benzofurans and thionine

The known structural and functional features of the inferior colliculus provided an advantageous substrate for examining the outcome of neural transplantation. In the present study, tissue from the midbrain tectum of Sprague-Dawley rat fetuses was removed at 15, 16 and 19 days of gestation (E15, E16 and E19), incubated as a dissociated cell suspension for 12 to 15 h in a medium containing True Blue for prelabeling, and injected into the central nucleus of the inferior colliculus (CNIC) of 28 normal male adult conspecifics. Viable tectal grafts were found in 57.5% of host animals with 65.2% located in CNIC. Grafts of E16 tissue had a survival rate of 84.2%, which was twice that of E15 or E19 donors, and there was no difference in graft survival rate in hosts sacrificed up to 6 months after the implantation procedure. Neurons prelabeled with True Blue and counterstained with thionin were identified as uniquely green stained cells in brightfield illumination and brilliant opaque cells in darkfield microscopy. Grafts were organized in tightly packed clusters of cells including large rounded polygonal and smaller cells similar in size to normal CNIC large and small multipolar neurons. Cell morphology of the larger ovoid and fusiform grafted neurons most closely resembled that of typical adult CNIC cell populations; other implanted cell types appeared immature even when host post-implant survival was extended. At longer post-implant intervals graft and host cells further intermingled, and both cellular and fibrous bridges were observed at the interface between host and graft tissue. The present work demonstrates the viability and development of homotypic cells grafted into a central auditory structure utilizing a simple method of prelabeling donor tissue with a fluorescent dye.

Topics: Animals; Benzofurans; Coloring Agents; Cytological Techniques; Fetus; Fluorescent Dyes; Gestational Age; Graft Survival; Inferior Colliculi; Male; Microscopy, Fluorescence; Phenothiazines; Rats; Rats, Inbred Strains; Superior Colliculi; Time Factors; Tissue Donors