Ancestral bias in the Hras1 gene and distal Chromosome 7 among inbred mice

Inbred strains of mice vary in their frequency of liver tumors initiated by a mutation in the Hras1 (H-ras) proto-oncogene. We sequenced 4.5 kb of the Hras1 gene on distal Chr 7 in a diverse set of 12 commonly used laboratory inbred strains of mice and detected no sequence variation to account for strain-specific differences in Hras1 mutation prevalence. Furthermore, the Hras1 sequence is essentially monoallelic for an ancestral gene derived from the M. m. domesticus species. To determine if the monoallelism and associated low rate of polymorphism are unique to Hras1 or representative of the general chromosomal locale, we extended the sequence analysis to 12 genes in the final 8 Mb of distal Chr 7. A region of at least 2.5 Mb that encompasses several genes, including Hras1 and the H19/Igf2 loci, demonstrates virtually no sequence variation. The 12 inbred strains share one dominant haplotype derived from the M. m. domesticus allele. Chromosomal regions flanking the monoallelic segment exhibit a significantly higher rate of variation and multiple haplotypes, a majority of which are attributed to M. m. domesticus or M. m. musculus ancestry

The PPCD1 Mouse: Characterization of a Mouse Model for Posterior Polymorphous Corneal Dystrophy and Identification of a Candidate Gene

The PPCD1 mouse, a spontaneous mutant that arose in our mouse colony, is characterized by an enlarged anterior chamber resulting from metaplasia of the corneal endothelium and blockage of the iridocorneal angle by epithelialized corneal endothelial cells. The presence of stratified multilayered corneal endothelial cells with abnormal patterns of cytokeratin expression are remarkably similar to those observed in human posterior polymorphous corneal dystrophy (PPCD) and the sporadic condition, iridocorneal endothelial syndrome. Affected eyes exhibit epithelialized corneal endothelial cells, with inappropriate cytokeratin expression and proliferation over the iridocorneal angle and posterior cornea. We have termed this the “mouse PPCD1” phenotype and mapped the mouse locus for this phenotype, designated “Ppcd1”, to a 6.1 Mbp interval on Chromosome 2, which is syntenic to the human Chromosome 20 PPCD1 interval. Inheritance of the mouse PPCD1 phenotype is autosomal dominant, with complete penetrance on the sensitive DBA/2J background and decreased penetrance on the C57BL/6J background. Comparative genome hybridization has identified a hemizygous 78 Kbp duplication in the mapped interval. The endpoints of the duplication are located in positions that disrupt the genes Csrp2bp and 6330439K17Rik and lead to duplication of the pseudogene LOC100043552. Quantitative reverse transcriptase-PCR indicates that expression levels of Csrp2bp and 6330439K17Rik are decreased in eyes of PPCD1 mice. Based on the observations of decreased gene expression levels, association with ZEB1-related pathways, and the report of corneal opacities in Csrp2bptm1a(KOMP)Wtsi heterozygotes and embryonic lethality in nulls, we postulate that duplication of the 78 Kbp segment leading to haploinsufficiency of Csrp2bp is responsible for the mouse PPCD1 phenotype. Similarly, CSRP2BP haploinsufficiency may lead to human PPCD

Predominant modifier of extreme liver cancer susceptibility in C57BR/cdJ female mice localized to 6 Mb on chromosome 17

