Koob Andrew O, Ballantyne Scott, Levesque Aime A, Qureshi Amber A, Congdon Sean
Biology Department, University of Hartford, West Hartford, CT 06117.
Biology Department, University of Wisconsin - River Falls, River Falls, WI 54022.
J Undergrad Neurosci Educ. 2021 Dec 24;20(1):A1-A10. eCollection 2021 Fall.
DNA segments with variable number tandem repeats (VNTR) serve as a model for students to learn DNA extraction and polymerase chain reaction (PCR) techniques in biology laboratory courses from the high school to the graduate level. Because of a growing interest in the neurosciences among undergraduates, we have developed a PCR exercise with a focus on the nervous system and behavior, with the aim of inspiring students from all aspects of the neurosciences to appreciate the central dogma and neurogenetics. DRD4 was a good candidate to provide a lab exercise that would be more engaging than VNTR analysis of a non-coding segment. DRD4 encodes for the dopamine D4 receptor and has been controversially associated with 'novelty seeking' or 'wanderlust' behavior. DRD4 has three common variants of the 48 bp sequence on exon 3, easily differentiated through gel electrophoresis. The 2 repeat (2R), 4 repeat (4R) and 7 repeat (7R) of the 48 bp sequence are the most common alleles. The 7R sequences result in the expressed dopamine D4 receptor having less affinity for dopamine binding, which was proposed to be the reason individuals engage in 'novelty seeking' behavior, to increase dopamine release to facilitate more binding to the receptor. Here we demonstrate an enjoyable and simple two lab sequence to analyze DRD4 genotypes that is appropriate for neuroscience and genetics courses.
具有可变数目串联重复序列(VNTR)的DNA片段可作为模型,供学生在从高中到研究生水平的生物实验室课程中学习DNA提取和聚合酶链反应(PCR)技术。由于本科生对神经科学的兴趣日益浓厚,我们开发了一项以神经系统和行为为重点的PCR实验,旨在激发神经科学各方面的学生理解中心法则和神经遗传学。DRD4是一个很好的选择,可以提供一个比非编码片段的VNTR分析更具吸引力的实验室练习。DRD4编码多巴胺D4受体,并且一直存在争议地与“寻求新奇”或“流浪癖”行为相关联。DRD4的外显子3上有48bp序列的三种常见变体,通过凝胶电泳很容易区分。48bp序列的2重复(2R)、4重复(4R)和7重复(7R)是最常见的等位基因。7R序列导致表达的多巴胺D4受体对多巴胺结合的亲和力较低,这被认为是个体表现出“寻求新奇”行为的原因,以增加多巴胺释放,促进更多地与受体结合。在这里,我们展示了一个有趣且简单的两阶段实验流程,用于分析DRD4基因型,该流程适用于神经科学和遗传学课程。