Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events Justin Ingram Neuroscience Graduate Student.

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Research interests and techniques: Neuroscience: molecular biology; transgenics, neuroanatomy, physiology, behaviour. Hippocampal function: parvalbuminergic.

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Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events Justin Ingram Neuroscience Graduate Student

1 st step: Generate seizure events in hippocampal slices with 4-AP Ziburkus et al. 2006

The 3 month seizure road block -mothers milk (anti-seizure effect) -diet (low ketone diet) -temperature (34°C-35°C) -pH (7.2) Parameters we changed: -4 AP conc. (0uM-200um) -Na +,K +,Ca 2+,Mg 2+ (incubation/recording chamber) -slice thickness (300um-600um) -agarose support blocks (50um change = no seizure) -age (p21)

Step 2: Record from hippocampal interneurons and pyramidal cells

Creating double transgenic mice which express GFP basket cells (glowing interneuron) Parv/CRE LacZ/GFP Parv/GFP Express GFP on Parvalbumin cells  Glowing Interneurons (Basket Cells)

Step 3: Use nanometer liposomes to measure oxygen fluctuations in pyramidal/basket cells (GFP labeled) Pyramidal Basket Cell (GFP) OLM -horizontal

Fluorescence to measure membrane potential using extracellular dye (Di-8) KCl K+K+ K+K+ K+K+ K+K+ valinomycin K+K+ K+K+ K+K+ K+K+ FL Ratio (UV/Blue-Green) Image plotted with Origin