Antisense Approaches in Analyzing the Functional Role of Proteins in the Central Nervous System - Drugs of Abuse: Neurological Reviews and Protocols - page 360

instead of protein synthesis inhibition in intact cells of tissue. Thus, careful
designing and modulation of antisense molecules are required to minimize
neurotoxicity. In general, successful use of an antisense strategy is not always
guaranteed because of the disadvantages described above. The appropriate
and effective use of this method can be better served by acknowledging and
minimizing its limitations.
In this chapter, an example of the in vivo experiment that was recently
conducted in this laboratory with antisense oligos against metabotropic
glutamate receptor subtype 5 (mGluR5) is presented to illustrate different
perspectives of antisense methodology. The mGluR5 antisense oligonucleotides
that were directly infused into the rat dorsal striatum showed a successful
knockdown of mGluR5 expression in striatal neurons and affected cocaine- and
amphetamine-stimulated behaviors and striatal neuropeptide gene expression.
In addition to the mGluR5 receptor, the following protocol and others (18) are
also largely applicable for knocking down many other surface receptors and
intracellular signaling proteins in striatal neurons either in vivo or in vitro.

2. Materials

1. Male young adult rats averaging 200-225 g at time of surgery.

2. Stereotaxic surgical materials on small animals:

a. Stereotaxic apparatus.

b. Dual-pipe fi beroptic illuminators.

c. Heating pad.

d. Small animal clippers.

e. Syringes (5 mL) with 22-gauge needles.

f. Instruments (e.g., scalpel, scissors, spatula, eye forceps, hemostat, autoclip
wound clips, and antoclip applier).

g. High-speed microdrill and steel burrs (Fine Science Tools).

h. Dental carboxylate cement (Durelon).

i. Screwdriver and stainless steel machine screws (Small Parts).

j. Sterile towel/drape, Povidone-iodine swabsticks and cotton swabs.

k. The rat brain atlas by Paxinos and Watson (19) .

3. Hot glass bead sterilizer.

4. Anesthetic solution (4% chloral hydrate).

5. Intrastriatal infusion system:

a. 24-Gauge stainless steel guide cannula cut 10 mm (Small Parts).

b. 30-Gauge stainless steel wire (stylet) cut 10 mm (Small Parts).

c. 30-Gauge stainless steel cannula (injector) 2.5 mm longer than guide cannula
(Small Parts).

d. Standard PE10 catheter tubing and 10- µ L Hamilton syringe.

e. Microliter syringe pump (Harvard Apparatus).