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−1 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−1) 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.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Al-Mohanna SY, Nott JA (1989) Functional cytology of the hepatopancreas of Penaeus semisulcatus (Crustacea: Decapoda) during the moult cycle. Mar Biol 101(4):535–544
Chavez-Crooker P, Garrido N, Ahearn GA (2001) Copper transport by lobster hepatopancreatic epithelial cells separated by centrifugal elutriation: measurements with the fluorescent dye Phen green. J Exp Biol 204(8):1433–1444
Harris RR, Santos MCF (2000) Heavy metal contamination and physiological variability in the Brazilian mangrove crabs Ucides cordatus and Callinectes danae (Crustacea: Decapoda). Mar Biol 137:691–703
Hermes-Lima M, Willmore WG, Storey KB (1995) Quantification of lipid peroxidation in tissue extracts based on Fe(III)xylenol orange complex formation. Free Radic Biol Med 19(3):271–280
Hopkin SP, Nott JA (1980) Studies on the digestive cycle of the shore crab Carcinus maenas (L.) with special reference to the b cells in the hepatopancreas. J Mar Biol Assoc U K 60(4):891
Jiang Z-Y, Hunt JV, Wolff SP (1992) Ferrous ion oxidation in the presence of xylenol orange for detection of lipid hydroperoxide in low density lipoprotein. Anal Biochem 202(2):384–389
Jiang Z-Y, Woollard ACS, Wolff SP (1991) Lipid hydroperoxide measurement by oxidation of Fe2+ in the presence of xylenol orange. Comparison with the TBA assay and an iodometric method. Lipids 26(10):853–856
Lan W, Dingxing W, Qian W, Xiuqing Y (2004) Effect of cadmium on ultrastructure of hepatopancreas cells of freshwater crab (Sinopotamon yangtsekiense). Acta Anatomica Sinica 34(5):522–526
Monserrat JM, Geracitano LA, Pinho GLL, Vinagre TM, Faleiros M, Alciati JC, Bianchini A (2003) Determination of lipid peroxides in invertebrates tissues using the Fe(III) xlenol orange complex formation. Arch Environ Contam Toxicol 45(2):177–183
Ortega P, Custódio MR, Zanotto FP (2014b) Characterization of cadmium plasma membrane transport in gills of a mangrove crab Ucides cordatus. Aquat Toxicol 157:21–29
Ortega P, Granado e Sá M, Custódio MR, Zanotto FP (2011) Separation and viability of gill and hepatopancreatic cells of a mangrove crab Ucides cordatus. In Vitro Cell Dev Biol Anim 47(5–6):346–349
Ortega P, Santos RA, Lacouth P, Rozas EE, Custódio MR, Zanotto FP (2014a) Cytochemical characterization of gill and hepatopancreatic cells of the crab Ucides cordatus (Crustacea, Brachyura) validated by cell metal transport. Iheringia Série Zoologia 104(3):347–354
Ortega P, Vitorino HA, Moreira RG, Pinheiro MAA, Almeida AA, Custódio MR, Zanotto FP (2016) Physiological differences in the crab Ucides cordatus from two populations inhabiting mangroves with different levels of cadmium contamination. Environ Toxicol Chem 9999(9999):1–11
Rinderhagen M, Ritterhoff J, Zauke GP (2000) Crustaceans as Bioindicators. In: Biomonitoring of polluted water-reviews on actual topics. Trans Tech Publications-Scitech Publications, Environmental Research Forum, pp 161–194
Rotter S, Sans-Piché F, Streck G, Altenburger R, Schmitt-Jansen M (2010) Active bio-monitoring of contamination in aquatic systems—an in situ translocation experiment applying the PICT concept. Aquat Toxicol 101:228–236
Silvestre F, Trausch G, Pequeux A, Devos P (2004) Uptake of cadmium through isolated perfused gills of the Chinese mitten crab, Eriocheir sinensis. Comparative Biochemistry and Physiology Part A 137:189–196
Tennant J (1964) Evaluation of the Trypan Blue technique for determination of cell viability. Transplantation 2(6):685–694
Vitorino HA, Mantovanelli L, Zanotto FP, Espósito BP (2015) Iron metallodrugs: stability, redox activity and toxicity against Artemia salina. PLoS One 10(4):1–11
Vogt G (1994) Life-cycle and functional cytology of the hepatopancreatic cells of Astacus astacus (Crustacea, Decapoda). Zoomorphology 114(2):83–101
Wang Y, Wu Y, Luo K, Liu Y, Zhou M, Yan S, Shi H, Cai Y (2013) The protective effects of selenium on cadmium-induced oxidative stress and apoptosis via mitochondria pathway in mice kidney. Food Chem Toxicol 58:61–67
Xuan R, Wang L, Sun M, Ren G, Jiang M (2011) Effects of cadmium on carbohydrate and protein metabolisms in the freshwater crab Sinopotamon yangtsekiense. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 154(3):268–274
Yang J, Liu D, Jing W, Dahms H-U, Wang L (2013) Effects of cadmium on lipid storage and metabolism in the freshwater crab Sinopotamon henanense. PLoS One 8(10):1–10
This work was supported by funds from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and the Universidad Nacional de Ingeniería, Facultad de Ciencias (FC-UNI). The authors thank Dr. Breno Pannia Espósito, Dr. Flavia Pinheiro Zanotto and Mr. Alejandro J. Aranda Aguirre for discussions and technical assistance.
The original version of this article was revised: The correct names of the second and third authors are R. Y. P. Alta and E. C. P. Alta, respectively.
An erratum to this article is available at https://doi.org/10.1007/s11626-017-0182-x.
About this article
Cite this article
Vitorino, H.A., Alta, R.Y.P., Alta, E.C.P. et al. Hepatopancreatic cells of a stone crab Menippe frontalis from Perú: separation, viability study, and evaluation of lipoperoxidation against cadmium contamination. In Vitro Cell.Dev.Biol.-Animal 53, 778–781 (2017). https://doi.org/10.1007/s11626-017-0168-8
- Menippe Frontalis
- Isolated cells
- Lipid peroxidation