评估创伤性脑损伤后成年大鼠的显性顺从行为
1Division of Anesthesiology and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences,Ben-Gurion University of the Negev, 2Department of Anesthesiology and Perioperative Medicine,Mayo Clinic, 3Department of Biochemistry and Physiology of the Faculty of Biology and Ecology,Oles Gonchar of the Dnipro National University, 4Department of Ophthalmology, Soroka University Medical Center and the Faculty of Health Sciences,Ben-Gurion University of the Negev
Summary
本协议描述了流体叩击引起的创伤性脑损伤的大鼠模型，然后进行一系列行为测试，以了解主导和顺从行为的发展。将这种创伤性脑损伤模型与特定行为测试结合使用，可以研究脑损伤后的社会障碍。
Abstract
对食物、领地和配偶等资源的竞争显着影响动物物种内部的关系，并通过通常基于支配-顺从关系的社会等级制度进行调解。支配-顺从关系是一个物种个体之间的正常行为模式。创伤性脑损伤是动物对中社会互动障碍和支配-顺从关系重组的常见原因。该协议描述了成年雄性Sprague-Dawley大鼠在诱导创伤性脑损伤后通过诱导后29天至33天之间进行的一系列显性 – 顺从测试与幼稚大鼠相比的顺从行为诱导诱导后29天至33天。显性顺从行为测试显示了脑损伤如何诱导争夺食物的动物的顺从行为。在创伤性脑损伤后，啮齿动物更加顺从，这表明它们在喂食器上花费的时间更少，并且与对照动物相比不太可能首先到达低谷。根据该协议，成年雄性大鼠的创伤性脑损伤后发生顺从行为。
Introduction
当同一物种的成员同时争夺有限的资源时，就会发生种内竞争1.相反，种间竞争发生在两个不同物种的成员之间2。种内竞争分为两种类型，包括干扰（适应）和开发（竞争），其产生取决于争夺的资源类型，例如食物和领土3。
没有支配-顺从关系 （DSR），社会等级制度的存在是不可能的。优势表现为成对动物中的“赢”，从属表现为“输”4。但是，DSR 不仅成对出现，而且以三个或更多为一组出现。1922年，Thorleif Schjelderup-Ebbe描述了家鸡的支配等级制度。主要和从属动物之间的主要区别标志是在喂食器上花费的时间和攻击行为。优势层次结构分为两种形式：线性和非线性5.线性优势涉及两组，A 和 B。在这种传递关系范式6中，A组主导B组，或B组主导A组.当至少存在一个循环关系时，就会发生非线性优势：A主导B，B主导C，C主导A7。
不同物种存在评估支配-顺从行为的模型，包括啮齿动物、鸟类8、非人类灵长类动物9、10、11 和人类12。显性-顺从法在文献中得到了很好的体现，并已被用作评估躁狂和抑郁的模型13，以及抗抑郁药物活性14。该模型已被用于研究成年大鼠母体分离后的早期生活压力15。DSR范式可分为三个模型：显性行为模型13,16的减少，顺从行为模型14的减少和支配模型17的可乐定逆转。
这项研究展示了通过基于食物竞争的任务对DSR的调查。这种方法的优点是易于重现，能够观察和准确分析支配-顺从行为。此外，与类似的行为任务不同，支配-顺从行为任务依赖于食物而不是领土，这使得这种行为任务成本更低、更简单，研究人员不需要经过复杂的训练来执行任务和处理数据。
目前研究的总体目标是证明创伤性脑损伤（TBI）后DSR的发展。TBI 与社交障碍、抑郁和焦虑有关。诱导TBI的模型是一种简单有效的标准模型，涉及使用液体叩诊装置诱导创伤性脑损伤18,19。
Protocol
实验得到了内盖夫本古里安大学动物护理委员会的批准，实验是根据赫尔辛基和东京宣言以及欧洲共同体实验动物护理和使用指南的建议进行的。本研究使用成年雄性Sprague-Dawley大鼠，体重300-350g。将动物饲养在22°C±1°C的室温40%-60%的湿度下，进行明暗循环。 1. 动物制备 随机选择30只成年雄性大鼠，并将它们分为两组：TBI和假大鼠。 随意提供食物?…
Representative Results
神经系统严重程度评分评估使用NSS评估TBI后雄性大鼠的神经功能缺损。将大鼠分为两组：TBI组1组和对照组1组。对照组接受假手术。NSS允许通过积分系统22,23评估运动功能和行为改变;24分表示严重神经功能障碍，0分表示神经功能完整。TBI组和假手术组在手术前1 h的神经功能缺损无统计学意义。与假手术大鼠相比，TBI大鼠术后48小时的神经?…
Discussion
临床研究表明，脑损伤可能会增加患精神疾病的风险26,27。此外，TBI影响社会行为的发展28,29。在该协议中，TBI模型对主导 – 顺从行为的呈现有影响。支配顺从行为表现为在喂食器上花费的时间以及谁先来到喂食器。
除了这里执行的行为任务外，还存在其他用于评估支配-顺从关系的任务?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
所做的工作是德米特里·弗兰克博士论文的一部分。
Materials
| 2% chlorhexidine in 70% alcohol solution | SIGMA – ALDRICH | 500 cc | For general antisepsis of the skin in the operatory field |
