发布: 2025年09月05日第15卷第17期 DOI: 10.21769/BioProtoc.5428 浏览次数: 248
评审: Xiaochen SunAnonymous reviewer(s)
Abstract
The Morris water maze (MWM) is one of the most widely used procedures to assess hippocampus-dependent spatial learning and memory in rodents. By varying test protocols, researchers can test several different domains of learning and memory. Over multiple testing days, animals learn to swim to a platform hidden just under the water surface by using the spatial relationship between distal cues and the platform. Probe trials, where the platform is rendered unavailable, measure rodents’ spatial bias for the area where the platform was previously located. The ability of researchers to control the availability of the platform “on-demand” offers both practical and methodological advantages. Despite MWM’s prominence in the field of behavioral neuroscience, the high cost of purchasing a commercial MWM package is often prohibitively expensive for many research labs, especially on-demand platforms. Here, we describe a low-cost strategy for a build-your-own MWM that includes a remote-controlled on-demand platform (~530 USD) and tank (~550 USD). It is our hope that disseminating low-cost strategies aimed at expanding access to high-quality research tools at underfunded research institutions will accelerate biomedical discovery and foster further innovation.
Key features
• An on-demand platform allows for seamless testing (e.g., during probe trials, when the platform is usually removed), without interrupting the experiment to adjust/remove the platform.
• The focus here is to offer a step-by-step, economical alternative to otherwise costly, commercial on-demand platforms.
• Additionally, the protocol offers cost-effective ways of assembling the tank, preparing the testing environment, and implementing a testing protocol.
• We include testing strategies specifically developed for aged rats, as these animals are routinely used in our laboratories.
Keywords: Morris water maze (MWM) (Morris水迷宫(MWM))Background
The Morris water maze (MWM) is widely considered a gold standard in behavioral neuroscience for assessing spatial learning and memory in rodents [1]. The basic protocol involves a circular pool filled with opaque water, where a submerged platform is hidden just below the surface. Rats or mice are placed in the water, where they must learn to locate the platform using spatial cues in the environment around the pool. With each learning (acquisition) trial, animals learn the location of the platform and will swim directly to it, demonstrating their spatial learning and memory abilities. The task is used extensively to assess the effects of various genetic, pharmacological, and environmental manipulations on cognitive function.
“Probe tests” provide converging evidence to that acquired by other behavioral measures. In the water maze, probe tests are classically implemented by removing the submerged platform from the tank; the experimenter measures the time the animal spends swimming in the area of the tank where the platform was previously located. Animals that recall the location of the platform spend more of their time near where the platform was previously located. A variant of the original MWM protocol assesses spatial memory over multiple probe trials, not just one. However, removing the platform during these probe trials results in these being extinction trials, since escape is not available. To address this, the probe trial can be modified so that the platform becomes available after a period of time. This procedure maintains response reinforcement in the probe trials as it is for the acquisition trials: go directly to the platform location and remain there. This modification to the protocol limits strategies like circling the wall of the tank until the platform is located, and instead promotes a spatial learning, hippocampus-dependent strategy [2]. Operationally, this requires the height of the platform to be adjustable (“on-demand”) such that it is just under the water surface during acquisition trials, or deep in the tank out of reach from the animal during probe trials [3].
The mechanisms controlling the height of on-demand platforms vary considerably, from cables and/or levers that are not visible to the rat to more technologically advanced methods that include remote-controlled devices. Notable advantages to the latter approach include greater consistency of raising/lowering timing, predictable speed, and minimizing disruption of behavior (e.g., experimenter visibility). However, the cost of such equipment is prohibitively expensive for modestly funded research programs [4]. Here, we developed a do-it-yourself remote-controlled on-demand platform, plus the tank, that costs less than a commercially available counterpart of the platform alone. The primary objective of this report is to disseminate a protocol for constructing a low-cost on-demand platform. However, in addition, we offer an abbreviated account of how we prepared the tank, set up the testing environment, ran a MWM protocol, tracked behavior, and analyzed data. As our aim was not to present a comprehensive MWM how-to, we direct the readers to other resources that, for example, provide more detailed protocols [5–8], detail the many variations of MWM protocols [6], describe its wide ranging applications [9], or offer parallel efforts to lower fiscal barriers to establishing MWM testing [4]. To validate functionality, we implemented our protocol to test whether our setup was sensitive to detect spatial learning and memory deficits among aged F344 rats.
