Vitorino H A, Alta R Y P, Alta E C P, Ortega P
Center of Natural and Human Sciences, Federal University of ABC, Santo André, Brazil.
Faculty of Science, National University of Engineering, Lima, Peru.
In Vitro Cell Dev Biol Anim. 2017 Oct;53(9):778-781. doi: 10.1007/s11626-017-0168-8. Epub 2017 Jun 7.
Crustaceans are frequently used as bioindicators, and changes in their metabolism at the hepatopancreas (HP) level are often followed in these studies. The HP is the site of digestion, absorption, nutrient storage, and toxic metal detoxification, enabling crabs to survive in metal contaminated regions. Cellular damage and high lipid peroxidation (LPO) levels have been found in crab populations under high cadmium (Cd) concentrations. The aim of this work was to separate and characterize the HP cells of the stone crab Menippe frontalis from the Pacific Ocean, Perú (5° 5' 21″ S-81° 6' 51″ W) and to measure the cellular viability and LPO after exposure to the non-essential metal Cd. The HP cells were dissociated by magnetic stirring, with posterior separation by sucrose gradient at concentrations of 10, 20, 30, and 40%. We found the same cell types that were described for other species (e.g., Ucides cordatus, Atlantic Ocean, Brazil). High cellular viability against 1 mmol L of Cd was observed for resorptive (R) cells in 20% sucrose layer (88 ± 8%, *P < 0.05, ANOVA), and blister (B) cells in the 40% sucrose layers (92 ± 7%, *P < 0.05, ANOVA). Cd (1 mmol L) caused an increase in LPO levels, suggesting that crabs from polluted areas can be affected by toxic metals, generating a physiological stress. The gradient sucrose methodology can be used for different species and results in a similar separation, viability, and cellular identification. The results are a starting point for toxic metal studies for species distributed across different geographic coordinates.
甲壳类动物常被用作生物指示物,在这些研究中,它们肝胰腺(HP)水平的代谢变化经常受到关注。肝胰腺是消化、吸收、营养储存和有毒金属解毒的场所,使螃蟹能够在金属污染区域生存。在高镉(Cd)浓度下的螃蟹种群中发现了细胞损伤和高脂质过氧化(LPO)水平。这项工作的目的是分离和鉴定来自秘鲁太平洋地区(南纬5°5′21″-西经81°6′51″)的石蟹Menippe frontalis的肝胰腺细胞,并测量暴露于非必需金属Cd后的细胞活力和LPO。通过磁力搅拌解离肝胰腺细胞,然后用浓度为10%、20%、30%和40%的蔗糖梯度进行后续分离。我们发现了与其他物种(如巴西大西洋的Ucides cordatus)所描述的相同细胞类型。在20%蔗糖层中,吸收性(R)细胞对1 mmol/L的Cd具有较高的细胞活力(88±8%,*P<0.05,方差分析),在40%蔗糖层中的泡状(B)细胞也是如此(92±7%,*P<0.05,方差分析)。1 mmol/L的Cd导致LPO水平升高,表明来自污染地区的螃蟹可能受到有毒金属的影响,产生生理应激。梯度蔗糖方法可用于不同物种,并能实现类似的分离、活力和细胞鉴定。这些结果是对分布在不同地理坐标的物种进行有毒金属研究的起点。