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D Standing

Publications and source records attributed to D Standing.

4 recordsLinked to original sources

Overexpression of hippocampal Ca2+/calmodulin-dependent protein kinase II improves spatial memory.

Hippocampal alpha-calcium/calmodulin-dependent protein kinase II (alphaCaMKII) has been implicated in neuronal plasticity and spatial learning. In the present experiment, an adeno-associated virus (AAV) vector was designed to express alphaCaMKII driven by the U6 promotor. Microinfusion of this vector into the rat hippocampus increased alphaCaMKII immunoreactivity by approximately 73% (Western analysis) and improved performance in a water maze task. Locomotor activity and exploratory behavior in an open field task were not altered by the overexpression of alphaCaMKII. These data support a role for alphaCaMKII in spatial or explicit memory storage. The advantages of viral vectors for manipulating target proteins expression compared with genetically modified mouse models are discussed.

Animals↗

Influence of nematodes on resource utilization by bacteria--an in vitro study.

The positive influence of bacterial feeding nematodes on bacterial mediated processes such as organic matter mineralization and nutrient cycling is widely accepted, but the mechanisms of these interactions are not always apparent. Both transport of bacteria by nematodes, and nutritional effects caused by nematode N excretion are thought to be involved, but their relative importance is not known because of the difficulties in studying these interactions in soil. We developed a simple in vitro assay to study complex nematode/bacterial interactions and used it to conduct a series of experiments to determine the potential influence of nematode movement and nutritional effects on bacterial resource use. The system used bacterial feeding and nonfeeding insect parasitic nematodes, and luminescent bacteria marked with metabolic reporter genes. Both nutritional enhancement of bacterial activity and bacterial transport were observed and we hypothesize that in nature, the relative importance of transport is likely to be greater in bulk soil, whereas nematode excretion may have greater impact in the rhizosphere. In both cases, the ability of nematodes to enhance bacterial resource utilization has implications for soil components of biogeochemical cycling.

Animals↗

In vivo inhibition of hippocampal Ca2+/calmodulin-dependent protein kinase II by RNA interference.

Hippocampal alpha-Ca2+/calmodulin-dependent protein kinase II (alpha-CaMKII) has been implicated in spatial learning, neuronal plasticity, epilepsy, and cerebral ischemia. In the present study, an adeno-associated virus (AAV) vector was designed to express green fluorescent protein (GFP) from the CBA promoter and a small hairpin RNA targeting alpha-CaMKII (AAV-shCAM) driven from the U6 promoter. The AAV-shCAM or control vector was microinfused into the rat hippocampus and behavioral testing conducted 19-26 days following surgery. Expression of the marker gene and alpha-CaMKII was evaluated 31 days following AAV infusion. GFP expression was localized to the hippocampus and extended +/-2 mm rostral and caudal from the injection site. Hippocampal alpha-CaMKII was significantly reduced following AAV-shCAM treatment as demonstrated using immunohistochemical and Western analysis. This suppression of alpha-CaMKII was associated with changes in exploratory behavior (open field task) and impaired place learning (water maze task). These results demonstrate the efficacy of a viral-based delivered shRNA to produce gene suppression in a specific circuit of the brain.

Animals↗

Root border cells take up and release glucose-C.

BACKGROUND AND AIMS: Border cells are released from the root tips of many plant species, and can remain viable in the rhizosphere for 1 week. Whether border cells are capable of controlled glucose exchange with their environment was investigated. METHODS: Border cells were removed from Zea mays L. root tips, and immersed in (14)C-labelled D-glucose. In one experiment, the hexose transport inhibitor, phlorizin, was used to investigate active glucose uptake from a range of glucose concentrations. In another experiment, glucose efflux from border cells was monitored over time. KEY RESULTS: Glucose uptake by the border cells increased with increasing glucose concentration from 0.2 to 20 mm. At 0.2 mm glucose, uptake was mainly active, as evidenced by the approx. 60 % inhibition with phlorizin. At 2 and 20 mm glucose, however, uptake was mainly via diffusion, as phlorizin inhibition was negligible. Glucose efflux increased with time for live border cells in both 2 and 20 mm glucose. There was no clear efflux/time pattern for heat-killed border cells. CONCLUSIONS: Border cells actively take up glucose, and also release it. Under our experimental conditions, glucose uptake and efflux were of similar order of magnitude. In the rhizosphere net glucose exchange will almost certainly depend on local soil conditions.

Biological Transport↗