Regulatory Architecture of Gene Expression Variation in the Threespine Stickleback Gasterosteus aculeatus

Much adaptive evolutionary change is underlain by mutational variation in regions of the genome that regulate gene expression rather than in the coding regions of the genes themselves. An understanding of the role of gene expression variation in facilitating local adaptation will be aided by an understanding of underlying regulatory networks. Here, we characterize the genetic architecture of gene expression variation in the threespine stickleback (Gasterosteus aculeatus), an important model in the study of adaptive evolution. We collected transcriptomic and genomic data from 60 half-sib families using an expression microarray and genotyping-by-sequencing, and located expression quantitative trait loci (eQTL) underlying the variation in gene expression in liver tissue using an interval mapping approach. We identified eQTL for several thousand expression traits. Expression was influenced by polymorphism in both cis- and trans-regulatory regions. Transe-QTL clustered into hotspots. We did not identify master transcriptional regulators in hotspot locations: rather, the presence of hotspots may be driven by complex interactions between multiple transcription factors. One observed hotspot colocated with a QTL recently found to underlie salinity tolerance in the threespine stickleback. However, most other observed hotspots did not colocate with regions of the genome known to be involved in adaptive divergence between marine and freshwater habitats.Peer reviewe

Regulatory Architecture of Gene Expression Variation in the Threespine Stickleback Gasterosteus aculeatus

Much adaptive evolutionary change is underlain by mutational variation in regions of the genome that regulate gene expression rather than in the coding regions of the genes themselves. An understanding of the role of gene expression variation in facilitating local adaptation will be aided by an understanding of underlying regulatory networks. Here, we characterize the genetic architecture of gene expression variation in the threespine stickleback (Gasterosteus aculeatus), an important model in the study of adaptive evolution. We collected transcriptomic and genomic data from 60 half-sib families using an expression microarray and genotyping-by-sequencing, and located expression quantitative trait loci (eQTL) underlying the variation in gene expression in liver tissue using an interval mapping approach. We identified eQTL for several thousand expression traits. Expression was influenced by polymorphism in both cis- and trans-regulatory regions. Transe-QTL clustered into hotspots. We did not identify master transcriptional regulators in hotspot locations: rather, the presence of hotspots may be driven by complex interactions between multiple transcription factors. One observed hotspot colocated with a QTL recently found to underlie salinity tolerance in the threespine stickleback. However, most other observed hotspots did not colocate with regions of the genome known to be involved in adaptive divergence between marine and freshwater habitats

Opposing effects of monomeric and pentameric C-reactive protein on endothelial progenitor cells

C-reactive protein (CRP) has been linked to the pathogenesis of atherosclerosis. The dissociation of native, pentameric (p)CRP to monomeric (m)CRP on the cell membrane of activated platelets has recently been demonstrated. The dissociation of pCRP to mCRP may explain local pro-inflammatory reactions at the site of developing atherosclerotic plaques. As a biomarker, pCRP predicts cardiovascular adverse events and so do reduced levels and function of circulating endothelial progenitor cells (EPCs). We hypothesised that mCRP and pCRP exert a differential effect on EPC function and differentiation. EPCs were treated with mCRP or pCRP for 72 h, respectively. Phenotypical characterisation was done by flow cytometry and immunofluorescence microscopy, while the effect of mCRP and pCRP on gene expression was examined by whole-genome gene expression analysis. The functional capacity of EPCs was determined by colony forming unit (CFU) assay and endothelial tube formation assay. Double staining for acetylated LDL and ulex lectin significantly decreased in cells treated with pCRP. The length of tubuli in a matrigel assay with HUVECs decreased significantly in response to pCRP, but not to mCRP. The number of CFUs increased after pCRP treatment. RNA expression profiling demonstrated that mCRP and pCRP cause highly contradictory gene regulation. Interferon-responsive genes (IFI44L, IFI44, IFI27, IFI 6, MX1, OAS2) were among the highly up-regulated genes after mCRP, but not after pCRP treatment. In conclusion, EPC phenotype, genotype and function were differentially affected by mCRP and pCRP, strongly arguing for differential roles of these two CRP conformations. The up-regulation of interferon-inducible genes in response to mCRP may constitute a mechanism for the local regulation of EPC function

The effect of heat treatment of wood on osteoconductivity

Wood is a natural porous fibre composite, which has some structural similarities to bone. Recently, it has been used as a modelling material in developing synthetic fibre-reinforced composite to be used as load-bearing non-metallic artificial bone material. In this study, the behaviour of wood implanted into bone was studied in vivo in the femur bone of the rabbit. Wood was pre-treated by heat, which altered its chemical composition and structure, as well as the biomechanical properties. In the heat treatment, wood's dimensional stability is enhanced, equilibrium moisture content reduces and the biological durability increases. Cone-shaped implants were manufactured from heat-treated (at 200 and 140 °C) birch wood (Betula pubescens) and from untreated birch. A total of 62 implants were placed in the distal femur of 50 white New Zealand rabbits. The behaviour of the implants was studied at 4, 8 and 20 weeks with histological and histometrical analysis. Osteoconductive contact line and the presence of fibrous tissue and foreign body reaction were determined. The amount of fibrous tissue diminished with time, and the absence of foreign body reaction was found to be in correlation to the amount of heat treatment. Histologically found contact between the implant and the host bone at the interface was significantly more abundant in the 200 °C group (avg. 12.8%) vs. the 140 °C (avg. 2.7%) and the untreated groups (avg. 0.6%). It was observed that the heat treatment significantly modified the biological behaviour of the implanted wood. The changes of the wood by heat treatment showed a positive outcome concerning osteoconductivity of the material. © 2009 Acta Materialia Inc.link_to_subscribed_fulltex

Natural composite of wood as replacement material for ostechondral bone defects

Deciduous wood, birch, pretreated by a technique combining heat and water vapor was applied for the reconstruction of bone defects in the knee joint of rabbits. It was observed that wood showed characteristic properties to be incorporated by the host bone during observation time of 4, 8, and 20 weeks. The natural channel structure of wood served as a porous scaffold, allowing host bone growth as small islets into the wood implants. The other properties of heat-treated wood, such as bioactivity, good handling properties, and sufficient biomechanical properties, might be additional favorable factors for the application of wood as a natural composite material for bone and cartilage repair. At the interface of the surfaces of wood and living bone, bonding occurred. The Chemical Interface Model for bonding bone to wood consists of the reactive ions, such as hydroxyl groups - OH, and covalent bonding as well as hydrogen bonding, which originate from both wood and bone. The bone tissue trauma, with its reactive Ca2+ and PO4 3- ions, proteins, and collagen, available for interaction at ionic and nanolevel, are associated with the complicated chemistry in the cellular response of the early bone healing process. It was concluded that heat-treated wood acted like a porous biomaterial scaffold, allowing ongrowth and ingrowth of bone and cartilage differentiation on its surface, and demonstrating osteoconductive contact, bonding at the interface. © 2007 Wiley Periodicals, Inc.link_to_subscribed_fulltex