The following projects are currently available:
Analysis of evoked and spontaneous calcium transients in glutamatergic neurons and Parvalbumin-positive interneurons (cloning, cell culture, live imaging)
Glutamate imaging using the slow sensor for Cav2.1 splice variants to assess decay times (asynchronous release) (cell culture, live imaging)
Transfection and antibody validation for Cav2.1-splice variants-Halo-Tag (cell culture, confocal microscopy)
Analysis of synaptic replacement of endogenous Cav2.1 channels and affinity of Cav2.1 splice variants in the presynapse(cell culture, confocal microscopy)
Impact of individual Cav2.1 splice variants on homeostatic plasticity in neuronal networks (live calcium imaging, confocal microscopy, pharmacology (i.e., TTX, picrotoxin))
Traffic routes of voltage-gated calcium channels, characterization of major pathways to deliver calcium channels to the presynaptic terminal (live imaging, single molecule tracking)
Endogenous labelling of calcium sensor proteins Synaptotagmin 3 and Synaptotagmin 7 combined with their tracking on the cell surface (immunohistochemistry, confocal microscopy, single molecule tracking)
Tracking/counting and manipulating the plasma membrane Ca2+ ATPase (PMCA) in axons (genetic manipulations, immunohistochemistry)
Modulation of calcium signalling by STIM proteins in organotypic hippocampal cultures (STIM knockdown, immunohistochemistry, confocal microscopy, live calcium imaging)
The impact of various Cav2.1 splice variants on mEPSCs and mIPSCs (Cav2.1 knockdown, transfection in neurons, whole-cell patch-clamp)
Cloning and testing of novel IDR-specific Bassoon fragments to study the biophysical properties of the Bassoon protein (cloning, transfection in neurons, fluorescence microscopy, FRAP, droplet fusion analysis)
Analysis of changes in active zone composition after VGCC/scaffolds genetic and acute manipulations (up- or downregulation of Bassoon, immunohistochemistry, confocal microscopy)
Impact of gabapentinoids on synaptic density and stability of connectivity patterns in cultured hippocampal networks (immunohistochemistry, functional imaging, multi-channel recording of neuronal network activity, optogenetic stimulation or suppression of neurons)
Genomic modification of presynaptic targets via CRISPR/Cas9 for imaging (CRISPR/Cas9, neuron subtype-specific expression, immunohistochemistry)
If you are interested or need further details, please contact Prof. Dr. Martin Heine by e-mail.