OXYGEN GRADIENTS CAUSE PATTERN ORIENTATION IN DICTYOSTELIUM CELL CLUMPS

We have investigated the formation of the prestalk-prespore pattern in Dictyottelium discoideum. Pattern formation occurs in clumps of Dictyostelium cells embedded in agar under a 100% oxygen atmosphere. Agar embedding allows us to control spatially the environment surrounding the cell clumps. Our results suggest that the ambient oxygen concentration plays a role in controlling the size of the multicellular mass. Further, oxygen gradients established across clumps embedded in agar or held in holes in a plastic barrier cause orientation of the prestalk-prespore pattern such that the anterior prestalk region forms at the highest end of the gradient. The results also indicate that developing cells have the ability to migrate up a gradient of oxygen

Fate and regulation of anterior-like cells in Dictyostelium slugs

A pattern of two tissue types exists in the Dictyostelium slug. Contained within the posterior tissue are anterior-like cells which comprise about 10% of the developing cell mass. For more than 72 hr of slug migration the proportion of these cells is closely regulated. They are randomly distributed along the anterior-posterior axis but about twice as many are localized in the ventral portion of the slug posterior than in the dorsal portion. As the slug begins to form a fruiting body, the anterior-like cells sort out into two groups. One group moves toward the anterior region and one toward the prebasal disc region. In the mature fruiting body the anterior-like cells remain as undifferentiated amoebae at the apex and base of the sorus. Removal of anterior tissue from a slug initiates two events. (1) Some of the anterior-like cells, probably guided by chemotaxis to cyclic AMP, sort out from the posterior tissue. (2) Some prespore cells redifferentiate into anterior-like cells. These events result in the regeneration of a new anterior-posterior pattern after 2 hr and the reestablishment of the original proportions of each cell type by about 8 hr. Furthermore, while the anterior-like cells which lie in slug posteriors remain as amoebae in fruiting bodies, the anterior-like cells which form the anteriors of regenerated slugs subsequently become stalk cells. Thus, it appears that for a cell to differentiate as a stalk cell, it must first be exposed to some form of signal which is present in both the anterior and prebasal disc regions

Cell Sorting daring Pattern Formation in Dictyostelium

Formation of the prestalk-prespore pattern in Dictyostelium was investigated in slugs and submerged clumps of cells. Prestalk and prespore cells were identified by staining with vital dyes, which are shown to be stable cell markers. Dissociated slug cells reaggregate and form slugs that contain a prestalk-prespore pattern indistinguishable from the original pattern. The pattern forms by sorting out of stained prestalk cells from unstained prespore cells. Sorting also occurs in clumps of dissociated slug cells submerged in liquid or agar. A pattern arises in 2 h in which a central core of stained cells is surrounded by a periphery of unstained cells. Sorting appears to be due to differential chemotaxis of stained and unstained cells to cAMP since exogenous cAMP (>10−7 M) reverses the normal direction of sorting-out such that stained cells sort to the periphery of the clumps. Isolated portions of slugs regenerate a new prestalk-prespore pattern. Posterior isolates regenerate a pattern within 2 h due to sorting of a population of vitally stained ‘anterior-like’ cells present in posteriors. Anterior-like cells do not sort in intact slugs due to the influence of a diffusible inhibitor secreted by the anterior region. During posterior regeneration this signal is absent and anterior-like cells rapidly acquire the ability to sort. Anterior isolates regenerate a staining pattern more slowly than posterior isolates by a process that requires conversion of stained prestalk cells to unstained prespore cells. The results suggest that pattern formation in Dictyostelium consists of two processes: establishment of appropriate proportions of two cell types and establishment of the pattern itself by a mechanism of sorting-out

OXYGEN GRADIENTS CAUSE PATTERN ORIENTATION IN DICTYOSTELIUM CELL CLUMPS

We have investigated the formation of the prestalk-prespore pattern in Dictyottelium discoideum. Pattern formation occurs in clumps of Dictyostelium cells embedded in agar under a 100% oxygen atmosphere. Agar embedding allows us to control spatially the environment surrounding the cell clumps. Our results suggest that the ambient oxygen concentration plays a role in controlling the size of the multicellular mass. Further, oxygen gradients established across clumps embedded in agar or held in holes in a plastic barrier cause orientation of the prestalk-prespore pattern such that the anterior prestalk region forms at the highest end of the gradient. The results also indicate that developing cells have the ability to migrate up a gradient of oxygen

Cell Sorting daring Pattern Formation in Dictyostelium

Formation of the prestalk-prespore pattern in Dictyostelium was investigated in slugs and submerged clumps of cells. Prestalk and prespore cells were identified by staining with vital dyes, which are shown to be stable cell markers. Dissociated slug cells reaggregate and form slugs that contain a prestalk-prespore pattern indistinguishable from the original pattern. The pattern forms by sorting out of stained prestalk cells from unstained prespore cells. Sorting also occurs in clumps of dissociated slug cells submerged in liquid or agar. A pattern arises in 2 h in which a central core of stained cells is surrounded by a periphery of unstained cells. Sorting appears to be due to differential chemotaxis of stained and unstained cells to cAMP since exogenous cAMP (>10−7 M) reverses the normal direction of sorting-out such that stained cells sort to the periphery of the clumps. Isolated portions of slugs regenerate a new prestalk-prespore pattern. Posterior isolates regenerate a pattern within 2 h due to sorting of a population of vitally stained ‘anterior-like’ cells present in posteriors. Anterior-like cells do not sort in intact slugs due to the influence of a diffusible inhibitor secreted by the anterior region. During posterior regeneration this signal is absent and anterior-like cells rapidly acquire the ability to sort. Anterior isolates regenerate a staining pattern more slowly than posterior isolates by a process that requires conversion of stained prestalk cells to unstained prespore cells. The results suggest that pattern formation in Dictyostelium consists of two processes: establishment of appropriate proportions of two cell types and establishment of the pattern itself by a mechanism of sorting-out

