Double labelling of neural grafts for identification of sites mediating growth in snails

Several parameters were studied in an experiment on intracerebral neural grafts in young snails (Helix aspersa aspersa) in which growth was blocked by removal of the mesocerebrum. The results demonstrate that transplantation of adult mesocerebrum neurons from another subspecies (H aspersa maxima) into the location of the ablated mesocerebrum in the brain of a young juvenile host, leads to functional recovery. In addition, double labelling with charcoal for topographical localisation and fast blue for observation of survival and integration of fluorescent cells within the host brain demonstrated the tolerance to the grafted cells and the successful growth of the chimeric brain. The surviving neurons were mainly located in the median region of the reconstituted brain near the islets of metacerebrum neurons. It is possible that some trophic factors promote growth in the host brain and development of the grafted cells. However, resumption of growth requires the presence of the neurosecretory cells particular to the mesocerebrum which secrete growth hormone. The fast-blue dye, which is still detectable after several months, is useful to track single cells for the study of the functional capacities of the different populations of neurons in the cerebral ganglia and for exploring neuronal replacement strategies in the damaged brain.