Human COL5A1 polymorphisms and quadriceps muscle-tendon mechanical stiffness in vivo.

NEW FINDINGS: What is the central question of the study? Do COL5A1 gene variants, previously reported to have diminished transcript stability, manifest in physiological phenotypes of quadriceps muscle-tendon contractile properties and mechanical stiffness in humans? What is the main finding and its importance? COL5A1 gene variants influence mechanical stiffness, not seeming to affect low-level contractile properties in humans. Functional differences in COL5A1 manifest during moderate- to high-level contractions. Polymorphisms of the collagen type V alpha 1 chain (COL5A1) gene are purported to influence mechanical properties of collagenous tissues. Our purpose was to assess musculotendinous contractile properties of the quadriceps in relationship to the genetic influence of mechanical stiffness. Eighty recreationally active males (aged 19-31 years) were assessed for the presence of three genetic polymorphisms associated with COL5A1 mRNA stability (rs4919510, rs1536482 and rs12722). Genotypes were determined using real-time PCR. Stiffness and contractile properties of the knee musculotendinous complex were assessed by maximal isometric voluntary contractions, ramp isometric voluntary contractions, electrically stimulated contractile events and ultrasonography. All genotype groups were able to activate their knee extensors fully (>97%) as assessed by the interpolated twitch technique and presented no differences in muscle-tendon contractile properties at low submaximal contraction intensities. For the quadriceps muscle-tendon at moderate ramp contractions of 50 and 60% maximal voluntary contraction, the rs12722 CT and TT genotypes had ∼30% greater mean stiffness. The rs1536482 AG and GG genotypes showed a similar trend, but did not achieve statistical significance. Variants of the COL5A1 gene seem to influence quadriceps muscle-tendon stiffness but do not affect low-level contractile properties.

Long-term alterations to DNA methylation as a biomarker of prenatal alcohol exposure: From mouse models to human children with fetal alcohol spectrum disorders.

Rodent models of Fetal Alcohol Spectrum Disorders (FASD) have revealed that prenatal alcohol exposure (PAE) results in differential DNA cytosine methylation in the developing brain. The resulting genome-wide methylation changes are enriched in genes with neurodevelopmental functions. The profile of differential methylation is dynamic and present in some form for life. The methylation changes are transmitted across subsequent mitotic divisions, where they are maintained and further modified over time. More recent follow up has identified a profile of the differential methylation in the buccal swabs of young children born with FASD. While distinct from the profile observed in brain tissue from rodent models, there are similarities. These include changes in genes belonging to a number of neurodevelopmental and behavioral pathways. Specifically, there is increased methylation at the clustered protocadherin genes and deregulation of genomically imprinted genes, even though no single gene is affected in all patients studied to date. These novel results suggest further development of a methylation based strategy could enable early and accurate diagnostics and therapeutics, which have remained a challenge in FASD research. There are two aspects of this challenge that must be addressed in the immediate future: First, the long-term differential methylomics observed in rodent models must be functionally confirmed. Second, the similarities in differential methylation must be further established in humans at a methylomic level and overcome a number of technical limitations. While a cure for FASD is challenging, there is an opportunity for the development of early diagnostics and attenuations towards a higher quality of life.

Changes to histone modifications following prenatal alcohol exposure: An emerging picture.

Epigenetic mechanisms are important for facilitating gene-environment interactions in many disease etiologies, including Fetal Alcohol Spectrum Disorders (FASD). Extensive research into the role of DNA methylation and miRNAs in animal models has illuminated the complex role of these mechanisms in FASD. In contrast, histone modifications have not been as well researched, due in part to being less stable than DNA methylation and less well-characterized in disease. It is now apparent that even changes in transient marks can have profound effects if they alter developmental trajectories. In addition, many histone methylations are now known to be relatively stable and can propagate themselves. As technologies and knowledge have advanced, a small group has investigated the role of histone modifications in FASD. Here, we synthesize the data on the effects of prenatal alcohol exposure (PAE) on histone modifications. Several key points are evident. AS with most alcohol-induced outcomes, timing and dosage differences yield variable effects. Nevertheless, these studies consistently find enrichment of H3K9ac, H3K27me2,3, and H3K9me2, and increased expression of histone acetyltransferases and methyltransferases. The consistency of these alterations may implicate them as key mechanisms underlying FASD. Histone modification changes do not often correlate with gene expression changes, though some important examples exist. Encouragingly, attempts to reproduce specific histone modification changes are very often successful. We comment on possible directions for future studies, focusing on further exploration of current trends, expansion of time-point and dosage regimes, and evaluation of biomarker potential.

Alteration of Gene Expression, DNA Methylation, and Histone Methylation in Free Radical Scavenging Networks in Adult Mouse Hippocampus following Fetal Alcohol Exposure.

The molecular basis of Fetal Alcohol Spectrum Disorders (FASD) is poorly understood; however, epigenetic and gene expression changes have been implicated. We have developed a mouse model of FASD characterized by learning and memory impairment and persistent gene expression changes. Epigenetic marks may maintain expression changes over a mouse's lifetime, an area few have explored. Here, mice were injected with saline or ethanol on postnatal days four and seven. At 70 days of age gene expression microarray, methylated DNA immunoprecipitation microarray, H3K4me3 and H3K27me3 chromatin immunoprecipitation microarray were performed. Following extensive pathway analysis of the affected genes, we identified the top affected gene expression pathway as "Free radical scavenging". We confirmed six of these changes by droplet digital PCR including the caspase Casp3 and Wnt transcription factor Tcf7l2. The top pathway for all methylation-affected genes was "Peroxisome biogenesis"; we confirmed differential DNA methylation in the Acca1 thiolase promoter. Altered methylation and gene expression in oxidative stress pathways in the adult hippocampus suggests a novel interface between epigenetic and oxidative stress mechanisms in FASD.