Genome-wide association studies suggest sex-specific loci associated with abdominal and visceral fat

Background: To identify loci associated with abdominal fat and replicate prior findings, we performed genome-wide association (GWA) studies of abdominal fat traits: subcutaneous adipose tissue (SAT); visceral adipose tissue (VAT); total adipose tissue (TAT) and visceral to subcutaneous adipose tissue ratio (VSR). Subjects and Methods: Sex-combined and sex-stratified analyses were performed on each trait with (TRAIT–BMI) or without (TRAIT) adjustment for body mass index (BMI), and cohort-specific results were combined via a fixed effects meta-analysis. A total of 2513 subjects of European descent were available for the discovery phase. For replication, 2171 European Americans and 772 African Americans were available. Results: A total of 52 single-nucleotide polymorphisms (SNPs) encompassing 7 loci showed suggestive evidence of association (P\u3c1.0 × 10−6) with abdominal fat in the sex-combined analyses. The strongest evidence was found on chromosome 7p14.3 between a SNP near BBS9 gene and VAT (rs12374818; P=1.10 × 10−7), an association that was replicated (P=0.02). For the BMI-adjusted trait, the strongest evidence of association was found between a SNP near CYCSP30 and VAT–BMI (rs10506943; P=2.42 × 10−7). Our sex-specific analyses identified one genome-wide significant (P\u3c5.0 × 10−8) locus for SAT in women with 11 SNPs encompassing the MLLT10, DNAJC1 and EBLN1 genes on chromosome 10p12.31 (P=3.97 × 10–8 to 1.13 × 10−8). The THNSL2 gene previously associated with VAT in women was also replicated (P=0.006). The six gene/loci showing the strongest evidence of association with VAT or VAT-BMI were interrogated for their functional links with obesity and inflammation using the Biograph knowledge-mining software. Genes showing the closest functional links with obesity and inflammation were ADCY8 and KCNK9, respectively. Conclusions: Our results provide evidence for new loci influencing abdominal visceral (BBS9, ADCY8, KCNK9) and subcutaneous (MLLT10/DNAJC1/EBLN1) fat, and confirmed a locus (THNSL2) previously reported to be associated with abdominal fat in women

Physical activity, additional breast cancer events, and mortality among early-stage breast cancer survivors: findings from the WHEL Study

ObjectiveResearch suggests that physical activity is associated with improved breast cancer survival, yet no studies have examined the association between post-diagnosis changes in physical activity and breast cancer outcomes. The aim of this study was to determine whether baseline activity and 1-year change in activity are associated with breast cancer events or mortality.MethodsA total of 2,361 post-treatment breast cancer survivors (Stage I-III) enrolled in a randomized controlled trial of dietary change completed physical activity measures at baseline and one year. Physical activity variables (total, moderate-vigorous, and adherence to guidelines) were calculated for each time point. Median follow-up was 7.1 years. Outcomes were invasive breast cancer events and all-cause mortality.ResultsThose who were most active at baseline had a 53% lower mortality risk compared to the least active women (HR = 0.47; 95% CI: 0.26, 0.84; p = .01). Adherence to activity guidelines was associated with a 35% lower mortality risk (HR = 0.65, 95% CI: 0.47, 0.91; p < .01). Neither baseline nor 1-year change in activity was associated with additional breast cancer events.ConclusionsHigher baseline (post-treatment) physical activity was associated with improved survival. However, change in activity over the following year was not associated with outcomes. These data suggest that long-term physical activity levels are important for breast cancer prognosis

Cell-type specific RNA-Seq reveals novel roles and regulatory programs for terminally differentiated Dictyostelium cells

Abstract Background A major hallmark of multicellular evolution is increasing complexity by the evolution of new specialized cell types. During Dictyostelid evolution novel specialization occurred within taxon group 4. We here aim to retrace the nature and ancestry of the novel “cup” cells by comparing their transcriptome to that of other cell types. Results RNA-Seq was performed on purified mature spore, stalk and cup cells and on vegetative amoebas. Clustering and phylogenetic analyses showed that cup cells were most similar to stalk cells, suggesting that they share a common ancestor. The affinity between cup and stalk cells was also evident from promoter-reporter studies of newly identified cell-type genes, which revealed late expression in cups of many stalk genes. However, GO enrichment analysis reveal the unexpected prominence of GTPase mediated signalling in cup cells, in contrast to enrichment of autophagy and cell wall synthesis related transcripts in stalk cells. Combining the cell type RNA-Seq data with developmental expression profiles revealed complex expression dynamics in each cell type as well as genes exclusively expressed during terminal differentiation. Most notable were nine related hssA-like genes that were highly and exclusively expressed in cup cells. Conclusions This study reveals the unique transcriptomes of the mature cup, stalk and spore cells of D. discoideum and provides insight into the ancestry of cup cells and roles in signalling that were not previously realized. The data presented in this study will serve as an important resource for future studies into the regulation and evolution of cell type specialization