Department of Environmental Toxicology, University of California, Davis, California, United States of America.
Department of Animal Science, University of California, Davis, California, United States of America.
PLoS One. 2022 Mar 16;17(3):e0265218. doi: 10.1371/journal.pone.0265218. eCollection 2022.
Using a system optimized for propagating human keratinocytes, culture of skin samples from white and green sturgeons generated epithelial cells capable of making cross-linked protein envelopes. Two distinct forms of TGM1-like mRNA were molecularly cloned from the cells of white sturgeon and detected in green sturgeon cells, accounting for their cellular envelope forming ability. The protein translated from each displayed a cluster of cysteine residues resembling the membrane anchorage region expressed in epidermal cells of teleosts and tetrapods. One of the two mRNA forms (called A) was present at considerably higher levels than the other (called B) in both species. Continuous lines of white sturgeon epidermal cells were established and characterized. Size measurements indicated that a substantial fraction of the cells became enlarged, appearing similar to squames in human epidermal keratinocyte cultures. The cultures also expressed CYP1A, a cytochrome P450 enzyme inducible by activation of aryl hydrocarbon receptor 2 in fish. The cells gradually improved in growth rate over a dozen passages while retaining envelope forming ability, TGM1 expression and CYP1A inducibility. These cell lines are thus potential models for studying evolution of fish epidermis leading to terrestrial adaptation and for testing sturgeon sensitivity to environmental stresses such as pollution.
利用优化的人类角质形成细胞培养系统,从白鲟和绿鲟的皮肤样本中培养出能够生成交联蛋白包膜的上皮细胞。从白鲟细胞中分子克隆出两种不同形式的 TGM1 样 mRNA,并在绿鲟细胞中检测到,这解释了它们形成细胞包膜的能力。从每个细胞翻译出的蛋白质都显示出一簇半胱氨酸残基,类似于硬骨鱼和四足动物表皮细胞中表达的膜锚定区。这两种 mRNA 形式之一(称为 A)在两种物种中的表达水平明显高于另一种(称为 B)。建立并表征了连续的白鲟表皮细胞系。大小测量表明,相当一部分细胞增大,类似于人类表皮角质形成细胞培养物中的鳞屑。这些培养物还表达 CYP1A,这是一种细胞色素 P450 酶,可被鱼类芳烃受体 2 的激活所诱导。这些细胞在十多次传代过程中逐渐提高了生长速度,同时保持了形成包膜的能力、TGM1 的表达和 CYP1A 的诱导能力。因此,这些细胞系是研究鱼类表皮进化导致陆地适应的潜在模型,也是测试鲟鱼对环境污染等环境压力敏感性的模型。