Finally, the thyroid dysplasia in isl2a mutant larvae may be attributed to a reduction in proliferation rather than changes in apoptosis.Ĭonclusions: In summary, isl2a regulates the transcriptional levels of marker genes in hypothalamus-pituitary-thyroid axis, and isl2a knockout causing low thyroid hormone levels in zebrafish. Importantly, the transcriptional levels of pituitary-thyroid axis hormones-encoding genes, such as tshba, cga, and tg, were significantly decreased in isl2a mutants. Additionally, thyrotropes, a pituitary cell type, was notably decreased during development. Results: The isl2a mutant zebrafish presented with thyroid hypoplasia, reduced whole-body levels of thyroid hormones, increased early mortality, gender imbalance, and morphological retardation during maturity. For further molecular characterization, in situ hybridization and immunofluorescence were performed. Moreover, we utilized these knockout zebrafish to analyze the pituitary and thyroid phenotypes in vivo. Methods: We used the CRISPR/Cas9 system to successfully establish homozygous ISL2-orthologue ( isl2a and isl2b) knockout zebrafish. In the present study, we characterized the expression patterns of ISL2 and revealed its regulative role during embryogenesis using zebrafish. However, the role of ISL2 in the hypothalamus-pituitary-thyroid axis is largely unknown. 2Geriatric Medicine Center, Department of Endocrinology, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, Chinaīackground: ISL LIM homeobox 2, also known as insulin gene enhancer protein ISL-2 ( ISL2), is a transcription factor gene that participates in a wide range of developmental events.1Department of Molecular Diagnostics and Endocrinology, The Core Laboratory in Medical Center of Clinical Research, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University (SJTU) School of Medicine, Shanghai, China.Chen-Yan Yan 1,2†, Feng-Yao Wu 1†, Feng Sun 1, Ya Fang 1, Rui-Jia Zhang 1, Chang-Run Zhang 1, Cao-Xu Zhang 1, Zheng Wang 1, Rui-Meng Yang 1, Liu Yang 1, Mei Dong 1, Qian-Yue Zhang 1, Xiao-Ping Ye 1, Huai-Dong Song 1* and Shuang-Xia Zhao 1*
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