Continuous and direct infusion of drug solutions in the brain of awake animals: implementation, strengths and pitfalls.

One of the best strategies for understanding an animal's behavior is to study the function of the brain by experimentally modifying brain chemistry temporarily or on a long-term basis. This can be achieved by direct manipulation of neurochemistry of a targeted brain area with various drugs whose in vitro specificity and sensitivity are known. We assume that an animal's behavior is primarily controlled by the integrated performance of neural networks, rather than the action of a "superstar" single neuron which has narrowly tuned selectivity, in a specified brain region. Therefore, the former must be regulated by a large number of combinations of various transmitter/modulator receptors, hormones, growth factors, and other biochemically identifiable and yet unidentified substances. Under certain conditions, the activation of receptor-bound second messenger systems is thought to cause the enhanced expression of particular genes. Given the wide possibilities in manipulating brain chemistry, which may otherwise result in a variety of consequences, it is crucial to have a dependable means of sustaining the steady-state action of a drug for a sufficiently long time period at a targeted area in the brain of behaving animals. In most cases the continuous application of a drug is necessary to counteract its secondary mitigating effect, which is set in action through negative feedback loops and which in effect reduces the primary action of the drug in use. We have developed a technique to answer this need, using the Alzet osmotic minipump as the source of the continuous infusion force. A drug solution is continuously and directly infused, guided through a chronically implanted cannula, into a targeted area in the brain of behaving animals. The consequences of such an infusion are assessed, during as well as after the infusion, using various types of measurements in behavior, biochemistry, neurophysiology, pharmacology and morphology. The method has been successfully applied, for example, to the study of developmentally regulated neural plasticity in cat visual cortex. A few preconditions should be satisfied for the method to be properly applied to the brains of live animals. Those are: (1) manufacturing a suitable guide system, i.e., cannula-minipump assembly, for the infusion solution; (2) stereotaxic implantation of a cannula-minipump assembly into a selected brain region; and (3) estimating the concentration gradient of the continuously infused solution. This is crucial to assess the specificity and sensitivity of a drug for its assumed effects in vivo.