Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform.
Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform.
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DOI：
10.3389/fcell.2020.586296
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发表时间：
2020
影响因子：
5.5
通讯作者：
Dennis MY
中科院分区：
文献类型：
--
作者：
Colón-Rodríguez A;Uribe-Salazar JM;Weyenberg KB;Sriram A;Quezada A;Kaya G;Jao E;Radke B;Lein PJ;Dennis MY
近年来，斑马鱼已成为研究人类特征和疾病的常用模型。与其他脊椎动物模型相比，它们的小尺寸，高繁殖力和快速发育允许更多的高通量实验。考虑到斑马鱼与人类共享>70%的基因同源物，并且它们的基因组可以使用高效的CRISPR方法轻松编辑，我们现在能够快速产生影响几乎任何感兴趣基因的突变。不幸的是，我们的能力表型突变幼虫没有跟上步伐。为了应对这一挑战，我们开发了一种方案，该方案以自动化和并行的方式从个体斑马鱼幼虫获得多个表型测量值，包括形态学特征（即，身体长度、眼睛面积和头部尺寸）和运动/行为。通过在各种条件下测定野生型斑马鱼，我们确定了避免显著发育缺陷或物理损伤的最佳参数;这些参数包括在两个时间点[受精后3天（DPF）和5 DPF]对幼虫进行形态学成像，并在5 DPF对行为进行运动跟踪。作为原理证明，我们在两种新的CRISPR生成的突变斑马鱼品系上测试了我们的方法，这两种斑马鱼品系携带syngap 1b和slc 7a 5的预测空等位基因，这两种基因与自闭症谱系障碍、智力残疾和癫痫有关。使用我们优化的高通量表型分析方案，我们概括了先前发表的这些候选基因的小鼠和斑马鱼模型的结果。总之，我们描述了一个快速的并行管道，以强大和一致的方式表征个体幼虫的形态和行为特征，从而提高我们更好地识别人类特征和疾病中重要基因的能力。
In recent years, zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with humans and their genomes can be readily edited using highly efficient CRISPR methods, we are now able to rapidly generate mutations impacting practically any gene of interest. Unfortunately, our ability to phenotype mutant larvae has not kept pace. To address this challenge, we have developed a protocol that obtains multiple phenotypic measurements from individual zebrafish larvae in an automated and parallel fashion, including morphological features (i.e., body length, eye area, and head size) and movement/behavior. By assaying wild-type zebrafish in a variety of conditions, we determined optimal parameters that avoid significant developmental defects or physical damage; these include morphological imaging of larvae at two time points [3 days post fertilization (dpf) and 5 dpf] coupled with motion tracking of behavior at 5 dpf. As a proof-of-principle, we tested our approach on two novel CRISPR-generated mutant zebrafish lines carrying predicted null-alleles of syngap1b and slc7a5, orthologs to two human genes implicated in autism-spectrum disorder, intellectual disability, and epilepsy. Using our optimized high-throughput phenotyping protocol, we recapitulated previously published results from mouse and zebrafish models of these candidate genes. In summary, we describe a rapid parallel pipeline to characterize morphological and behavioral features of individual larvae in a robust and consistent fashion, thereby improving our ability to better identify genes important in human traits and disorders.
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影响因子：
14.9
作者：
Flicek P;Amode MR;Barrell D;Beal K;Billis K;Brent S;Carvalho-Silva D;Clapham P;Coates G;Fitzgerald S;Gil L;Girón CG;Gordon L;Hourlier T;Hunt S;Johnson N;Juettemann T;Kähäri AK;Keenan S;Kulesha E;Martin FJ;Maurel T;McLaren WM;Murphy DN;Nag R;Overduin B;Pignatelli M;Pritchard B;Pritchard E;Riat HS;Ruffier M;Sheppard D;Taylor K;Thormann A;Trevanion SJ;Vullo A;Wilder SP;Wilson M;Zadissa A;Aken BL;Birney E;Cunningham F;Harrow J;Herrero J;Hubbard TJ;Kinsella R;Muffato M;Parker A;Spudich G;Yates A;Zerbino DR;Searle SM
通讯作者：
Searle SM
定义CRISPR-CAS9全基因组核酸酶活性的活性。
影响因子：
14.8
作者：
Lazzarotto CR;Nguyen NT;Tang X;Malagon-Lopez J;Guo JA;Aryee MJ;Joung JK;Tsai SQ
通讯作者：
Tsai SQ
DOI：
10.1016/j.mcn.2018.03.008
发表时间：
2018-09
期刊：
Molecular and cellular neurosciences
影响因子：
--
作者：
Kilinc M;Creson T;Rojas C;Aceti M;Ellegood J;Vaissiere T;Lerch JP;Rumbaugh G
通讯作者：
Rumbaugh G
DOI：
10.1073/pnas.1308335110
发表时间：
2013-08-20
影响因子：
11.1
作者：
Jao, Li-En;Wente, Susan R.;Chen, Wenbiao
通讯作者：
Chen, Wenbiao
DOI：
10.1016/j.cbpc.2020.108834
发表时间：
2020-11-01
影响因子：
3.9
作者：
Felix, Luis M.;Luzio, Ana;Valentim, Ana M.
通讯作者：
Valentim, Ana M.