The projections, and more particularly the ipsilateral projections, from the retina to the dorsal lateral geniculate nucleus (dlGn) and the superior colliculus have been investigated in adult mice of the C57BL/6J strain after rearing in one of four different conditions: 1) after normal visual experience; 2) after unilateral enucleation at birth; 3) in mice with congenital unilateral anophthalmia (in which only one eye develops) 4) in mice with congenital unilateral microphthalmia (in which one eye is of reduced size while the other is normal). In neonatally enucleated and congenitally monocular mice there is an aberrant uncrossed pathway to regions of the dlGn and the superior colliculus which do not normally receive such a projection. This projection is limited in its distribution; in both the neonatally enucleated and the congenitally monocular animals the uncrossed projection does not reach the lateral and dorsal parts of the dlGn and it only innervates the rostral half of the superior colliculus. The density of the uncrossed pathway in these animals is highest in those regions in which the normal uncrossed pathway terminates. In microphthalmic mice the expansion of the uncrossed pathway is less marked than in monocular mice. In the superior colliculus the aberrant uncrossed projections innervate the stratum griseum superficiale where they are often found distributed in small patches. An intertectal crossing of retinal fibers is described from the contralateral superior colliculus to the deprived ipsilateral superior colliculus.

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A single lentivirus vector allowing doxycycline-regulated expression of transgenes in the brain was generated by incorporating the tetracycline (Tet)-dependent regulatory system into the backbone of the vector. Two distinct expression cassettes were inserted upstream and downstream from the central Flap sequence that provides for enhanced transduction of nondividing cells. The first cassette was used to express the transgene under the control of the Tet-dependent minimal cytomegalovirus promoter. The second cassette was employed to express constitutively the Tet-dependent transactivator rtTA2-M2, which activates the Tet-dependent promoter after binding of doxycycline (Tet-on system). Vectors carrying luciferase and tyrosine hydroxylase as the transgene were constructed, tested in astroglia-rich primary cultures, and injected into the striata of rats. The constructs allowed in vitro and in vivo robust expression of the transgene that could be regulated over two orders of magnitude by the addition and withdrawal of doxycycline. The vector may thus be useful for many applications in gene therapy research, including the development of a therapeutic protocol for the treatment of Parkinson's disease based on the restoration of regulated dopamine production.

A new adenoviral vector (Ad-GFAP-GDNF) (Ad-¼ adenovirus, GFAP ¼ glial fibrillary acidic protein, GDNF ¼ glial cell line-derived neurotrophic factor) was constructed in which (i) the E1,E3/E4 regions of Ad5 were deleted and (ii) the GDNF transgene is driven by the GFAP promoter. We verified, in vitro, that the recombinant GDNF was expressed in primary cultures of astrocytes. In vivo, the Ad-GFAP-GDNF was injected into the striatum of rats 1 week before provoking striatal 6-OHDA lesion. After 1 month, the striatal GDNF levels were 37 pg/mg total protein. This quantity was at least 120-fold higher than in nontransduced striatum or after injection of the empty adenoviral vector. At 3 months after viral injection, GDNF expression decreased, whereas the viral DNA remained unchanged. Furthermore, around 70% of the dopaminergic (DA) neurons were protected from degeneration up to 3 months as compared to about 45% in the control groups. In addition, the amphetamine-induced rotational behavior was decreased. The results obtained in this study on DA neuron protection and rotational behavior are similar to those previously reported using vectors with viral promoters. In addition to these results, we established that a high level of GDNF was present in the striatum and that the period of GDNF expression was prolonged after injection of our adenoviral vector.

The transfer of the Glial cell line-derived neurotrophic factor (GDNF) gene to the central nervous system by a recombinant adenoviral vector (Ad) was studied. We constructed the adenovirus vector Ad-NSE-GDNF from which the E1, E3/E4 regions of Ad5 have been deleted and in which the GDNF gene was under the control of a neuron-specific enolase (NSE) promoter. The vector was injected into the striatum of a rat model of Parkinson's disease. We found that (i) the NSE promoter can restrict transgene expression in neurons; (ii) Ad-NSE-GDNF significantly protected dopaminergic (DA) neurons in the substantia nigra (SN) but did not reverse the impairments of amphetamine-induced rotational behavior in lesioned rats.