| 4 boards of different thicknesses (1.5 cm, 2.5 cm, 5 cm and 8.5 cm) | This is to evaluate neurological defect | ||
| 4-0 Nylon suture | 4-00 | ||
| Bottles | Techniplast | ACBT0262SU | |
| Bupivacaine 0.1 % | |||
| Diamond Hole Saw Drill 3 mm diameter | Glass Hole Saw Kit | Optional. | |
| Digital Weighing Scale | SIGMA – ALDRICH | Rs 4,000 | |
| Dissecting scissors | SIGMA – ALDRICH | Z265969 | |
| Ethanol 99.9 % | Pharmacy | 5%-10% solution used to clean equipment and remove odors | |
| Fluid-percussion device | custom-made at the university workshop | No specific brand is recommended. | |
| Gauze Sponges | Fisher | ||
| Gloves (thin laboratory gloves) | Optional. | ||
| Heater with thermometer | Heatingpad-1 | Model: HEATINGPAD-1/2 | No specific brand is recommended. |
| Horizon-XL | Mennen Medical Ltd | ||
| Isofluran, USP 100% | Piramamal Critical Care, Inc | NDC 66794-017 | Anesthetic liquid for inhalation |
| Logitech Webcam Software | Logitech | 2.51 | Software for video camera |
| Operating forceps | SIGMA – ALDRICH | ||
| Operating Scissors | SIGMA – ALDRICH | ||
| PC Computer for USV recording and data analyses | Intel | Intel core i5-6500 CPU @ 3.2GHz, 16 GB RAM, 64-bit operating system | |
| Plexiglass boxes linked by a narrow passage | Two transparent 30 cm × 20 cm × 20 cm plexiglass boxes linked by a narrow 15 cm × 15 cm × 60 cm passage | ||
| Purina Chow | Purina | 5001 | Rodent laboratory chow given to rats, is a lifecycle nutrition that has been used in biomedical research |
| Rat cages (rat home cage or another enclosure) | Techniplast | 2000P | No specific brand is recommended |
| Scalpel blades 11 | SIGMA – ALDRICH | S2771 | |
| SPSS | SPSS Inc., Chicago, IL, USA | A 20 package | |
| Stereotaxic Instrument | custom-made at the university workshop | No specific brand is recommended | |
| Timing device | Interval Timer:Timing for recording USV's | Optional. Any timer will do, although it is convenient to use an interval timer if you are tickling multiple rats | |
| Video camera | Logitech | C920 HD PRO WEBCAM | Digital video camera for high definition recording of rat behavior under dominant submissive test |
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Cite This Article
Frank, D., Gruenbaum, B. F., Semyonov, M., Binyamin, Y., Severynovska, O., Gal, R., Frenkel, A., Knazer, B., Boyko, M., Zlotnik, A. Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury. J. Vis. Exp. (190), e64548, doi:10.3791/64548 (2022).