Anatomical and physiological localization of prelabeled grafts in rat hippocampus.
Dissociated rat fetal hippocampal cells were grafted into normal adult rats. The fetal cells were incubated with one of a number of fluorescent compounds at the time of the dissociation to facilitate identification of the individual grafted cells. The fluorescent labels which were analyzed for this purpose included rhodamine latex microspheres, Cascade blue latex beads, rhodamine-dextran-amine, DiI, and carboxyfluorescein ester. The labeled cells were stereotaxically placed as a suspension into normal rat host hippocampi. The rats were sacrificed 2 to 6 weeks after the grafting for in vitro physiological recordings, and the prelabeled grafts were located using fluorescence optics. During intracellular recordings neurons within the prelabeled grafts were injected with Lucifer yellow to visualize the morphology and integration of neuronal processes within the host. Following the recordings the host slices with the grafts were fixed in 4% paraformaldehyde for anatomical analysis. The ability to prelabel cellular grafts allows subsequent anatomical and physiological analysis of the integration of grafted neurons at the resolution of a single neuron. Such an analysis will improve our understanding of the survival, differentiation, migration, and integration of the grafted neurons and their potential to replace lost function in the lesioned hippocampus.
Pyapali, GK; Turner, DA; Madison, RD
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