Materials and reagents
Note: Information on vendors, model numbers, and links to purchase equipment are provided when possible. The list of materials used here can also be accessed in Table S1, which also offers information about quantities of materials, prices, and abbreviated notes.
Platform
1. Waterproof linear actuator (Progressive Automations, PA-10-4-450-N-12VDC, https://www.progressiveautomations.com/products/waterproof-linear-actuator-ip68m-for-outdoor-use?variant=31290949697603)
2. Mounting bracket (Progressive Automations, BRK-09, https://www.progressiveautomations.com/products/brk-09)
3. Control box and wireless remote (Progressive Automations, PA-33, https://www.progressiveautomations.com/products/pa-33)
4. Power supply (Progressive Automations, PS-20-12-67, https://www.progressiveautomations.com/products/ps-20-12-67)
5. Polycarbonate plastic sheet (buyplastic.com, PLA-033-029, https://buyplastic.com/products/abrasion-resistant-polycarbonate-ar2-plastic-sheet.html)
6. Flat white spray paint (Home Depot, model: 334021, https://www.homedepot.com/p/Rust-Oleum-Painter-s-Touch-2X-12-oz-Flat-White-General-Purpose-Spray-Paint-334021/307244842)
7. Stainless steel hex nut (Home Depot, model: 800051, https://www.homedepot.com/p/1-4-in-20-Stainless-Steel-Hex-Nut-4-Pack-800051/204274131)
8. Hex flat-head cap screws (Home Depot, model: 44048, https://www.homedepot.com/p/Hillman-1-4-in-x-1-1-2-in-Internal-Hex-Flat-Head-Cap-Screws-8-Pack-44048/204786023)
9. Stainless steel hex bolts (Home Depot, model: RTI2320604, https://www.homedepot.com/p/Robtec-1-4-in-x-2-in-Stainless-Steel-Hex-Bolts-5-Pack-RTI2320604/205710556)
10. Drawer liner (Amazon, model: DSL01-Charcoal-12x10, https://www.amazon.com/gp/product/B0BXSMJK86/ref=ppx_yo_dt_b_search_asin_title?ie=UTF8&th=1)
11. Flat head stainless steel sheet metal screw (Home Depot, model: 814871, https://www.homedepot.com/p/4-x-1-2-in-Phillips-Flat-Head-Stainless-Steel-Sheet-Metal-Screw-8-Pack-814871/204275449?irgwc=1&cm_mmc=afl-ir-2003851-1420157-EdgeBingFlow&clickid=TcOXWgwxLxyKRiEzHBxqD2W6UkCxFvRdRUHC1s0)
Tank
1. Round galvanized stock tank (Tractor Supply Co., SKU: 217713899, https://www.tractorsupply.com/tsc/product/countyline-round-galvanized-stock-tank-6-ft-w-x-2-ft-h-390-gal-capacity)
2. Hose bibb valve (Home Depot, model: 103-054EB, https://www.homedepot.com/p/Everbilt-3-4-MIP-or-1-2-FIP-x-3-4-in-MHT-Forged-Brass-Quarter-Turn-Hose-Bibb-Valve-103-054EB/205822403)
3. Metal filler (Amazon, UPC# 051115904516, https://www.amazon.com/Bondo-90451-Metal-ReinforcedFiller/dp/B010AGXBUO/ref=sr_1_1?dib=eyJ2IjoiMSJ9.35rS9mjvY4pgdC8G7bE1P6VbMBklyrtZka2juNV1F2Zw18XvnwYluhpdqIs1sHuaKOHZe4TxOTH40wQg35dohta2X98x81VGUR4OvwfZdPYhiTzbAAw4xJ7KVsnXpAX0_OVD4WjK8ebva8EfDE1ekJJK)
