A technique is described in which rat pups (5-7 days old) are induced into a state of suspended animation by placing them in the freezing compartment of a refrigerator at -20 degrees C for 8-10 minutes. This induces cessation of respiration and heart beat for 12-15 minutes. Surgical pinealectomy is undertaken during this period. Postoperation the animal is gradually warmed and restored to its dam. The effectiveness of pinealectomy was checked after some 400 days by measurement of plasma dark cycle melatonin levels using radioimmune assay. The method is reliable, robust and the results predictable, no melatonin being detectable 400 days after surgery.
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