Department of Molecular and Translational Medicine, Division of Biology and Genetics, University of Brescia, 25123 Brescia, Italy.
Department of Molecular and Translational Medicine, Division of Experimental Oncology and Immunology, University of Brescia, 25123 Brescia, Italy.
Genes (Basel). 2022 Dec 22;14(1):35. doi: 10.3390/genes14010035.
The LIM and SH3 domain protein 1 (LASP1) was originally identified in metastatic breast cancer and mainly characterized as a cytoskeleton protein overexpressed in various cancer types. At present, little is known about LASP1 expression in physiological conditions, and its function during embryonic development has not been elucidated. Here, we focused on Lasp1 and embryonic development, choosing zebrafish as a vertebrate model. For the first time, we identified and determined the expression of Lasp1 protein at various stages of development, at 48 and 72 h post-fertilization (hpf), at 6 days pf and in different organs of zebrafish adults by Western blotting, 3D light-sheet microscopy and fluorescent immunohistochemistry. Further, we showed that specific morpholino (MO) led to (i) abnormal morphants with alterations in several organs, (ii) effective knockdown of endogenous Lasp1 protein and (iii) an increase in mRNA, as detected by ddPCR. The co-injection of mRNA with MO partially rescued morphant phenotypes, thus confirming the specificity of the MO oligonucleotide-induced defects. We also detected an increase in apoptosis following MO treatment. Our results suggest a significant role for Lasp1 in embryonic development, highlighting zebrafish as a vertebrate model suitable for studying Lasp1 function in developmental biology and organogenesis.
LIM 和 SH3 域蛋白 1（LASP1）最初在转移性乳腺癌中被发现，主要特征是在各种癌症类型中过度表达的细胞骨架蛋白。目前，人们对 LASP1 在生理条件下的表达知之甚少，其在胚胎发育过程中的功能也尚未阐明。在这里，我们专注于 Lasp1 和胚胎发育，选择斑马鱼作为脊椎动物模型。我们首次通过 Western blot、3D 光片显微镜和荧光免疫组织化学技术，在受精后 48 和 72 小时（hpf）、6 天时以及成年斑马鱼的不同器官中鉴定并确定了 Lasp1 蛋白的表达。此外，我们还表明，特异性的 morpholino（MO）导致（i）几种器官发生改变的异常畸形，（ii）内源性 Lasp1 蛋白的有效敲低，以及（iii）通过 ddPCR 检测到的 mRNA 增加。用 mRNA 与 MO 共注射部分挽救了畸形表型，从而证实了 MO 寡核苷酸诱导缺陷的特异性。我们还检测到 MO 处理后细胞凋亡增加。我们的研究结果表明，Lasp1 在胚胎发育中起着重要作用，突显了斑马鱼作为一种适合研究 Lasp1 在发育生物学和器官发生中的功能的脊椎动物模型。
