Rationale: The paper uses pre-embedding immuno-EM.
Figure 1 demonstrates the specificity of a newly made antibody against human EAAT4 in this paper (anti-D537).
In figure 6, pre-embedding electron microscopic immunocytochemistry using peroxidase-based labelling shows specific staining in spines, but no staining in axons or in glial processes.
Figure 7: postembedding immunogold EM of EAAT4 demonstrating labelling in membranes of Purkinje cell spines, spine necks, and dendrites.
Figure 8 illustrates that in the vast majority of cases the immunolabelling is seen in contact sites with astroglia and not in contact sites with the postsynaptic membrane, or in the parts of the spine membrane that are in contact with
presynaptic nerve terminals.
Figure 11b - quantification of EAAT4 immunoreactivity along different parts of the Purkinje cell membrane, demonstrating that the majority of particles are found in contact sites to astroglia in dendrite (11b column b) or in spines (11b column f). Experimental description: Animals. Wistar rats from Møllegaard Hansen were kept in the animal
facility at the Institute of Basic Medical Sciences. All handling of animals
was according to European regulations and was under veterinary supervision.
The rats were killed by stunning and decapitation.