4. Kitchen and bath sealant (Home Depot, SKU# 1006465750, https://www.homedepot.com/p/DAP-AMP-Advanced-Modified-Polymer-9-oz-White-Kitchen-and-Bath-Sealant-00762/316997383?mtc=SHOPPING-BF-CDP-BNG-D24-024_002_CAULKS-NA-NA-NA-PLALIA-NA-NA-NA-NA-NBR-NA-NA-NA-2024&cm_mmc=SHOPPING-BF-CDP-BNG-D24-024_002_CAULKS-NA-NA-)
5. Rubber stopper, #0, 1-hole (Home Science Tools, SKU: CE-STOP0XA, https://www.homesciencetools.com/product/rubber-stopper-0-1-hole/?_ga=2.102155329.1984278044.1722538286-394657775.1722538286)
Environment
1. Curtain rod (Home Depot, model: AMB144FOHJ07, https://www.homedepot.com/p/Home-Decorators-Collection-72-in-144-in-Telescoping-1-in-Single-Curtain-Rod-in-Matte-Black-AMB144FOHJ07/303673688)
2. Pole sockets (Home Depot, model: 13609, https://www.homedepot.com/p/Everbilt-1-3-8-in-White-Plastic-Pole-Sockets-2-Pack-13609/203170044)
3. Curtain (Home Depot, model: DCV5096BE, https://www.homedepot.com/p/Pro-Space-50-x-96-Outdoor-Patio-Waterproof-Detachable-Sticky-Tab-Top-Porch-Decor-Privacy-Thermal-Insulated-Curtain-Beige-DCV5096BE/315746194)
4. Visual cues (endless options; see note in Procedure)
MWM implementation
1. Permanent marker (Amazon, model: 07888, https://www.amazon.com/Marks-Regular-Permanent-Markers-07888/dp/B000MIJ54M/ref=sr_1_6?dib=eyJ2IjoiMSJ9.K7-yaZSzK6sgBZLtmehNqwvX62jt20HbUxLT-zz-6jmTcD4XF9jGThes6VD6TGCgR8XNwAo92ncQChxJyAhVLnsrKLishBOboFiHiYtj0m6CHTrgwLwgM4aneV3qSa909d4OT5iBaKDjfHR9jy6Bi3Zf)
2. Camera (The Imaging Source, DFK 22AUC03, https://www.oemcameras.com/product/dfk-22auc03-htm/)
3. Lens (The Imaging Source, T2Z 1816 CS, https://dl-gui.theimagingsource.com/en_US/3a27d574-9c67-5e82-a776-6a94ac7bab6c/)
4. USB cable (Amazon, UPC #714133937095, https://www.amazon.com/gp/product/B07HH8KGG2/ref=ppx_yo_dt_b_search_asin_title?ie=UTF8&th=1)
5. Garden hose (Home Depot, Model #82-GHB-25-HD, https://www.homedepot.com/p/GrowGreen-3-4-in-x-25-ft-Expandable-Garden-Hose-82-GHB-25-HD/316267449)
6. Aquarium heater (Amazon, Model #jsmuwkwkP1218, https://www.amazon.com/Orlushy-Submersible-Thermostat-ThermometerFreshwater/dp/B08BPH67FW/ref=sr_1_5?crid=RT4D9N410BO0&dib=eyJ2IjoiMSJ9.riAWZAYvu6eE7LoKWI_p0paziiXTEFZJOxEac8x_LA_S6PUis6ryktXY4iTm2gJTUdFZoL3JekXPn5TVrJs79lyBZ7dsJ0LE_igvxD3DTKCspM-LkJVWZr)
7. Spring clamps (Amazon, Model #SP-04, https://www.amazon.com/EQUIPTZ-Spring-Clamps-inch-Non-Detachable/dp/B0BG8X1KGQ/ref=sr_1_2_sspa?crid=LBMQU6BZGRZG&dib=eyJ2IjoiMSJ9.lvYSPEvVFRsCipKIrvl7uhPyIzXiZSeHIYBxfAqmiP2IUHofGjE_TOYtnn6Flu6N3buMDi1bqxJkHE8LjNkQArKKpivqKXFuEWh9JtyC0xgntU0tTssBjWsAUApst)
