Ankk1 Loss of Function Disrupts Dopaminergic Pathways in Zebrafish.
Ankk1 Loss of Function Disrupts Dopaminergic Pathways in Zebrafish.
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DOI：
10.3389/fnins.2022.794653
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发表时间：
2022
影响因子：
4.3
通讯作者：
Brennan CH
中科院分区：
文献类型：
--
作者：
Leggieri A;García-González J;Torres-Perez JV;Havelange W;Hosseinian S;Mech AM;Keatinge M;Busch-Nentwich EM;Brennan CH
锚蛋白重复序列和激酶结构域1（ANKK 1）是受体相互作用蛋白丝氨酸/苏氨酸激酶家族的成员，已知参与细胞增殖、分化和转录因子的激活。ANKK 1基因座内的遗传变异被认为在成瘾的易感性中起作用。然而，ANKK 1的作用机制仍然知之甚少。已经表明ANKK 1可能影响多巴胺能通路的发育和/或功能。为了验证这一假设，我们产生了一个CRISPR-Cas9功能缺失的ankk 1斑马鱼系，导致27 bp的插入，破坏了ankk 1序列，引入了一个早期终止密码子。我们发现，ankk 1转录水平显着低于ankk 1突变（ankk 127 ins）鱼相比，他们的野生型（ankk 1 +/+）的兄弟姐妹。在ankk 1 +/+成年斑马鱼脑中，ankk 1蛋白在同皮质、海马、基底外侧杏仁核、中脑和小脑中检测到，类似于哺乳动物的分布模式。相比之下，ankk 1蛋白在ankk 127 ins/27 ins鱼的大脑中减少。定量聚合酶链反应分析显示，在ankk 127 ins的drd 2b mRNA的表达增加，在幼虫和成虫阶段。在ankk 1 +/+成年斑马鱼大脑中，在大脑皮层、小脑、海马和尾状核同源区域检测到了drd 2蛋白，类似于人类的模式。与此相反，drd 2的表达减少，主要是在后脑中发现的ankk 127 ins/27 ins的皮质区域。受精后3天（dpf）幼虫的抗酪氨酸羟化酶免疫染色检测到的细胞体或轴突投射的数量没有差异。行为分析显示，对氨磺必利和阿扑吗啡对运动和惊吓习惯化的影响的敏感性改变，与突触前和突触后受体的广泛丧失一致。Ankk 127 ins突变体表现出降低敏感性的选择性多巴胺受体拮抗剂氨磺必利对运动反应的声音惊吓的影响，并差异敏感的非选择性多巴胺受体激动剂阿扑吗啡对运动和习惯化的影响。总之，我们的研究结果加强了ANKK 1和DRD 2之间的功能关系的假设，支持ANKK 1在多巴胺能通路的维持和/或功能中的作用。需要进一步的工作来解开ANKK 1在不同发育阶段的作用。
Ankyrin repeat and kinase domain containing 1 (ANKK1) is a member of the receptor-interacting protein serine/threonine kinase family, known to be involved in cell proliferation, differentiation and activation of transcription factors. Genetic variation within the ANKK1 locus is suggested to play a role in vulnerability to addictions. However, ANKK1 mechanism of action is still poorly understood. It has been suggested that ANKK1 may affect the development and/or functioning of dopaminergic pathways. To test this hypothesis, we generated a CRISPR-Cas9 loss of function ankk1 zebrafish line causing a 27 bp insertion that disrupts the ankk1 sequence introducing an early stop codon. We found that ankk1 transcript levels were significantly lower in ankk1 mutant (ankk127ins) fish compared to their wild type (ankk1+/+) siblings. In ankk1+/+ adult zebrafish brain, ankk1 protein was detected in isocortex, hippocampus, basolateral amygdala, mesencephalon, and cerebellum, resembling the mammalian distribution pattern. In contrast, ankk1 protein was reduced in the brain of ankk127ins/27ins fish. Quantitative polymerase chain reaction analysis revealed an increase in expression of drd2b mRNA in ankk127ins at both larval and adult stages. In ankk1+/+ adult zebrafish brain, drd2 protein was detected in cerebral cortex, cerebellum, hippocampus, and caudate homolog regions, resembling the pattern in humans. In contrast, drd2 expression was reduced in cortical regions of ankk127ins/27ins being predominantly found in the hindbrain. No differences in the number of cell bodies or axonal projections detected by anti-tyrosine hydroxylase immunostaining on 3 days post fertilization (dpf) larvae were found. Behavioral analysis revealed altered sensitivity to effects of both amisulpride and apomorphine on locomotion and startle habituation, consistent with a broad loss of both pre and post synaptic receptors. Ankk127ins mutants showed reduced sensitivity to the effect of the selective dopamine receptor antagonist amisulpride on locomotor responses to acoustic startle and were differentially sensitive to the effects of the non-selective dopamine agonist apomorphine on both locomotion and habituation. Taken together, our findings strengthen the hypothesis of a functional relationship between ANKK1 and DRD2, supporting a role for ANKK1 in the maintenance and/or functioning of dopaminergic pathways. Further work is needed to disentangle ANKK1’s role at different developmental stages.
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影响因子：
4.5
作者：
Keatinge M;Tsarouchas TM;Munir T;Porter NJ;Larraz J;Gianni D;Tsai HH;Becker CG;Lyons DA;Becker T
通讯作者：
Becker T
影响因子：
3.8
作者：
Glazer, Lilah;Hawkey, Andrew B.;Levin, Edward D.
通讯作者：
Levin, Edward D.
影响因子：
4.6
作者：
Evans JR;Torres-Pérez JV;Miletto Petrazzini ME;Riley R;Brennan CH
通讯作者：
Brennan CH
影响因子：
120.7
作者：
BLUM, K;NOBLE, EP;COHN, JB
通讯作者：
COHN, JB
影响因子：
2.7
作者：
Halberstadt, Adam L.;Geyer, Mark A.
通讯作者：
Geyer, Mark A.