Name: an objective and reproducible test of olfactory learning and discrimination in mice
Uploaded: 2018-03-22T13:00:00
Description: Watch this Scientific Journal Video about An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice Video at JoVE.com

This protocol is adapted from previous work within our lab (Huang et al.8). All methods described here have been approved by the Animal Care and Use Committee (ACUC) of Baylor College of Medicine. It is supported by the McNair Medical Institute, NINDS grant R01NS078294 to B.R.A., NIH IDDRC grant U54HD083092, NIDDK grant F30DK112571 to JMP, and NINDS grant F31NS092435 to CKM.

All mice were used under a protocol approved by the Baylor College of Medicine Institutional Animal Care and Use Committee in accordance with NIH standards. Mice used in this protocol were all adult mice (&#62;6 weeks of age) on C57BL6/j background and included both male and female mice.After training/staging tasks, mice are returned to their home cage.
1. Construction and General Rules for Using Operant Learning Box (Figure 1B, C)
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	<li>Assemble a mouse cha...

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The rodent olfactory system provides a unique model to study sensory dependent plasticity. Here we present an olfactory learning paradigm to train mice to associate odorant pairs with either a reward or punishment. Through this learning task, downstream circuit changes can be studied in subsequent experiments (electrophysiology, in vivo neuronal imaging, etc.). Upon completion, mice will learn to perform a simple odor cued task to associate a water reward with one odor, and a timeout punishment with ano...

The mouse OB, where odor information first enters the central nervous system (CNS), provides an excellent model to study experience-dependent structural changes. OB circuity continually integrates adult-born neurons in an activity-dependent manner. Adult-born neuron precursors divide off from progenitors that line the subventricular zone adjacent to the lateral ventricles1. Upon migrating into the OB, these neuronal precursors either survive, differentiate, and integrate as inhibitory granule cells, or undergo apoptosis2. Selection for cell fate is influenced by olfactory activity, including olfactory learning3,4,5,6. After integration, learning-induced synaptic changes occur in granule cells during a two-week critical period7,8. Thus, assays for olfactory learning are useful for examining how experience-dependent plasticity influences structural and functional reorganization of a mature brain circuit.
This protocol offers one approach to olfactory training by using an operant conditioning paradigm. In this task, water-deprived mice are trained to associate one odor (the &#34;Go&#34; odor) with a water reward and another odor (the &#34;No-Go&#34; odor) with a trial timeout punishment. Mice progress through a graded series of training phases over the course of 2-4 weeks. When training is complete, mice respond to the Go or No-Go odor with discrete, corresponding behaviors (seeking a water reward on Go trials and not seeking the water reward on No-Go trials) (Figure 1A). After training is complete, mice can be further challenged with chemically similar odor pairs to test discrimination or become transitioned to studies investigating how olfactory learning alters the structure or function of the OB. Although odor discrimination tasks have been previously described, most rely on subjective measurements such as number of sniffs between two odorants9,10. Furthermore, the need for human scoring of such tasks is also time-intensive. The Go/No-Go olfactory learning task described in this protocol offers an unbiased, direct measurement of odor discrimination and olfactory learning.

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			<td height="40" style="height:40px;width:216px;">Glass vial</td>
			<td style="width:123px;">Qorpak</td>
			<td style="width:344px;">GLC-01016</td>
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			<td height="40" style="height:40px;">Silicon Tubing</td>
			<td>Thermo Scientific</td>
			<td>86000030</td>
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			<td height="40" style="height:40px;">18 gauge needles</td>
			<td>BD</td>
			<td>305196</td>
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			<td height="40" style="height:40px;">1-Butanol</td>
			<td>Sigma Aldrich</td>
			<td>437603</td>
			<td></td>
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			<td height="40" style="height:40px;">Propionic Acid</td>
			<td>Sigma Aldrich</td>
			<td>402907</td>
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			<td height="40" style="height:40px;">Mouse Chamber</td>
			<td>Med Associates</td>
			<td>ENV-307W</td>
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			<td height="40" style="height:40px;">Chamber Floor</td>
			<td>Med Associates</td>
			<td>ENV-307W-GFW</td>
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			<td height="40" style="height:40px;">Water Port</td>
			<td>Med Associates</td>
			<td>ENV-313W</td>
			<td>Need two</td>
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		<tr height="40">
			<td height="40" style="height:40px;">Odor stimulus</td>
			<td>Med Associates</td>
			<td>ENV-275</td>
			<td>Contain 2 valves to gate odor delivery&#160;</td>
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		<tr height="40">
			<td height="40" style="height:40px;">Odor Port</td>
			<td>Med Associates</td>
			<td>ENV-375W-NPP</td>
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			<td height="40" style="height:40px;">USB Interface</td>
			<td>Med Associates</td>
			<td>DIG-703A-USB</td>
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			<td height="40" style="height:40px;">Desktop Computer with Windows 2000, XP, Vista, or 7</td>
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			<td height="40" style="height:40px;">Flow meter</td>
			<td>VWR</td>
			<td>97004-952</td>
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			<td height="40" style="height:40px;">Behavioral software</td>
			<td>Med Associates</td>
			<td>SOF-735</td>
			<td>This software, which runs each training stage, has now been replaced with Med-PC V</td>
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		<tr height="40">
			<td height="40" style="height:40px;">Data Transfer software</td>
			<td>Med Associates</td>
			<td>SOF-731</td>
			<td>This software formats the data to Excel</td>
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			<td height="40" style="height:40px;">Training Software</td>
			<td>Med Associates</td>
			<td>DIG-703A-USB</td>
			<td>This software is used to program each training stage</td>
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			<td height="40" style="height:40px;">Water Valve</td>
			<td>Neptune Research</td>
			<td>225P012-11</td>
			<td>This valve is used to gate the water delivery. Need Two</td>
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		<tr height="40">
			<td height="40" style="height:40px;">Odor Valve</td>
			<td>Neptune Research</td>
			<td>360P012-42</td>
			<td>This valve is used to gate the odor delivery. Need Two</td>
		</tr>
	</tbody>
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an objective and reproducible test of olfactory learning and discrimination in mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Once mice have learned the olfactory learning task, they can now associate novel odor pairs with reward and punishment. These trained mice normally begin with about 50% accuracy on the Go/No-Go task. The percentage correct can be plotted by trial block as a learning curve for novel odor pairs (Figure 3A). Within 10 block trials, which take most mice less than 30 min to perform, mice are able to correctly discriminate between odors with greate...

Watch this Scientific Journal Video about An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice Video at JoVE.com

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice Video (Video) | JoVE

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

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