摘要
目的:检测血管活性肠肽受体2(Vipr2)敲除小鼠多巴胺(DA)等神经递质含量和视网膜电生理,明确Vipr2敲除对视网膜功能的影响。方法:实验研究。利用RT-PCR检测血管活性肠肽(Vip)、Vipr2在4周龄雄性C57BL/6J小鼠眼球组织中的表达水平。通过CRISPR-Cas9基因编辑技术构建Vipr2敲除(Vipr2-KO)小鼠,然后在4周龄时检测Vipr2-KO和Vipr2野生型(Vipr2-WT)小鼠视网膜中DA等神经递质含量及视网膜电生理功能。Vipr2-KO小鼠与Vipr2-WT小鼠DA等神经递质含量比较采用独立样本t检验,而2种小鼠视网膜电生理数据比较采用双因素重复测量方差分析。结果:Vip和Vipr2 mRNA在小鼠视网膜、脉络膜+视网膜色素细胞、角膜和巩膜中均有表达;在虹膜和晶状体中Vipr2呈低表达,未检测到Vip表达。与Vipr2-WT小鼠相比,Vipr2-KO小鼠表现为近视(t=2.51,P=0.017),视网膜DA、3,4-二羟基苯乙酸(DOPAC)、高香草酸(HVA)含量均明显升高(t=3.42,P=0.001;t=2.15,P=0.037;t=3.27,P=0.002),DOPAC/DA比值无变化;而玻璃体液中DA、DOPAC、DOPAC/DA、HVA、去甲肾上腺素含量差异均无统计学意义。视网膜和玻璃体液中天冬氨酸、谷氨酸、丝氨酸、谷氨酰胺、甘氨酸、牛磺酸和γ氨基丁酸含量在Vipr2-KO小鼠与Vipr2-WT小鼠之间差异均无统计学意义。在暗适应不同光刺激强度下(-3.699、-2.201、-0.699、0.301、0.799 log cd·s/m2),Vipr2-KO小鼠相比Vipr2-WT小鼠,b波振幅明显增加(F=8.65,P=0.015)。结论:Vipr2敲除可引起4周龄小鼠屈光向近视发展,影响视网膜中DA、DOPAC、HVA合成代谢,并引起视网膜电生理功能异常。提示Vipr2敲除可能通过影响视网膜功能参与近视形成,但具体作用机制有待进一步研究。
Objective:To determine if the loss of function of retinal vasoactive intestinal peptide receptor 2(Vipr2)affects dopamine(DA)turnover and retinal electrophysiology behavior.Methods:This is an experimental study.RT-PCR analyzed vasoactive intestinal peptide(Vip)and Vipr2 mRNA expression levels in different ocular tissues of 4-week-old male wildtype C57BL/6J mice.CRISPR-Cas9 gene-editing technology constructed the Vipr2-KO mice.Their refraction,turnover of retinal dopamine,and electroretinograms were measured and compared in these two different groups of 4-week-old Vipr2-KO and Vipr2-WT mice.Statistical analysis of the refraction and DA turnover used the independent sample t-test,and the data of retinal electrophysiology were evaluated with the two-way repeated measures ANOVA between these two different groups of mice.Results:Vip and Vipr2 mRNA levels were highly expressed in the retina,choroid/retinal pigment cells,cornea,and sclera.In the iris and lens,Vipr2 mRNA was expressed at very low levels whereas the Vip mRNA expression level was undetectable.The refraction of 4-week-old Vipr2-KO mice was significantly shifted more towards myopia than that in the Vipr2-WT mice(t=2.51,P=0.017).The contents of DA,3,4-dihydroxyphenylacetate(DOPAC),and homovanillic acid(HVA)were significantly increased more in the 4-week-old Vipr2-KO mice than in the Vipr2-WT mice(t=3.42,P=0.001;t=2.15,P=0.037;t=3.27,P=0.002,respectively).However,the levels of DA,DOPAC,DOPAC/DA,HVA,and norepinephrine in the vitreous humor were not altered between these two different groups.The amino acid contents of aspartic acid,glutamic acid,serine,glutamine,glycine,taurine,andγ-aminobutyric acid were invariant in the retina and vitreous humor of these groups.The b-wave amplitudes under scotopic conditions(-3.699,-2.201,-0.699,0.301,0.799 log cd·s/m2)were higher in the 4-week-old Vipr2-KO mice than in its Vipr2-WT counterpart(F=8.65,P=0.015).Conclusions:Loss of Vipr2 function induces a refractive shift towards myopia at age 4 weeks,and alters both the reti
作者
赵福新
汪琪
王莉
阳凡
郑阳洋
赵晨琛
朱禾
周翔天
瞿佳
Fuxin Zhao;Qi Wang;Li Wang;Fan Yang;Yangyang Zheng;Chenchen Zhao;He Zhu;Xiangtian Zhou;Jia Qu(Eye Hospital,Wenzhou Medical University,Wenzhou 325027,China)
出处
《中华眼视光学与视觉科学杂志》
CAS
CSCD
2022年第7期485-493,共9页
Chinese Journal Of Optometry Ophthalmology And Visual Science
基金
国家自然科学基金(82171096、81570881、81170880)
浙江省自然科学基金(LY22H120001、LY18H120005)。
关键词
Vipr2
Vipr2敲除
多巴胺
高效液相色谱法
视网膜电生理
vasoactive intestinal peptide receptor 2
Vipr2 knockout
dopamine
high-performance liquid chromatography
retinal electrophysiology.
