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Adap1基因缺失斑马鱼的建立与鉴定

Establishment and characterization of adap1-deficient zebrafish.

作者信息

Kawahara Atsuo, Yasojima Sakyo, Koiwa Junko, Fujimaki Saori, Ito Hiroaki, Yamada Mamiko, Kosaki Kenjiro, Nishimura Yuhei

机构信息

Laboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Science, University of Yamanashi, Yamanashi, Japan.

Department of Integrative Pharmacology, Mie University Graduate School of Medicine, Tsu, Japan.

出版信息

Dev Growth Differ. 2025 Apr;67(3):165-173. doi: 10.1111/dgd.70004. Epub 2025 Mar 15.

Abstract

The adap1 (ADP-ribosylation factor GTPase-activating protein [ArfGAP] with dual pleckstrin homology [PH] domains 1) gene is predominantly expressed in the mouse brain and is important in neural differentiation and development. However, the functions of adap1 in morphogenesis, locomotor activity, and behaviors in vertebrates are not fully understood. Whole-mount in situ hybridization (WISH) analysis revealed that adap1 was widely expressed in the zebrafish brain, including the forebrain, midbrain, and hindbrain, during early embryogenesis. To investigate the physiological function of the adap1 gene, we generated zebrafish adap1 mutants harboring frameshift mutations around codon 120 of adap1. The adap1 mutants containing homozygous mutant alleles exhibited no apparent morphological abnormalities at 1 day postfertilization (dpf), and the spontaneous coiling and touch response of the adap1 mutants were comparable to those of the wild-type fish. In addition, the expression of neural genes, such as emx1, mbx, and huC, was comparable between the wild-type fish and the adap1 mutants at 1 dpf. The adap1 mutants grew to adulthood without exhibiting any apparent swimming defects. The adult adap1 mutants spent more time than the wild type in the center region of the open field test. In the social behavior test, zebrafish containing the adap1 mutant alleles spent more time than the wild type in the regions near the chambers where novel conspecifics swam. These results imply the involvement of the adap1 gene in regulating approach behavior to visual cues from conspecifics.

摘要

adap1(具有双pleckstrin同源[PH]结构域1的ADP核糖基化因子GTP酶激活蛋白[ArfGAP])基因主要在小鼠大脑中表达,对神经分化和发育很重要。然而,adap1在脊椎动物形态发生、运动活性和行为中的功能尚未完全了解。整体原位杂交(WISH)分析显示,在早期胚胎发育过程中,adap1在斑马鱼大脑中广泛表达,包括前脑、中脑和后脑。为了研究adap1基因的生理功能,我们构建了在adap1第120位密码子附近携带移码突变的斑马鱼adap1突变体。含有纯合突变等位基因的adap1突变体在受精后1天(dpf)没有明显的形态异常,adap1突变体的自发卷曲和触觉反应与野生型鱼相当。此外,在1 dpf时,野生型鱼和adap1突变体之间神经基因如emx1、mbx和huC的表达相当。adap1突变体成长至成年,未表现出任何明显的游泳缺陷。在旷场试验中,成年adap1突变体比野生型在中央区域花费更多时间。在社交行为试验中,含有adap1突变等位基因的斑马鱼比野生型在新同种个体游动的隔室附近区域花费更多时间。这些结果表明adap1基因参与调节对同种个体视觉线索的趋近行为。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/22b1/11997736/0da3bb348e0c/DGD-67-165-g004.jpg

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