Correlative Microscopy of Cerebellar Molecular Layer Postsynaptic Proteins and Postsynaptic Receptors | Biomed Grid

Correlative microscopy of cerebellar molecular layer allowed us to image the localization of postsynaptic receptors using a comparative and intermicroscopy study applying transmission electron microscopy (TEM), freeze-etching and direct replicas for transmission electron microscopy (FFTEM), field emission scanning electron microscopy (FESEM), and confocal laser scanning microscopy (CLSM) using immunohistochemistry of Synapsin-I and PSD-95, immunohistochemistry of Ca2+/calmodulin dependent protein kinase II alpha and immunohistochemistry GluR1 subunits of AMPA receptors alpha. These techniques showed the images of postsynaptic densities of parallel fibers-Purkinje dendritic ramifications containing postsynaptic proteins and glutamate postsynaptic receptors. The freeze-etching replica method for TEM showed the three-dimensional structure and intramembrane morphology of postsynaptic ellipsoidal and spheroid intramembrane particles. Confocal laser scanning microscopy using Synapsin-I (Syn-I) and PSD-95, GluR1 subunits of ionotropic glutamate receptors, and Ca2+/calmodulin dependent protein kinase II alpha contribute to image the precise localization of postsynaptic receptors. The high resolution FESEM showed 25-50nm globular subunits at the spine postsynaptic density corresponding to the localization of postsynaptic proteins and/or postsynaptic glutamate receptors