Sex hormones influence the susceptibility of inbred mice to liver cancer. C57BR/cdJ (BR) females are extremely susceptible to spontaneous and chemically induced liver tumors, in part due to a lack of protection against hepatocarcinogenesis normally offered by ovarian hormones. BR males are also moderately susceptible, and the susceptibility of both sexes of BR mice to liver tumors induced with N,N-diethylnitrosamine relative to the resistant C57BL/6J (B6) strain is caused by two loci designated Hcf1 and Hcf2 (hepatocarcinogenesis in females) located on chromosomes 17 and 1, respectively. The Hcf1 locus on chromosome 17 is the predominant modifier of liver cancer in BR mice. To validate the existence of this locus and investigate its potential interaction with Hcf2, congenic mice for each region were generated. Homozygosity for the B6.BR(D17Mit164-D17Mit2) region resulted in a 4-fold increase in liver tumor multiplicity in females and a 4.5-fold increase in males compared with B6 controls. A series of 16 recombinants covering the entire congenic region was developed to further narrow the area containing Hcf1. Susceptible heterozygous recombinants demonstrated a 3- to 7-fold effect in females and a 1.5- to 2-fold effect in males compared with B6 siblings. The effect in susceptible lines completely recapitulated the susceptibility of heterozygous full-length chromosome 17 congenics and furthermore narrowed the location of the Hcf1 locus to a single region of the chromosome from 30.05 to 35.83 Mb

A review of elliptical and disc galaxy structure, and modern scaling laws

A century ago, in 1911 and 1913, Plummer and then Reynolds introduced their models to describe the radial distribution of stars in `nebulae'. This article reviews the progress since then, providing both an historical perspective and a contemporary review of the stellar structure of bulges, discs and elliptical galaxies. The quantification of galaxy nuclei, such as central mass deficits and excess nuclear light, plus the structure of dark matter halos and cD galaxy envelopes, are discussed. Issues pertaining to spiral galaxies including dust, bulge-to-disc ratios, bulgeless galaxies, bars and the identification of pseudobulges are also reviewed. An array of modern scaling relations involving sizes, luminosities, surface brightnesses and stellar concentrations are presented, many of which are shown to be curved. These 'redshift zero' relations not only quantify the behavior and nature of galaxies in the Universe today, but are the modern benchmark for evolutionary studies of galaxies, whether based on observations, N-body-simulations or semi-analytical modelling. For example, it is shown that some of the recently discovered compact elliptical galaxies at 1.5 < z < 2.5 may be the bulges of modern disc galaxies.Comment: Condensed version (due to Contract) of an invited review article to appear in "Planets, Stars and Stellar Systems"(www.springer.com/astronomy/book/978-90-481-8818-5). 500+ references incl. many somewhat forgotten, pioneer papers. Original submission to Springer: 07-June-201

Mutations in Protein-Binding Hot-Spots on the Hub Protein Smad3 Differentially Affect Its Protein Interactions and Smad3-Regulated Gene Expression

Hub proteins are connected through binding interactions to many other proteins. Smad3, a mediator of signal transduction induced by transforming growth factor beta (TGF-β), serves as a hub protein for over 50 protein-protein interactions. Different cellular responses mediated by Smad3 are the product of cell-type and context dependent Smad3-nucleated protein complexes acting in concert. Our hypothesis is that perturbation of this spectrum of protein complexes by mutation of single protein-binding hot-spots on Smad3 will have distinct consequences on Smad3-mediated responses.We mutated 28 amino acids on the surface of the Smad3 MH2 domain and identified 22 Smad3 variants with reduced binding to subsets of 17 Smad3-binding proteins including Smad4, SARA, Ski, Smurf2 and SIP1. Mutations defective in binding to Smad4, e.g., D408H, or defective in nucleocytoplasmic shuttling, e.g., W406A, were compromised in modulating the expression levels of a Smad3-dependent reporter gene or six endogenous Smad3-responsive genes: Mmp9, IL11, Tnfaip6, Fermt1, Olfm2 and Wnt11. However, the Smad3 mutants Y226A, Y297A, W326A, K341A, and E267A had distinct differences on TGF-β signaling. For example, K341A and Y226A both reduced the Smad3-mediated activation of the reporter gene by ∼50% but K341A only reduced the TGF-β inducibilty of Olfm2 in contrast to Y226A which reduced the TGF-β inducibility of all six endogenous genes as severely as the W406A mutation. E267A had increased protein binding but reduced TGF-β inducibility because it caused higher basal levels of expression. Y297A had increased TGF-β inducibility because it caused lower Smad3-induced basal levels of gene expression.Mutations in protein binding hot-spots on Smad3 reduced the binding to different subsets of interacting proteins and caused a range of quantitative changes in the expression of genes induced by Smad3. This approach should be useful for unraveling which Smad3 protein complexes are critical for specific biological responses

