In JoVE (1)
Other Publications (2)
Articles by Matthew A. Florence in JoVE
Pull-down för calmodulin proteiner Kanwardeep S. Kaleka1, Amber N. Petersen1, Matthew A. Florence1, Nashaat Z. Gerges1 1Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin Calmodulin (CAM) pull-down-analys är ett effektivt sätt för att undersöka samspelet mellan CAM med olika proteiner. Denna metod använder CAM-sepharose pärlor för effektiv och specifik analys av CAM-bindande proteiner. Detta ger ett viktigt verktyg för att utforska CAM signalering i cellulär funktion.
Other articles by Matthew A. Florence on PubMed
Neurogranin Enhances Synaptic Strength Through Its Interaction with Calmodulin The EMBO Journal. Oct, 2009 | Pubmed ID: 19713936 Learning-correlated plasticity at CA1 hippocampal excitatory synapses is dependent on neuronal activity and NMDA receptor (NMDAR) activation. However, the molecular mechanisms that transduce plasticity stimuli to postsynaptic potentiation are poorly understood. Here, we report that neurogranin (Ng), a neuron-specific and postsynaptic protein, enhances postsynaptic sensitivity and increases synaptic strength in an activity- and NMDAR-dependent manner. In addition, Ng-mediated potentiation of synaptic transmission mimics and occludes long-term potentiation (LTP). Expression of Ng mutants that lack the ability to bind to, or dissociate from, calmodulin (CaM) fails to potentiate synaptic transmission, strongly suggesting that regulated Ng-CaM binding is necessary for Ng-mediated potentiation. Moreover, knocking-down Ng blocked LTP induction. Thus, Ng-CaM interaction can provide a mechanistic link between induction and expression of postsynaptic potentiation.
Protective Effect of 20-HETE Inhibition in a Model of Oxygen-glucose Deprivation in Hippocampal Slice Cultures American Journal of Physiology. Heart and Circulatory Physiology. Jan, 2012 | Pubmed ID: 22245774 Recent studies have indicated that inhibitors of the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) may have direct neuroprotective actions since they reduce infarct volume following ischemia reperfusion in the brain without altering blood flow. To explore this possibility, the present study utilized organotypic hippocampal slice cultures subjected to oxygen glucose deprivation (OGD) and re-oxygenation to examine whether 20-HETE is released by organotypic hippocampal slices following OGD and whether it contributes to neuronal death through generation of ROS and activation of caspase-3. The production of 20-HETE increased two-fold following OGD and re-oxygenation. Blockade of the synthesis of 20-HETE with N-hydroxy-N'-(4-butyl-2-methylphenol)-formamidine (HET0016) or its actions with a 20-HETE antagonist, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid (6,15-20-HEDE) reduced cell death as measured by the release of lactate dehydrogenase (LDH) and propidium iodide (PI) uptake. Administration of a 20-HETE mimetic, 20-hydroxyeicosa-5(Z),14(Z)-dienoic acid (5,14-20-HEDE) had the opposite effect and increased injury following OGD. The death of neurons following OGD was associated with an increase in the production of ROS and activation of caspase-3. These effects were attenuated by HET0016 and potentiated following administration of 5,14-20-HEDE. These findings indicate that the production of 20-HETE by hippocampal slices is increased following OGD and that inhibitors of the synthesis or actions of 20-HETE protect neurons from ischemic cell death. The protective effect of the 20-HETE inhibitors is associated with a decrease in superoxide production and activation of caspase-3.