Genetic analysis of the role of protein kinase C signaling pathways in behaviors by direct gene transfer with HSV-1 vectors.
A genetic intervention strategy is described to elucidate the specific biochemical pathways in identified types of neurons that underlie behavioral adaptations. This strategy contains three parts: A Herpes simplex virus (HSV-1) vector is used to obtain localized gene transfer, a cell type-specific promoter is used to target expression to a particular type of neuron, and a constitutively active signal transduction enzyme is expressed to alter neuronal physiology. To enable this approach, a constitutively active protein kinase C (PKC) was developed which causes a long-lasting, activation-dependent increase in neurotransmitter release from cultured sympathetic neurons. This genetic intervention strategy was tested using the nigrostriatal system: Microinjection of HSV-1 vectors that contain the tyrosine hydroxylase promoter targeted expression to dopaminergic nigrostriatal neurons. Expression of the constitutively active PKC in a small percentage of nigrostriatal neurons (approximately 0.1-2%) produced a long-term (> or = 1 month) change in apomorphine-induced rotational behavior, the amount of rotational behavior correlated with the number of affected nigrostriatal neurons, and D2-like dopamine receptor levels were elevated in the striatal regions innervated by the affected nigrostriatal neurons. The strengths and limitations of this genetic intervention strategy are discussed.
Geller
A I
AI
Division of Endocrinology, Children's Hospital, Boston, MA 02115, USA.
D000818:Animals; D001058:Apomorphine; D001522:Behavior, Animal / Q000502:physiology*; D003342:Corpus Striatum / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics; D009046:Motor Neurons / Q000378:metabolism; D009415:Nerve Net; D018377:Neurotransmitter Agents / Q000096:biosynthesis* / Q000235:genetics; D011493:Protein Kinase C / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D013239:Stereotyped Behavior / Q000187:drug effects / Q000502:physiology*; D013378:Substantia Nigra / Q000378:metabolism
D018377:Neurotransmitter Agents; D011493:Protein Kinase C; D001058:Apomorphine