Regulation of DNA methylation by ethanol induces tissue plasminogen activator expression in astrocytes

Alcohol exposure affects neuronal plasticity in the adult and developing brain. Astrocytes play a major role in modulating neuronal plasticity and are a target of ethanol. Tissue plasminogen activator (tPA) is involved in modulating neuronal plasticity by degrading the extracellular matrix proteins including fibronectin and laminin and is up‐regulated by ethanol in vivo. In this study we explored the hypothesis that ethanol affects DNA methylation in astrocytes thereby increasing expression and release of tPA. It was found that ethanol increased tPA mRNA levels, an effect mimicked by an inhibitor of DNA methyltransferase (DNMT) activity. Ethanol also increased tPA protein expression and release, and inhibited DNMT activity with a corresponding decrease in DNA methylation levels of the tPA promoter. Furthermore, it was observed that protein levels of DNMT3A, but not DNMT1, were reduced in astrocytes after ethanol exposure. These novel studies show that ethanol inhibits DNA methylation in astrocytes leading to increased tPA expression and release; this effect may be involved in astrocyte‐mediated inhibition of neuronal plasticity by alcohol.

     

GENETIC STRUCTURE VARIABILITY OF KELP ALARIA MARGINATA OVER SPACE AND TIME

The genetic variability of Alaria marginata Postels & Ruprecht was investigated spatially and seasonally using the finger printing technique of amplified fragment length polymorphism (AFLP). Using 206 scoreable bands generated by one primer pair, individual plants that were separated by as little as a few decimeters to> 100 km could be distinguished, and followed an isolation-by-distance model. Genetic similarity ranged from 76% for patches (a few decimeters in diameter), to 71% for individual kelp stands (15 m across) and 67% for a group of stands separated by 185 km. Greater genetic similarity of patches occurred at the wave-sheltered site than at wave-exposed site. The influence of wave to genetic variability and the ability to predict gene flow on small stretch of beach were discussed. In one stand, genetic similarities were markedly different between seasons. This seasonal pattern may be the result of different age groups dominating the sampled stands over time. The genetic structure of A. marginata comprises local scale (patch and within stand) heterogeneity and larger scale (between stands) homogeneity.     

Measuring carbon dynamics in field soils using soil spectral reflectance: prediction of maize root density, soil organic carbon and nitrogen content

This paper reports the development of a proximal sensing technique used to predict maize root density, soil carbon (C) and nitrogen (N) content from the visible and near-infrared (Vis-NIR) spectral reflectance of soil cores. Eighteen soil cores (0?C60?cm depth with a 4.6?cm diameter) were collected from two sites within a field of 90-day-old maize silage; Kairanga silt loam and Kairanga fine sandy loam (Gley Soils). At each site, three replicate soil cores were taken at 0, 15 and 30?cm distance from the row of maize plants (rows were 60?cm apart). Each soil core was sectioned at 5 depths (7.5, 15, 30, 45, and 60?cm) and soil reflectance spectra were acquired from the freshly cut surface at each depth. A 1.5?cm soil slice was taken at each surface to obtain root mass and total soil C and N reference (measured) data. Root densities decreased with depth and distance from plant and were lower in the silt loam, which had the higher total C and N contents. Calibration models, developed using partial least squares regression (PLSR) between the first derivative of soil reflectance and the reference data, were able to predict with moderate accuracy the soil profile root density (r ²?=?0.75; ratio of prediction to deviation [RPD]?=?2.03; root mean square error of cross-validation [RMSECV]?=?1.68?mg/cm³), soil% C (r ²?=?0.86; RPD?=?2.66; RMSECV?=?0.48%) and soil% N (r ²?=?0.81; RPD?=?2.32; RMSECV?=?0.05%) distribution patterns. The important wavelengths chosen by the PLSR model to predict root density were different to those chosen to predict soil C or N. In addition, predicted root densities were not strongly autocorrelated to soil C (r?=?0.60) or N (r?=?0.53) values, indicating that root density can be predicted independently from soil C. This research has identified a potential method for assessing root densities in field soils enabling study of their role in soil organic matter synthesis.     

Cholesterol efflux is differentially regulated in neurons and astrocytes: Implications for brain cholesterol homeostasis

Disruption of cholesterol homeostasis in the central nervous system (CNS) has been associated with neurological, neurodegenerative, and neurodevelopmental disorders. The CNS is a closed system with regard to cholesterol homeostasis, as cholesterol-delivering lipoproteins from the periphery cannot pass the blood–brain-barrier and enter the brain. Different cell types in the brain have different functions in the regulation of cholesterol homeostasis, with astrocytes producing and releasing apolipoprotein E and lipoproteins, and neurons metabolizing cholesterol to 24(S)-hydroxycholesterol. We present evidence that astrocytes and neurons adopt different mechanisms also in regulating cholesterol efflux. We found that in astrocytes cholesterol efflux is induced by both lipid-free apolipoproteins and lipoproteins, while cholesterol removal from neurons is triggered only by lipoproteins. The main pathway by which apolipoproteins induce cholesterol efflux is through ABCA1. By upregulating ABCA1 levels and by inhibiting its activity and silencing its expression, we show that ABCA1 is involved in cholesterol efflux from astrocytes but not from neurons. Furthermore, our results suggest that ABCG1 is involved in cholesterol efflux to apolipoproteins and lipoproteins from astrocytes but not from neurons, while ABCG4, whose expression is much higher in neurons than astrocytes, is involved in cholesterol efflux from neurons but not astrocytes. These results indicate that different mechanisms regulate cholesterol efflux from neurons and astrocytes, reflecting the different roles that these cell types play in brain cholesterol homeostasis. These results are important in understanding cellular targets of therapeutic drugs under development for the treatments of conditions associated with altered cholesterol homeostasis in the CNS.     

Net changes of soil C stocks in two grassland soils 26 months after simulated pasture renovation including biochar addition

The use of deep‐rooting pasture species as a management practice can increase the allocation of plant carbon (C) below ground and enhance C storage. A 2‐year lysimeter trial was set up to compare changes in C stocks of soils under either deep‐ or shallow‐rooting pastures and investigate whether biochar addition below the top 10 cm could promote root growth at depth. For this i) soil ploughing at cultivation was simulated in a silt loam soil and in a sandy soil by inverting the 0 to 10 and 10‐ to 20‐cm‐depth soil layers, and a distinctive biochar (selected for each soil to overcome soil‐specific plant growth limitations) was mixed at 10 Mg ha^?1 in the buried layer, where appropriate and ii) three pasture types with contrasting root systems were grown. In the silt loam, soil inversion resulted in a general loss of C (2.0–8.1 Mg ha^?1), particularly in the buried horizon, under shallow‐rooting pastures only. The addition of a C‐rich biochar (equivalent to 7.6 Mg C ha^?1) to this soil resulted in a net C gain (21–40% over the non‐biochar treatment, P < 0.10) in the buried layer under all pastures; this overcame the loss of C in this horizon under shallow‐rooting pastures. In the sandy soil, all pastures were able to maintain soil C stocks at 10–20 cm depth over time, with minor gains of C (1.6–5.1 Mg ha^?1) for the profile. In this soil, the exposure of a skeletal‐ and nutrient‐depleted soil layer at the surface may have fostered root growth at depth. The addition of a nutrient‐rich biochar (equivalent to 3.6 Mg C ha^?1) to this soil had no apparent effect on C stocks. More research is needed to understand the mechanisms through which soil C stocks at depth are preserved.