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Updated: Jun 11, 2026
Preparation of Mouse Brain Tissue for Immunoelectron Microscopy
Published on: July 20, 2010
Synaptic and nonsynaptic localization of protocadherin-gammaC5 in the rat brain
Yanfang Li1, David R Serwanski, Celia P Miralles
1Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut 06269-3156, USA.
Insights
Gamma-protocadherin gammaC5 (Pcdh-gammaC5) is a protein found in the brain after birth. It plays a role in synapse formation and neuron-astrocyte interactions during brain development.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Gamma-protocadherins (Pcdh-gammas) are implicated in establishing neuronal connectivity patterns.
- Unlike other Pcdh-gammas expressed during embryonic development, Pcdh-gammaC5 is expressed postnatally, coinciding with peak synaptogenesis.
Purpose of the Study:
- To develop a specific antibody for Pcdh-gammaC5 to investigate its expression and localization in the brain.
- To elucidate the role of Pcdh-gammaC5 in synaptic specificity, maturation, and neuron-astrocyte interactions.
Main Methods:
- Development of a Pcdh-gammaC5 specific antibody.
- Immunohistochemical analysis of Pcdh-gammaC5 expression in various brain regions.
- Culturing hippocampal neurons in the absence of astrocytes.
- Electron microscopic immunocytochemistry in rat brain tissue.
Main Results:
- Pcdh-gammaC5 is highly expressed in specific brain regions including the olfactory bulb, hippocampus, cerebral cortex, and cerebellum.
- Pcdh-gammaC5 is found in both neurons and astrocytes, associated with both GABAergic and glutamatergic synapses.
- Pcdh-gammaC5 localizes both pre- and postsynaptically, extrasynaptically, in cytoplasmic organelles, and in perisynaptic astrocyte processes.
Conclusions:
- Pcdh-gammaC5 likely contributes to synaptic specificity and maturation.
- Pcdh-gammaC5 is involved in neuron-neuron synaptic interactions and neuron-astrocyte communication, including perisynaptic interactions.
Abstract:
It has been proposed that gamma-protocadherins (Pcdh-gammas) are involved in the establishment of specific patterns of neuronal connectivity. Contrary to the other Pcdh-gammas, which are expressed in the embryo, Pcdh-gammaC5 is expressed postnatally in the brain, coinciding with the peak of synaptogenesis. We have developed an antibody specific for Pcdh-gammaC5 to study the expression and localization of Pcdh-gammaC5 in brain. Pcdh-gammaC5 is highly expressed in the olfactory bulb, corpus striatum, dentate gyrus, CA1 region of the hippocampus, layers I and II of the cerebral cortex, and molecular layer of the cerebellum. Pcdh-gammaC5 is expressed in both neurons and astrocytes. In hippocampal neuronal cultures, and in the absence of astrocytes, a significant percentage of synapses, more GABAergic than glutamatergic, have associated Pcdh-gammaC5 clusters. Some GABAergic axons show Pcdh-gammaC5 in the majority of their synapses. Nevertheless, many Pcdh-gammaC5 clusters are not associated with synapses. In the brain, significant numbers of Pcdh-gammaC5 clusters are located at contact points between neurons and astrocytes. Electron microscopic immunocytochemistry of the rat brain shows that 1) Pcdh-gammaC5 is present in some GABAergic and glutamatergic synapses both pre- and postsynaptically; 2) Pcdh-gammaC5 is also extrasynaptically localized in membranes and in cytoplasmic organelles of neurons and astrocytes; and 3) Pcdh-gammaC5 is also localized in perisynaptic astrocyte processes. The results support the notions that 1) Pcdh-gammaC5 plays a role in synaptic specificity and/or synaptic maturation and 2) Pcdh-gammaC5 is involved in neuron-neuron synaptic interactions and in neuron-astrocyte interactions, including perisynaptic neuron-astrocyte interactions.
