Exposure to Low-Dose Trichloroethylene Alters Shear Stress Gene Expression and Function in the Developing Chick Heart

Trichloroethylene is an organic solvent used as an industrial degreasing agent. Due to its widespread use and volatile nature, TCE is a common environmental contaminant. Trichloroethylene exposure has been implicated in the etiology of heart defects in human populations and animal models. Recent data suggest misregulation of Ca²⁺ homeostasis in H9c2 cardiomyocyte cell line after TCE exposure. We hypothesized that misregulation of Ca²⁺ homeostasis alters myocyte function and leads to changes in embryonic blood flow. In turn, changes in cardiac flow are known to cause cardiac malformations. To investigate this hypothesis, we dosed developing chick embryos in ovo with environmentally relevant doses of TCE (8 and 800 ppb). RNA was isolated from control and treated embryos at specific times in development for real-time PCR analysis of blood flow markers. Effects were observed on Endothelin-1 (ET-1), Nitric Oxide Synthase-3 (NOS-3) and Krüppel-like Factor 2 (KLF2) expression relative to TCE exposure and consistent with reduced flow. Further, we measured function in the developing heart after TCE exposure by isolating cardiomyocytes and measuring half-width of contraction and sarcomere lengths. These functional data showed a significant increase in half-width of contraction after TCE exposure. These data suggest that perturbation of cardiac function contributes to the etiology of congenital heart defects in TCE-exposed embryos.     

Tuning the molecular giant titin through phosphorylation: Role in health and disease

Titin is a giant multi-functional filament that spans half of the sarcomere. Titin's extensible I-band region functions as a molecular spring that provides passive stiffness to cardiac myocytes. Elevated diastolic stiffness is found in a large fraction of heart failure patients and thus understanding the normal mechanisms and pathophysiology of passive stiffness modulation is clinically important. Here we provide first a brief general background on titin including what is known about titin isoforms and then focus on recently discovered post-translational modifications of titin that alter passive stiffness. We discuss the various kinases that have been shown to phosphorylate titin and address the possible roles of titin phosphorylation in cardiac disease, including heart failure with preserved ejection fraction (HFpEF).     

Perceiving colour at a glimpse: the relevance of where one fixates

We used classification images to examine whether certain parts of a surface are particularly important when judging its colour, such as its centre, its edges, or where one is looking. The scene consisted of a regular pattern of square tiles with random colours from along a short line in colour space. Targets defined by a square array of brighter tiles were presented for 200ms. The colours of the tiles within the target were biased by an amount that led to about 70% of the responses being correct. Subjects fixated a point that fell within the target's lower left quadrant and reported each target's colour. They tended to report the colour of the tiles near the fixation point. The influence of the tiles' colour reversed at the target's border and was weaker outside the target. The colour at the border itself was not particularly important. When coloured tiles were also presented before (and after) target presentation they had an opposite (but weaker) effect, indicating that the change in colour is important. Comparing the influence of tiles outside the target with that of tiles at the position at which the target would soon appear suggests that when judging surface colours during the short "glimpses" between saccades, temporal comparisons can be at least as important as spatial ones. We conclude that eye movements are important for colour vision, both because they determine which part of the surface of interest will be given most weight and because the perceived colour of such a surface also depends on what one looked at last.     

Luminance-color correlation is not used to estimate the color of the illumination

Humans can identify the colors of objects fairly consistently, despite considerable variations in the spectral composition of the illumination. It has been suggested that the correlation between luminance and color within a scene helps to disentangle the influences of illumination and reflectance, because the surfaces that reflect the light of the illuminant well will normally be bright. Because the reliability of the luminance-color correlation as an indicator of the chromaticity of the illuminant depends on the number of surfaces that are considered, we expected the correlation to be determined across large parts of the scene. To examine whether this is so, we compared different scenes with matched luminance and chromaticity, but in which the correlation between luminance and chromaticity was manipulated locally. Our results confirm that there is a bias in perceived color away from the chromaticity of bright surfaces. However, the results show that only the correlation within about 1 degree of the target is relevant. Thus, it is unlikely that the visual system uses the correlation between luminance and color to explicitly determine the chromaticity of the illuminant. Instead, this correlation is presumably implicitly considered in the way that the color contrast at borders is determined.     

Changes in titin and collagen underlie diastolic stiffness diversity of cardiac muscle

Small (N2B) and large (N2BA) cardiac titin isoforms are differentially expressed in a species-specific and heart location-specific manner. To understand how differential expression of titin isoforms may influence passive stiffness of cardiac muscle we investigated the mechanical properties of mouse left ventricular (MLV) wall muscle (expressing predominantly the small titin isoform), bovine left atrial (BLA) wall muscle (predominantly the large isoform), and bovine left ventricular (BLV) wall muscle (expressing small and large isoforms at similar levels). Results indicate that the overall passive muscle stiffness of the muscle types varies nearly ten-fold, with stiffness increasing in the following order: BLA, BLV and MLV. To investigate the basis of the variation in the overall muscle stiffness, the contributions of titin and collagen to muscle stiffness were determined. Results showed that increased muscle stiffness results from increases in both titin- and collagen-based passive stiffness, indicating that titin and collagen change in a co-ordinated fashion. The expression level of the small titin isoform correlates with titin's contribution to overall muscle stiffness, suggesting that differential expression of titin isoforms is an effective means to modulate the filling behavior of the heart.     

Dimerization of the cardiac ankyrin protein CARP: Implications for MARP titin-based signaling

Cardiac ankyrin repeat protein (CARP) and its two close homologs ankrd2 (Arpp) and DARP correspond to a conserved gene family of muscle ankyrin repeat proteins (MARPs). All three genes respond to a variety of stress/strain injury signals with their cytokine-like induction and can associate with the elastic region of titin/connectin. Recently, both CARP and ankrd2 were observed to be elevated in cardiac diseases as well as muscular dystrophies, implicating their joined signaling in muscle diseases. Here we show that CARP in the yeast two-hybrid system (YTH) interacts with itself and desmin. To further verify the YTH data and to investigate possible CARP subunit structure(s), we expressed CARP in E. coli. Expressed CARP has an apparent mobility of about 70 kDa on gel filtration, corresponding to a dimeric species. Yeast two-hybrid experiments using amino- and carboxyterminal deletion clones suggest that CARP, ankrd2, and DARP contain potential coiled-coil dimerization motifs within their unique aminoterminal domains that mediate the formation of homo-dimers. In contrast, we could not detect the formation of hetero-dimers between CARP, ankrd2, and DARP. Therefore, when CARP, ankrd2 and DARP are upregulated in disease/stress states, they are likely to be sorted into distinct structural protein complexes since CARP within the MARP family contains a unique aminoterminal dimerization motif.