1. Protein synthesis inhibition and extinction:
Does cycloheximide produce amnesia for extinction of an odor discrimination in rats?
Alexandra Knoppel, Katherine Janson and Gretchen Hanson Gotthard
Randolph College
Lynchburg, VA 24503
Results
All rats met acquisition and extinction criteria. CHX rats showed amnesia for extinction, while VEH rats did not. More CHX rats (n=7) dug on the test than VEH rats (n=2); χ2=4.337, p=0.037.
Additionally, CHX rats showed amnesia not only for extinction, but for original training as well. CHX rats had preference index scores that did not differ from chance (test 1: t=1.123, p>0.05; test 2: t=2.333, p>0.05). Any VEH rats that dug (n=2) showed perfect preference scores for the correct odor (i.e., 1.0) (see Figure 2).
Introduction
Much research has shown that the administration of protein synthesis inhibitors blocks the formation of new fear memories (e.g., Nader, Schafe, & LeDoux, 2000). In fact, most of the research in this area has focused on the acquisition of fear responses or used tasks that required animals to respond under aversive conditions (e.g., Morris water maze; Meiri & Rosenblum, 1998).
Additionally, most studies examining the effects of blocked protein synthesis have examined acquisition of a completely new response, rather than extinction of an already-established response. A small number of studies have begun to examine the effects of protein synthesis inhibition on extinction (Lattal & Abel, 2001; Suzuki, et al., 2004); however, the results have been mixed and have been conducted with aversive tasks only (e.g., fear conditioning and the water maze).
The present studied used an appetitive odor discrimination digging task (Bunsey & Eichenbaum, 1996) to examine the effects of a protein synthesis inhibitor (cycloheximide - CHX) on extinction in rats. Considering extinction is similar to acquisition in that it also involves new learning, it was hypothesized that protein synthesis inhibition would block extinction and produce continued high levels of responding during testing.
Figure 1
Shaping (Day 1)
One cup of unscented sand (two trials)
Figure 2
Shaping (Day 2)
Two cups of unscented sand (two trials)
Shaping (Day 3)
Two cups of unscented sand (six trials)
Discussion
Rats learned to dig in the correct odor (acquisition) and to stop digging in the correct odor (extinction).
Administration of a protein synthesis inhibitor (CHX) during extinction blocked memory for extinction and for original training.
Results suggest that protein synthesis may be critical for original learning and subsequent extinction learning in an appetitive odor discrimination task.
Training (Day 4)
Trained on one odor discrimination (at least six trials with 75% correct total)
Method
Subjects
Ninety-day old, male Long-Evans rats (N=19) were reduced to and maintained at 85% of their free-feeding weights one week prior to and during experimentation. Water was available ad libitum. Rats were maintained on a 12-hour light/dark cycle.
Apparatus
All shaping, training, and testing took place in the rats’ home cages. Plastic Nalgene® cups (125 ml size) were used for the odor discriminations and were mounted using Velcro® onto rectangular Plexiglas® bases. Odor discriminations were created by mixing play sand (148.5 grams) with different dried spices (i.e., 1.5 grams of cocoa or cinnamon).
Procedure
Rats were shaped to dig in unscented cups of sand for three days prior to training (10 shaping trials total – see Figure 1 for shaping, training, and testing procedure). Rats received one day of training during which they learned one odor discrimination to 75% correct. One day following training, rats received at least two extinction trials for the odor discrimination with a 1 mg/kg CHX injection (n=10) or a vehicle (VEH) injection (n=9). Twenty-four hours later, rats received two test trials.
Extinction (Day 5)
Extinction trials for the odor discrimination (at least two trials and no digging for 30 consecutive seconds)
References
Bunsey, M. & Eichenbaum, H. (1996). Conservation of hippocampal memory function in rats and humans. Nature, 379,
Lattal, K. M. & Abel, T. (2001). Different requirements for protein synthesis in acquisition and extinction of spatial preferences and context-evoked fear. The Journal of Neuroscience, 21(15),
Meiri, N. & Rosenblum, K. (1998). Lateral ventricle injection of the protein synthesis inhibitor anisomycin impairs long-term memory in a spatial memory task. Brain Research, 789,
Nader, K., Schafe, G. E., & LeDoux, J. E. (2000). Fear memories require protein synthesis in the amygdala for reconsolidation after retrieval. Nature, 406,
Suzuki, A., Josselyn, S. A., Frankland, P. W., Masushige, S., Silva, A. J., & Kida, S. (2004). Memory reconsolidation and extinction have distinct temporal and biochemical signatures. The Journal of Neuroscience, 24(20),
Testing (Day 6)
Two test trials for odor discrimination