Aberrant NOVA1 function disrupts alternative splicing in early stages of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by aberrant alternative splicing (AS). Nuclear loss and cytoplasmic accumulation of the splicing factor TDP-43 in motor neurons (MN) are hallmarks of ALS at late stages of the disease. However, it is unknown if altered AS is present before TDP-43 pathology occurs. Here, we investigate altered AS and its origins in early stages of ALS using human induced pluripotent stem cell-derived motor neurons (MNs) from sporadic and familial ALS patients. We find high levels of the RNA-binding proteins NOVA1, NOVA2, and RBFOX2 in the insoluble protein fractions and observe that AS events in ALS-associated MNs are enriched for binding sites of these proteins. Our study points to an early disrupted function of NOVA1 that drives AS changes in a complex fashion, including events caused by a consistent loss of NOVA1 function. NOVA1 exhibits increased cytoplasmic protein levels in early stage MNs without TDP-43 pathology in ALS postmortem tissue. As nuclear TDP-43 protein level depletes, NOVA1 is reduced. Potential indications for a reduction of NOVA1 also came from mice over-expressing TDP-43 lacking its nuclear localization signal and iPSC-MN stressed with puromycin. This study highlights that additional RBP-RNA perturbations in ALS occur in parallel to TDP-43

Thermal dependence of electrical characteristics of micromachined silica microchannel plates

Micromachined silica microchannel plates (MCPs) under development have a number of advantages over standard glass MCPs and open completely new possibilities in detector technologies. In this article we present the results of our studies on the thermal properties of silica microchannel plates (sMCPs). Similar to standard glass microchanel plates the resistance of silica MCPs was measured to change exponentially with temperature with a negative thermal coefficient of -0.036 per degreesC, somewhat larger than that of standard glass MCPs. The resistance also decreases linearly with the applied voltage, with the voltage coefficient of -3.1x10(-4) V-1. With the knowledge of these two coefficients, our thermal model allows the calculation of the maximum voltage, which can be applied to a given MCP without inducing a thermal runaway. A typical 25 mm diam, 240 mum thick sMCP with 6 mum pores has to have the resistance larger than similar to30 MOmega to operate safely at voltages up to 800 V. With this model we can also calculate the time required for a given silica MCP to reach the point of thermal equilibrium after a voltage increase. We hope that the ongoing efforts on a proper modification of the sMCP semiconducting layer will lead to the production of new MCPs with a small negative or even a positive thermal coefficient, reducing the possibility of thermal runaways of low-resistance MCPs required for high count rate applications. (C) 2004 American Institute of Physics