Dos Santos Thiciane Patrycia Gonçalves, de Melo Madson Silveira, Schramm Heloisa, Müller Yara Maria Rauh, Jaramillo Michael L B, Nazari Evelise Maria
Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil.
Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil.
Aquat Toxicol. 2020 May;222:105468. doi: 10.1016/j.aquatox.2020.105468. Epub 2020 Mar 9.
The extracellular matrix (ECM) is a non-cellular and three-dimensional structure, constituted by a macromolecular dynamic network that involves the cells in all animal tissues, including embryonic ones. Several studies with vertebrates and cell cultures have reported deleterious effects of ultraviolet-B (UVB) radiation on the components associated with the ECM. However, studies focusing on the UVB radiation effects on ECM components of crustaceans during embryonic development are very scarce. Thus, the aim of this study was to identify the coding sequences of components associated with the ECM and to evaluate the effect of UVB radiation on embryos of the ecologically-important decapod Macrobrachium olfersii. To evaluate the modulation of these ECM components during embryonic development, the transcript levels of Col4α1, Itgβ, Lamα, Mmp1 and Timp in M. olfersii embryos were analyzed at early developmental stages (E1, E3 and E4), intermediate developmental stage (E7) and late developmental stages (E10 and E14). In addition, embryos at E7, which correspond to a landmark of crustacean development, were analyzed after 12 h of UVB exposure to verify UVB effects on the ECM components. The ECM component sequences were similar to other decapods, suggesting conservation of these genes among crustaceans. The results showed modulations of the ECM components of M. olfersii embryos that reflect the need for each component in the cellular mechanisms, necessary for normal embryonic development. After UVB exposure, embryos showed opacity of embryonic tissues and it was found the overexpression of Col4α1, Itgβ, Mmp1 and Timp transcript levels (1.82-, 1.52-, 2.34- and 6.27-fold, respectively). These impairments can compromise important events for normal embryonic development, such as growth of optic lobes, caudal papilla, ramification of appendages and differentiation of organic systems. The results presented here, together with the effects on morphology, cell proliferation, differentiation, and apoptosis demonstrated previously, strengthen the knowledge of the complex impacts of UVB radiation on freshwater embryos. Nevertheless, our results encourage further investigations focusing on the assessment of UVB effects on different organisms in order to better understand the myriad of UVB effects on ECM components.
细胞外基质（ECM）是一种非细胞的三维结构，由一个大分子动态网络构成，该网络涉及所有动物组织（包括胚胎组织）中的细胞。多项针对脊椎动物和细胞培养的研究报告了紫外线B（UVB）辐射对与ECM相关成分的有害影响。然而，聚焦于UVB辐射对甲壳类动物胚胎发育过程中ECM成分影响的研究非常稀少。因此，本研究的目的是鉴定与ECM相关成分的编码序列，并评估UVB辐射对具有生态重要性的十足目动物罗氏沼虾胚胎的影响。为了评估这些ECM成分在胚胎发育过程中的调控情况，在罗氏沼虾胚胎的早期发育阶段（E1、E3和E4）、中期发育阶段（E7）以及晚期发育阶段（E10和E14）分析了Col4α1、Itgβ、Lamα、Mmp1和Timp的转录水平。此外，对处于E7阶段（对应甲壳类动物发育的一个标志性阶段）的胚胎进行UVB照射12小时后进行分析，以验证UVB对ECM成分的影响。ECM成分序列与其他十足目动物相似，表明这些基因在甲壳类动物中具有保守性。结果显示罗氏沼虾胚胎的ECM成分发生了调控，这反映了正常胚胎发育所需的细胞机制中每种成分的必要性。UVB照射后，胚胎出现胚胎组织不透明的情况，并且发现Col4α1、Itgβ、Mmp1和Timp转录水平过度表达（分别为1.82倍、1.52倍、2.34倍和6.27倍）。这些损伤可能会危及正常胚胎发育的重要事件，如视叶生长、尾乳头形成、附肢分支以及有机系统分化。此处呈现的结果，连同先前证明的对形态、细胞增殖、分化和凋亡的影响，强化了关于UVB辐射对淡水胚胎复杂影响的认识。尽管如此，我们的结果鼓励进一步开展研究，聚焦于评估UVB对不同生物体的影响，以便更好地理解UVB对ECM成分的众多影响。