8. Wireless presentation clicker (Amazon, model: 910-001350, https://www.amazon.com/Logitech-Professional-Presenter-PresentationWireless/dp/B002GHBUTU/ref=sr_1_3?crid=1BS74AFNU255V&dib=eyJ2IjoiMSJ9.d9kRR8HqvoKhQKJYbBjy4pIIDT8HCd37WJhpRNsb_qh98e0xbueBIgIgti45N6TInqaeb1pEtHiU4okIdSNJZFoc5Yq5cOlIxk54LY8RrRS-c-JKrKQnl)
9. Clamp mount (Amazon, model: RSX-157, https://www.amazon.com/Suptig-Gooseneck-Session-Action-Cameras/dp/B077QDKRC7/ref=sr_1_1?crid=E5FUSOFFWG8Z&dib=eyJ2IjoiMSJ9.TYIvBlMydtU_Q6_iNtWtJH_VeFHIEXvdVGJxQvjpBdHCuILVwVd7c5MmqtXDmMO6cG977M5oR0N4LBqX2LlkQsjNLRW7fD3kH5qLaFX1bwEJSWGSk5DoMm-KFINcx4SCP)
10. Tempera paint (Amazon, model: 214705, https://www.amazon.com/gp/product/B004DJ69MM/ref=ppx_yo_dt_b_search_asin_title?ie=UTF8&th=1)
11. Clamp lamp light (Amazon, model: #2-Pack Clamp Lamp Light, https://www.amazon.com/gp/product/B09J8H4MJ8/ref=ppx_yo_dt_b_search_asin_title?ie=UTF8&th=1)
12. Heat lamp bulbs (Amazon, model: Heat Lamp, https://www.amazon.com/Pyramid-250BR40-Reflector-Medium-Incandescent/dp/B07VWL8D33?th=1)
13. Aquarium fish nets (Amazon, model: QN3, https://www.amazon.com/Penn-Plax-Aquarium-Fish-Net/dp/B0002APXKA/ref=sr_1_1?crid=3OZZF0JT2BPWI&dib=eyJ2IjoiMSJ9.p6FBiO7aWslmlb8T5MWyuA.3s5Z9n2Gd0Ed3SmuEH39PHRx2vpVUFMD52OI1wX6eE&dib_tag=se&keywords=030172230042&qid=1720807319&sprefix=030172230042%2Caps%2)
Equipment
Platform assembly
1. Drill
2. 1/2 in drill bit
3. 1/4 in drill bit
4. 1/16 in drill bit
5. Saw (to cut polycarbonate plastic sheet, e.g., circular saw, jigsaw, band saw)
6. Scissors
Tank assembly
1. Sandpaper
2. Drill
3. 3/4 in drill bit
Software and datasets
1. Behavioral tracking software
Note: While we were fortunate enough to obtain a temporary ANY-maze (Version 7.49) license through an independent loan program, purchasing a standard license of commonly used MWM tracking and analysis software packages (e.g., ANY-maze from Stoelting Co., Ethnovision from Noldus, Smart Video Tracking Software from Panlab, Med Associates, and HVS Image) can be prohibitively expensive for many labs. Fortunately, there is a growing number of open-source options available, including Wintrack [10], Pathfinder [11], MouBeAt [12], Rtrack [13], ezTrack [14], and LabCut [15], though we have not personally tested these potential options.
Procedure
文章信息
稿件历史记录
提交日期: Jun 26, 2025
接收日期: Jul 24, 2025
在线发布日期: Aug 7, 2025
出版日期: Sep 5, 2025
版权信息
© 2025 The Author(s); This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
如何引用
Stinnette, B., Long, J. M. and Myrum, C. (2025). Constructing and Implementing a Low-Cost On-Demand Morris Water Maze Platform. Bio-protocol 15(17): e5428. DOI: 10.21769/BioProtoc.5428.
分类
神经科学 > 行为神经科学 > 学习和记忆
您对这篇实验方法有问题吗?
在此处发布您的问题,我们将邀请本文作者来回答。同时,我们会将您的问题发布到Bio-protocol Exchange,以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述,包括所有有助于他人回答您问题的信息(例如实验过程、条件和相关图像等)。
Share
Bluesky
X
Copy link