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Manipulation of gene expression in developing or in mature central nervous systems (CNS) holds a promise for the resolution of many compelling neurobiological questions, including the feasibility of gene therapy to treat diseases of the brain. In this context, a number of viral vectors have been used in recent years to introduce and express genes into the CNS. This article discusses a gene transfer system based on the Herpes Simplex Virus-1 (HSV-1). We describe here the use of non-replicating, non-toxic HSV-1 vector, 8117/43, in a series of studies carried in our joint program. This vector proves further the utility of HSV-1 as a delivery vehicle to a number of distinct sites within the CNS.
The studies were carried out on the mesencephalons of adult guinea pigs. The preparations were made by means of the Golgi technique, as well as the Nissl and Klüver-Barrera methods. Four types of neurons were distinguished in the substantia nigra (SN) of the guinea pig: 1. Bipolar neurons of two kinds: the neurons of the first kind have elongated, fusiform perikarya (25–40 µm), whereas the cells of the second kind have rounded and oval perikarya (15–22 µm). These neurons possess two dendritic trunks which arise from the opposite poles of the cell body and run for a relatively long distance. The bipolar neurons are the most numerous in the pars compacta of SN. 2. Triangular neurons with three primary dendrites arising conically from a perikaryon (20–35 µm). They are the most often observed type of neurons in the pars reticulata of SN. 3. Multipolar neurons with quadrangular or oval perikarya (22–35 µm) and 4–5 dendritic trunks which spread out in all directions. 4. Pear-shaped neurons (perikarya 15–25 µm), which have one or two primary dendritic trunks arising from one pole of the cell body. In all the types of neurons an axon originates either from the dendritic trunk or from the soma and is observed only in its initial segment.
Vulnerability of midbrain dopaminergic (DA) neurons in the weaver mouse was studied at postnatal (P) days 8 and 90, in chosen coronal levels throughout the anteroposterior (AP) extent of the substantia nigra pars compacta (SNc). Wild-type (+/+) and homozygous weaver (wv/wv) mice used were the offspring of pregnant dams injected in several cases with tritiated thymidine on embryonic days 11-15. DA neurons were identified for their tyrosine hydroxylase immunoreactivity. Data reveal that at P8, the frequency of both +/+ and wv/wv late-generated DA cells increases from rostral to caudal SNc. No apparent DA-cell loss was observed at P8 in the mutant genotype, irrespective of the AP level considered. However, throughout the AP, there was a significant reduction in the number of these neurons at any level in 90-day-old weavers. Comparison of P8 and P90 +/+ SNc suggests that cell death is not a major aspect in the developmental regulation of normal DA neurons, although numerical cell depletion in the postnatal development of weaver SNc probably results from the amplification of a basal cell-death process, which affected all the coronal levels studied.
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