Effects of changing from full range of motion to partial range of motion on squat kinetics

It is commonplace for people involved in recreational weight training to limit squat depth to lift heavier loads. This study compares differences in movement kinetics when squatting in the full range of motion (FROM) vs. partial range of motion (PROM). Ten men with a 1-year minimum of resistance training attended 4 sessions each comprising 4 sets of squats following one of FROM for 10 repetitions (FROM10) at an intensity of 67% 1 repetition maximum (1RM) FROM squat, PROM for 10 repetitions (PROM10) at 67% 1RM PROM squat, FROM for 5 repetitions (FROM5) at 83% FROM squat or PROM for 5 repetitions (PROM5) at 83% 1RM PROM squat. Movement velocity was not specified. Squat kinetics data were collected using an optical encoder. Differences between conditions were analyzed by repeated-measures analysis of variance and expressed as mean differences and standardized (Cohen) effect sizes with 95% confidence limits. The PROM5 power was substantially more than the PROM10 (98 W, -21 to 217; mean, lower and upper 95% confidence limits), FROM5 (168 W, 47-289), and FROM10 (255 W, 145-365). The force produced during PROM5 was substantially more than PROM10 (372 N, 254-490), FROM5 (854 N, 731-977), and FROM10 (1,069 N, 911-1227). The peak velocity produced during FROM10 was substantially more than FROM5 (0.105 m&middot;s(-1), 0.044-0.166), PROM10 (0.246 m&middot;s(-1), 0.167-0.325), and PROM5 (0.305 m&middot;s(-1), 0.228-0.382). The FROM5 was substantially more than FROM10 (86 J, 59-113), PROM5 (142 J, 90-194), and PROM10 (211 J, 165-257). Therefore, either range of motion can have practical implications in designing resistance training programs depending on if the training goal is related to power and force development, maximizing work output or speed. Moderate-load PROM training, common among recreational weight trainers, is unlikely to provide higher movement kinetics

From QTL to causative gene.

<p>Three phases of QTL gene identification are depicted, with the length of time for each phase shown for phenotypes, such as cancer endpoints, that may take several months to a year to assess. a) Linkage analysis. Two inbred mouse or rat strains that differ in cancer risk are intercrossed. Chromosomal segments from the resistant strain (R) are shown in white and those from the susceptible strain (S) are shown in red. After phenotyping 50–200 progeny and genotyping them at approximately 100 markers, linkage analysis reveals two QTLs (<i>QTL1</i> and <i>QTL2</i>) on chromosomes 2 and 4. b) Fine mapping. A congenic line carrying the <i>QTL1</i> chromosomal region from the sensitive strain on the genetic background of the resistant strain is constructed by repeated backcrossing to the resistant strain, with selection for markers from the sensitive strain in the desired interval. The heterozygous congenic is backcrossed to the resistant strain to produce recombinant lines (X1 to X10) that carry various segments of the region from the sensitive strain (shown as red lines). A smaller chromosomal region (Minimal Interval) containing <i>QTL1</i> is inferred from phenotypic analysis of the lines. c) Candidate prioritization and testing. Candidate genes within the minimal interval may be prioritized by several complementary approaches. Comparison of SNP haplotypes for the resistant and sensitive strains (R and S) to those for other strains, such as S2, another sensitive strain with a QTL mapping to the same region as for S, may allow definition of a smaller interval for <i>QTL1</i>. Direct sequence analysis of alleles in the resistant (R) and sensitive (S) strains may identify polymorphisms with potential biological consequences. Differential expression of candidate genes in tumors or appropriate normal tissues from sensitive and resistant recombinant lines may be assessed by transcriptome analysis. Mutations or altered expression of the orthologue of a candidate may be observed in human tumors. Ultimately, a candidate gene may be tested directly for its role in cancer development by modifying the genome of the resistant or sensitive strain through transgenesis or homologous recombination.</p