Novel Object Recognition in the Rat: A Facile Assay for Cognitive Function

Joanne R. Mathiasen1, Amy DiCamillo1

1 Discovery Research, Cephalon, West Chester, Pennsylvania
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.59
DOI:  10.1002/0471141755.ph0559s49
Online Posting Date:  June, 2010
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Abstract

The rat novel object recognition (NOR) assay is a relatively high-throughput, robust, and sensitive procedure for evaluating compounds for cognition-enhancing activity. For the test, rats are given the opportunity to explore two identical objects for a predetermined period of time. After a delay, the animals are then presented with two objects to explore, one of which is the same as in the first exploration trial, the other a new object. Depending on the length of the delay between the two trials, the rats will either explore the novel object for a greater time period, indicating memory for the familiar object, or will explore the novel and familiar objects for the same amount of time, indicating a lack of recall or loss of memory for the familiar object presented during the initial trial. The protocol described in this unit can be used to evaluate the effects of a compound on the short-term/working memory of adult male rats following a 24-hr inter-trial interval. Curr. Protoc. Pharmacol. 49:5.59.1-5.59.15. © 2010 by John Wiley & Sons, Inc.

Keywords: memory; cognition; novel object

     
 
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Table of Contents

  • Introduction
  • Basic Protocol: Evaluation of Pharmacological Effects on Novel Object Recognition (NOR) in Adult Male Rats
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol: Evaluation of Pharmacological Effects on Novel Object Recognition (NOR) in Adult Male Rats

 Materials
  • Adult Wistar rats (200 to 250 g, ~2 months old; Charles River Laboratories)
  • Rodent diet (e.g., LabDiet 5001, PMI Nutrition International)
  • Test compounds (see recipe)
  • Rat housing cages (36.8-cm length × 30.5-cm width × 19.1-cm height)
  • Bedding material (Alpha Dri, Shepherd Specialty Papers)
  • Task lights (Electrix S202 halogen lamp, 50 W)
  • Large open test box constructed of opaque plastic or acrylic material (50-cm length × 50-cm width × 35-cm height; ViewPoint Life Sciences; see Fig. 5.59.3)
  • Light meter (Traceable standard lux meter; Fisher Scientific, cat. no. 06-662-64)
  • Low-odor permanent marker (e.g., Sharpie blue or black marker)
  • Animal balance accurate to 1 g (Ohaus CS 2000)
  • Familiar and Novel Objects (see Fig. 5.59.3 and Critical Parameters)
  • Reusable adhesive (such as Handi-Tak, Super Glue Corporation)
  • Stopwatches/timers with 1-sec precision (Traceable; VWR, cat. no. 62344-585)
  • 18-G Luer gastric needles (if performing oral administration of dosing solution; Animal Feeding/Intubation Needles, Popper & Sons)
     FigureFigure 5.59.3 The test box with two objects, the tower on the left and the spiral on the right; placed in the back corners.
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Figures

  •  FigureFigure 5.59.1 Novel object recognition assay timeline. Acclimation to the test box is performed without objects for 3 min for each rat individually. Trial 1 (T1; sample) proceeds for 3 min. Following an inter-trial interval (ITI; 24 hr for drug testing), the rat is placed in the test box for Trial 2 (T2; choice; 3 min).
    View Image
  •  FigureFigure 5.59.2 Time course for exploration of novel and familiar objects in the choice trial (Spiral or Tower objects). Mean (±SEM) amount of time (sec) a rat spends exploring a familiar (white bars) versus a novel (gray bars) object separated by either a 15-min, 24-hr, or 48-hr inter-trial interval (ITI). In trials separated by 15 min, but not a 24- or 48-hr ITI, the rat spent significantly more time exploring the novel object relative to the familiar object (*p < 0.05 versus familiar, Tukey's post-hoc test following significant two-way repeated measures ANOVA; n = 8 to 12/group). Significantly increased exploration of the novel object at 15 min implies memory for the familiar object at this time, but not following a 24- or 48-hr inter-trial interval. A 24-hr ITI is typically chosen for drug testing.
    View Image
  •  FigureFigure 5.59.3 The test box with two objects, the tower on the left and the spiral on the right; placed in the back corners.
    View Image
  •  FigureFigure 5.59.4 The test box with four objects in the four corners. This is the arrangement used when the two non-experimental rats are exploring the box and setting the scent in the box and on the objects.
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  •  FigureFigure 5.59.5 Selection of objects can affect the time course for forgetting in the NOR test. Time course for exploration of novel and familiar objects in the choice trial (diet 7-UP can or bottle objects). (A) Mean (±SEM) amount of time (sec) a rat spends exploring a familiar (white bars) versus a novel (gray bars) object separated by a 15-min, 24-hr, or 48-hr inter-trial intervals. Using these objects yielded significant memory at all time points examined (*p < 0.05 versus familiar, Tukey's post-hoc test following significant 2-way repeated measures ANOVA; n = 8 to 12/group). (B) When the data are analyzed as to whether the diet 7-UP can or the bottle is the novel object, it was found there is a stronger memory effect which is retained for up to 48 hr when the familiar object is the diet 7-UP can and the novel object is the bottle. There was also a trend towards greater exploration of the bottle during the sample trial (data not shown). This preference for the bottle may have contributed to the enhanced memory (greater exploration of the bottle when novel) that was observed at the longer time points.
    View Image
  •  FigureFigure 5.59.6 Effects of a cognition enhancing compound on short-term memory in the NOR test at a 24-hr inter-trial interval (Spiral and Tower objects). Rats were administered the test compounds 30 min before Trial 2 (Choice trial). A separate group of control animals were tested following a short 15-min inter-trial interval to demonstrate memory for the familiar object at this time (*p < 0.05 versus familiar, Tukey's post-hoc test following significant two-way repeated measures ANOVA; #p < 0.001 versus familiar, paired t-test; n = 8 to 16/group).
    View Image

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Literature Cited

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