低聚糖2'-岩藻糖基乳糖改善神经发育和运动能力的综合实验设计

Comprehensive experimental design for the improvement of neurodevelopment and motricity regulated by oligosaccharide 2′-fucosyllactose

秀丽隐杆线虫是评价功能活性物的理想模型之一。该文设计了一个以秀丽隐杆线虫为模型的行为改善和促神经发育活性评价体系,探究了2'-岩藻糖基乳糖(2'-Fucosyllactose,2'-FL)对神经发育和运动能力的影响。实验结果表明,2'-FL延长了线虫的瘫痪寿命,提高了吞咽频率、头摆频率和趋向性,抑制了乙酰胆碱酯酶(AChE)活性,提高了线虫多巴胺、血清素、氨基丁酸和谷氨酸含量,从而改善了线虫神经发育和行为。该实验设计结合了生物科学、食品化学和仪器分析等多门课程知识,综合性较强,能够锻炼学生的实验动手能力和仪器操作能力,提高科学素养及知识迁移和综合应用能力。

[Objective]Establishing a scientific evaluation system is vital to promote scientific research and development in the fields of biological science,food science,biology and medicine,and material chemistry.The purpose of this experiment is to apply the economical,reliable,and efficient model organism Caenorhabditis elegans to the evaluation system for screening and evaluating the neurodevelopmental and behavioral effects of natural active substances.Through this evaluation system,the researcher can systematically assess the specific influence of the bioactive substances on the neurodevelopment function and explain how these substances exert their neuroprotective or stimulative effects by ameliorating motor behavior.Oligosaccharide 2′-fucosyllactose(2′-FL)is a common human milk oligosaccharide(HMO)that accounts for approximately 30%of total HMOs.This oligosaccharide has a significant impact on the health of infants since it can promote the growth of beneficial gut microbiota,such as bifidobacteria.With a deeper understanding of the function of 2′-FL,it is expected to play a greater role in infant nutrition and neurodevelopment.Furthermore,this experimental design also emphasizes the importance of educating and cultivating students.By participating in experimental studies,students can not only deepen their understanding of the biological characteristics of C.elegans,but also improve their professional skills in experimental design and data analysis and enhance their scientific thinking and professional skills in related disciplines,such as biological science,food science,and materials chemistry.[Methods]C.elegans has been widely used in many research fields,such as obesity and metabolism,aging and lifespan,and neurobiology.It has a relatively simple nervous system and no highly specific brain regions.These organisms possess neurotransmitters found in mammals,and their metabolism,vesicle circulation,and other biological processes are highly homologous to those of higher animals.The neuronal lineages and morphology of C.elegans have been fully enumerated,and almost every neuron with precise specificity has a fluorescent reporter gene.Therefore,C.elegans is an excellent experimental model for exploring the neurodevelopmental and behavioral improvement effects of 2′-FL.In this experiment,the paralysis lifespan was first used to screen the effective concentration range of 2′-FL,and then,the pharyngeal pumping rate and head swing frequency of C.elegans were used as indicators to explore the behavioral improvement effect of 2′-FL in C.elegans.Finally,the influence of 2′-FL on neural development was explored by measuring the acetylcholinesterase activity and the level of neurotransmitters in C.elegans.[Results]Oligosaccharide 2′-FL alleviated the paralysis caused by the accumulation of Aβin C.elegans at a concentration of 0.4–2.4 mg/mL.Meanwhile,2′-FL increased the swallowing frequency and head swing frequency of C.elegans,effectively intensified their motor behavior,and enhanced the chemotaxis dysfunction of CL2355.Furthermore,2′-FL significantly raised the proportion of worms with normal phenotypes of cholinergic neurons on days 3,7,and 11 while reducing the proportion of worms with abnormal phenotypes.Oligosaccharide 2′-FL significantly decreased the activity of acetylcholine(AChE)in strain CL4176.Finally,2′-FL boosted the levels of dopamine,serotonin,aminobutyric acid,and glutamate in C.elegans by 132.31%,67.22%,54.68%,and 83.41%,respectively.[Conclusions]Oligosaccharide 2′-FL exhibited a significant improvement in neural development and behavioral function.This experimental design,which was highly comprehensive,not only enhanced the experimental and instrument operation abilities of students but also improved their scientific literacy,knowledge transfer,and comprehensive application ability.