PKH26 is an excellent retrograde and anterograde fluorescent tracer characterized by a small injection site and strong fluorescence emission

The fluorescent dye PKH26, which binds mainly to the cell membrane, has long stability that enables the tracing of PKH26-labeled transplanted cells in host tissue. In the present study, we examined whether this fluorescent dye works as a retrograde or anterograde tracer to label neural networks within the central nervous system of adult and postnatal day 3 (P3) mice. A small injection of the dye into the medullospinal junction resulted in the retrograde labeling of corticospinal tract (CST) neurons in layer V of the sensory-motor cortex both in the adult mice and pups. Injection of the dye into the motor cortex of the P3 pups resulted in the anterograde labeling of CST fibers at a single fiber resolution level, although a similar injection of the dye into the motor cortex of the adult mice failed to stain CST fibers anterogradely. These results suggest that, while PKH26 works as a retrograde or anterograde tracer, anterograde labeling of the adult tracts can not be expected.