear proteome from intestinal epithelial cells represents the first step towards the establishment of a protein database which will be a valuable resource in future studies on the differential expression of nuclear proteins in response to physiological, pharmacological and pathological modulations.