Molecular and cytological analyses of Deschampsia cespitosa populations from Northern Ontario (Canada).

Deschampsia cespitosa is widely dispersed around the globe, particularly in the northern hemisphere. A high tolerance to adverse environmental conditions allows D. cespitosa to colonize and dominate plots of land that are uninhabitable by other plants. The main objective of the present study was to determine the degree of genetic variation and relatedness among D. cespitosa populations from heavy metal contaminated sites and uncontaminated sites in Northern Ontario, using RAPD markers. Genomic DNA samples from individual plants were analyzed using 35 oligonucleotides of random sequence. Twenty-eight of these primers allowed amplification of random polymorphic DNA (RAPD) loci. Overall, 90% of RAPD bands were polymorphic. Analysis of molecular variance revealed that 72% of the variation could be attributed to individual differences within each of the populations. The within- and among-region variations accounted for 14 and 15% of the total molecular variance, respectively. Population-specific RAPD markers were identified. RAPD markers specific to D. cespitosa were isolated, cloned, and characterized. Cytogenetic analysis revealed a high level of aneuploidy in all the populations from Northern Ontario, with chromosome numbers varying from 2n = 18 to 2n = 26.     

The Tau/A152T mutation,a risk factor for frontotemporal‐spectrum disorders,leads to NR2B receptor‐mediated excitotoxicity

We report on a novel transgenic mouse model expressing human full‐length Tau with the Tau mutation A152T (hTau^AT), a risk factor for FTD‐spectrum disorders including PSP and CBD. Brain neurons reveal pathological Tau conformation, hyperphosphorylation, mis‐sorting, aggregation, neuronal degeneration, and progressive loss, most prominently in area CA3 of the hippocampus. The mossy fiber pathway shows enhanced basal synaptic transmission without changes in short‐ or long‐term plasticity. In organotypic hippocampal slices, extracellular glutamate increases early above control levels, followed by a rise in neurotoxicity. These changes are normalized by inhibiting neurotransmitter release or by blocking voltage‐gated sodium channels. CA3 neurons show elevated intracellular calcium during rest and after activity induction which is sensitive to NR2B antagonizing drugs, demonstrating a pivotal role of extrasynaptic NMDA receptors. Slices show pronounced epileptiform activity and axonal sprouting of mossy fibers. Excitotoxic neuronal death is ameliorated by ceftriaxone, which stimulates astrocytic glutamate uptake via the transporter EAAT2/GLT1. In summary, hTau^AT causes excitotoxicity mediated by NR2B‐containing NMDA receptors due to enhanced extracellular glutamate.     

Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation

The signal produced by fluorescence in situ hybridization (FISH) often is inconsistent among cells and sensitivity is low. Small DNA targets on the chromatin are difficult to detect. We report here an improved nick translation procedure for Texas red and Alexa Fluor 488 direct labeling of FISH probes. Brighter probes can be obtained by adding excess DNA polymerase I. Using such probes, a 30 kb yeast transgene, and the rp1, rp3 and zein multigene clusters were clearly detected.