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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Traumatic brain injury (TBI) is commonly associated with memory impairment. Here, we present a protocol to assess spatial working memory after TBI via a metric task. A metric test is a useful tool to study spatial working memory impairment after TBI.

Abstract

Impairments to sensory, short-term, and long-term memory are common side effects after traumatic brain injury (TBI). Due to the ethical limitations of human studies, animal models provide suitable alternatives to test treatment methods, and to study the mechanisms and related complications of the condition. Experimental rodent models have historically been the most widely used due to their accessibility, low cost, reproducibility, and validated approaches. A metric test, which tests the ability to recall the placement of two objects at various distances and angles from one another, is a technique to study impairment in spatial working memory (SWM) after TBI. The significant advantages of metric tasks include the possibility of dynamic observation, low cost, reproducibility, relative ease of implementation, and low stress environment. Here, we present a metric test protocol to measure impairment of SWM in adult rats after TBI. This test provides a feasible way to evaluate physiology and pathophysiology of brain function more effectively.

Introduction

The prevalence of neurological deficits such as attention, executive function, and certain memory deficits after moderate traumatic brain injury (TBI) is more than 50 percent1,2,3,4,5,6,7,8. TBI can lead to severe impairments in spatial short-term, long-term, and working memory9. These memory impairments have been observed in rodent models of TBI. Rodent models have enabled the de....

Protocol

The experiments were performed following the recommendations of the Declarations of Helsinki and Tokyo and the Guidelines for the Use of Experimental Animals of the European Community. The experiments were approved by the Animal Care Committee of Ben-Gurion University of the Negev. A protocol timeline is illustrated in Figure 1.

1. Surgical procedures and fluid percussion TBI

  1. Select male and female adult Sprague-Dawley rats, housed at a room temperatur.......

Representative Results

The significance of comparisons between groups was determined using the Mann-Whitney test. Statistical significance of results was considered at P < 0.05, while statistically high relevance was measured at P < 0.01.

The results showed no differences in NSS between all groups before intervention and 28 days after TBI. Each group consisted of 12 female or 12 male rats. The NSS scores obtained 48 h after TBI are presented in Table 1. Rats from the TBI group that showed si.......

Discussion

By specifically targeting the metric spatial information process, this metric test provides a necessary tool toward understanding memory deficiency after TBI. The protocol presented in this paper is a modification of previously described behavioral tasks11. One previously described metric task used two different paradigms, each consisting of three habituation sessions and one testing session. The first paradigm consisted of moving the familiar objects closer together after habituation and the seco.......

Acknowledgements

We thank Professor Olena Severynovska; Maryna Kuscheriava M.Sc; Maksym Kryvonosov M.Sc; Daryna Yakumenko M.Sc; Evgenia Goncharyk M.Sc; and Olha Shapoval, PhD candidate at the Department of Physiology, Faculty of Biology, Ecology, and Medicine, Oles Honchar Dnipro University, Dnipro, Ukraine for their supportive and useful contributions. The data was obtained as part of Dmitry Frank's PhD dissertation.

....

Materials

NameCompanyCatalog NumberComments
2% chlorhexidine in 70% alcohol solutionSIGMA - ALDRICH500 ccFor general antisepsis of the skin in the operatory field
 Bupivacaine 0.1 %
4 boards of different thicknesses (1.5cm, 2.5cm, 5cm and 8.5cm)This is to evaluate neurological defect
4-0 Nylon suture4-00
BottlesTechniplastACBT0262SU150 ml bottles filled with 100 ml of water and 100 ml 1%(w/v) sucrose solution
Bottlses (four) for topological an metric tasksFor objects used two little bottles, first round (height 13.5 cm) and second faceted (height 20 cm) shape and two big faceted bottles, first 9x6 cm (height 21 cm) and second 7x7 cm (height 21 cm).
Diamond Hole Saw Drill 3mm diameterGlass Hole Saw KitOptional. 
Digital Weighing ScaleSIGMA - ALDRICHRs 4,000
Dissecting scissorsSIGMA - ALDRICHZ265969
Ethanol 99.9 % Pharmacy5%-10% solution used to clean equipment and remove odors
EthoVision XT (Video software)Noldus, Wageningen, NetherlandsOptional
Fluid-percussion devicecustom-made at the university workshop   No specific brand is recommended.
Gauze SpongesFisher22-362-178
Gloves (thin laboratory gloves)Optional.
Heater with thermometerHeatingpad-1Model: HEATINGPAD-1/2   No specific brand is recommended.
Horizon-XLMennen Medical Ltd
Isofluran, USP 100%Piramamal Critical Care, IncNDC 66794-017Anesthetic liquid for inhalation
Office 365 ProPlusMicrosoft-Microsoft Office Excel
Olympus BX 40 microscopeOlympus
Operating  forcepsSIGMA - ALDRICH
Operating  ScissorsSIGMA - ALDRICH
PC Computer for USV recording and data analysesIntelIntel® core i5-6500 CPU @ 3.2GHz, 16 GB RAM, 64-bit operating system
Plexiglass boxes linked by a narrow passageTwo transparent 30 cm × 20 cm × 20 cm plexiglass boxes linked by a narrow 15 cm × 15 cm × 60 cm passage
Purina ChowPurina5001Rodent laboratory chow given to rats, mice and hamster is a life-cycle nutrition that has been used in biomedical researc for over 5
Rat cages  (rat home cage or another enclosure)Techniplast2000PNo specific brand is recommended
Scalpel blades 11SIGMA - ALDRICHS2771
SPSSSPSS Inc., Chicago, IL, USA 20 package
Stereotaxic Instrumentcustom-made at the university workshop   No specific brand is recommended
Timing deviceInterval Timer:Timing for recording USV'sOptional. Any timer will do, although it is convenient to use an interval timer if you are tickling multiple rats
Topological and metric tasks deviceSelf made in Ben Gurion University of NegevWhite circular platform 200 cm in diameter and 1 cm thick on table
Video cameraLogitechC920 HD PRO WEBCAMDigital video camera for high definition recording of rat behavior under plus maze test
Windows 10Microsoft

References

  1. Binder, L. M. Persisting symptoms after mild head injury: A review of the postconcussive syndrome. Journal of Clinical and Experimental Neuropsychology. 8 (4), 323-346 (1986).
  2. Binder, L. M. A review of mild h....

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