Using GADlacZ transgenic mice as a marker system for homotopic transplantation

Abstract

Olfactory bulb (OB) transplantation is a well characterized model that has been widely used for studying neuronal plasticity and regeneration [G. Sekerková, Z. Katarova, E. Mugnaini, F. Joó, J.R. Wolff, S. Prodan, G. Szabó, Intrinsically labeled relay neurons of homotopic olfactory bulb transplants establish proper afferent and afferent synaptic connections with host neurons, Neuroscience, 80 (1997) 973–979 [10]; G. Sekerková, Z. Malatová, J. Orendáčová, T. Žigová, Transplantation of dorsal root ganglion into the olfactory bulb of neonatal rats: a histochemical study, Restor. Neurol. Neurosci., 6 (1993) 1–8 [11]; E. Račeková, I. Vanický, T. Žigová, Correlation of functional alteration with lesion extent after olfactory bulbectomy in rats, Int. J. Neurosci., 79 (1994) 13–20 [12]; T. Žigová, P.P.C. Graziadei, A.G. Monti Graziadei, Olfactory bulb transplantation into the olfactory bulb of neonatal rats: an autoradiographic study, Brain Res., 539 (1991) 51–58 [13]]. In previous studies, the OB grafts have been routinely labeled by tritiated thymidine [S.M. Onifer, L.A. White, S.R. Whittemore, V.R. Holets, In vitro labelling strategies for identifying primary neural tissue and neuronal cell line after transplantation in the CNS, Cell Transplant., 2 (1993) 131–149 [7]; [13]] allowing distinction of graft from the surrounding tissue by the presence of silver grains over the cell nuclei of the transplant. However, this approach has some disadvantages, namely: partial or insufficient labeling of a defined neuronal subclasses due to the length of the period of their generation, variation in the number of labeled cells due to differences in the gestation stage between individual embryos at the time of i.p. injection of tritiated thymidine, inability to follow the dendritic arborization and axonal outgrowth of the transplanted neurons or to detect directly their actual synaptic contacts, and finally, the need to work with radioactive isotopes. In this paper, we describe an alternative approach, in which the donor OBs in a homotopic OB transplantation were derived from transgenic mice carrying the bacterial gene lacZ under control from the regulatory region of GAD67 gene. In these mice, β-galactosidase (β-gal), encoded by lacZ is stably, ectopically expressed in the vast majority of mitral/tufted (M/T) cells of the OB and served as their intrinsic cellular marker in the OB transplant. By using a simple histochemical reaction for β-gal or immunocytochemistry with anti-β-gal antibody, we could detect the cell bodies and processes of the donor M/T cells and their synaptic contacts with host neurons after long-term survival using both light and electron microscopy. Given the great number of existing transgenic mouse lines that express in the nervous system, this approach may have an even wider application in neural transplantation. Themes: Development and regeneration Topics: Transplantation