Abstract
Purpose
Cadmium (Cd) causes chick embryo malformation and abnormal extra-embryonic vasculature. This study investigates the effect of Cd on vasculogenesis, quantifies extra-embryonic vascular development following exposure to cadmium acetate (CdAc).
Methods
After 48 or 60 h incubation, chicks were explanted and treated with 50 µl of 50 µM CdAc or equimolar sodium acetate. Embryos were again incubated then re-examined 4, 8, 24 and 48 h later. Gross morphological and histological manifestations were noted. Vasculogenesis was assessed by the development of omphalomesenteric vessels from blood islands. Sinus terminalis (ST), area vasculosa (AV), vessel density and embryo crown-rump length (CRL) were measured. Ang-2 and VE-cadherin mRNA expression was analysed by RT-PCR.
Results
Vasculogenesis was delayed on gross and histological examination. ST length, AV area, vessel density and CRL were significantly reduced in the Cd group. Ang-2 was increased 4 h after exposure to Cd, whereas VE-cadherin was reduced.
Conclusion
Cd exposure inhibits normal development of extra-embryonic vasculature in line with growth retardation of the chick embryo in association with altered expression of Ang-2 and VE-cadherin.
Similar content being viewed by others
Abbreviations
- AV:
-
Area vasculosa
- EEM:
-
Extra-embryonic membrane
- ST:
-
Sinus terminalis
- Cd:
-
Cadmium
- VV:
-
Vitelline vein
- RT-PCR:
-
Reverse transcription polymerase chain reaction
References
Allen WE, Wilson DJ (1993) Early embryonic angiogenesis in the chick area vasculosa. J Anat 183:579–585
Agency for Toxic Substances and Disease Registry (2008) Draft toxicological profile for cadmium. US Department of Health and Human Services, Atlanta
Bailey FR, Miller AM (1921) Textbook of embryology. Wood and Company, New York
Bridges CC, Zalups RK (2005) Molecular and ionic mimicry and the transport of toxic metals. Toxicol Appl Pharmacol 204:274–308
Carmeliet P, Lampugnani MG, Moons L, Breviario F, Compernolle V, Bono F, Balconi G, Spagnuolo F, Oosthuyse B, Dewerchin M, Zanetti A, Angellilo A, Mattot V, Nuyens D, Lutgens E, Clotman F, Dejana E (1999) Targeted deficiency or cytosolic truncation of the VE-cadherin gene in mice impairs VEGF-mediated endothelial survival and angiogenesis. Cell 98:147–157
Cheng SH, Chan PK, Wu RS (2001) The use of microangiography in detecting aberrant vasculature in zebrafish embryos exposed to cadmium. Aquat Toxicol 52:61–71
Corada M, Liao F, Lindgren M, Lampugnani MG, Breviario F, Frank R, Muller WA, Hicklin DJ, Bohlen P, Dejana E (2001) Monoclonal antibodies directed to different regions of vascular endothelial cadherin extracellular domain affect adhesion and clustering of the protein and modulate endothelial permeability. Blood 97:1679–1684
Corada M, Mariotti M, Thurston G, Smith K, Kunkel R, Brockhaus M, Lampugnani MG, Martin-Padura I, Stoppacciaro A, Ruco L, McDonald DM, Ward PA, Dejana E (1999) Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo. Proc Natl Acad Sci USA 96:9815–9820
Cullinane J, Bannigan JG, Thompson JM (2009) Cadmium teratogenesis in the chick: period of vulnerability using the early chick culture method, and prevention by divalent cations. Reprod Toxicol 28:335–341
Dejana E, Orsenigo F, Lampugnani MG (2008) The role of adherens junctions and VE-cadherin in the control of vascular permeability. J Cell Sci 121:2115–2122
Doi T, Puri P, Bannigan J, Thompson JM (2012) EphB2/B3 gene expression is down-regulated during early embryogenesis in the cadmium0induced omphalocele chick model. J Pediatr Surg 47:920–924
Doi T, Puri P, Mccann A, Bannigan J, Thompson JM (2011) Epigenetic effect of cadmium on global de novo DNA hypomethylation in the cadmium-induced ventral body wall defect (VBWD) in the chick model. Toxicol Sci 120:475–480
Drake C (2003) Embryonic and adult vasculogenesis. Birth Defects Res Part C 69:73–82
Drake CJ, Fleming PA (2000) Vasculogenesis in the day 6.5–9.5 mouse embryo. Blood 95:1671–1679
Dugan JD, Lawton MT, Glaser B, Brem H (1991) A new technique for explantation and in vitro cultivation of chicken embryos. Anat Rec 229:125–128
Eklund L, Saharinen P (2013) Angiopoietin signaling in the vasculature. Exp Cell Res 319:1271–1280
Fagiani E, Christofori G (2013) Angiopoietins in angiogenesis. Cancer Lett 328:18–26
Fernandes RA, Costola-Souza C, Sarmento COP, Goncalves L, Favaron PO, Miglino MA (2012) Placental tissues as sources of stem cells. Open J Anim Sci 2:166–173
Folkman J (1974) Tumor angiogenesis. Adv Cancer Res 19:331–358
Friberg L, Piscator M, Nordberg GF, Kjellstrom T (1974) Cadmium in the environment. Cleveland, OH, CRC pess, p 94
Fujii T, Kuwano H (2010) Regulation of the expression balance of angiopoietin-1 and angiopoietin-2 by Shh and FGF-2. In vitro cellular and developmental biology. Animal 46:487–491
Gheorghescu A, Tywoniuk B, Duess J, Buchete NV, Thompson J (2015) Exposure of chick embryos to cadmiuml changes the extra-embryonic vascular branching pattern and alters expression of VEGF-A and VEGF-R2. Toxicol Appl Pharmacol 269:79–88
Giles JJ, Bannigan JG (1999) The effects of lithium on vascular development in the chick area vasculosa. J Anat 194:197–205
Gonzalez-Crussi F (1971) Vasculogenesis in the chick embryo. An ultrastructural study. Am J Anat 130:441–460
Greene AS (1998) Microvascular remodeling in hypertension: a role for the renin-angiotensin aldosterone system. Curr Concepts Hypertens 2:5
Halder A (2010) Amniotic band syndrome and/or limb body wall complex: split or lump. Clin Appl Clin Genet 3:7–15
Hamburger H, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Hanahan D (1997) Signaling vascular morphogenesis and maintenance. Science 277:48–50
Hoey MJ (1966) The effects of metallic salts on the histology and functioning of the rat testis. J Reprod Fertil 12:461–472
Hoper J, Jahn H (1995) Influence of environmental oxygen concentration on growth and vascular density of the area vasculosa in chick embryos. Int J Microcirc 15:186–192
Hoyme HE, Higginbottom MC, Jones KL (1981) The vascular pathogenesis of gastroschisis: intrauterine interruption of the omphalomesenteric artery. J Pediatr Surg 98:228–231
IARC (1993) Beryllium, cadmium, mercury and exposure in the glass industry. IARC, Lyon
IARC (1993b) Monographs on the evaluation of carcinogenic risks to humans
Ikeh-Tawari EP, Anetor JI, Charles-Davies MA (2013) Cadmium level in pregnancy, influence on neonatal birth weight and possible amelioration by some essential trace elements. Toxicol Int 20:108–112
Jing Y, Liu L, Jiang Y, Zhu Y, Guo NL, Barnett JB, Rojanasakul Y, Agani F, Jiang B (2012) Cadmium increases HIF-1 and VEGF expression through ROS, ERK, and AKT signaling pathways and induces malignant tranformation of human bronchial epithelial cells. Toxicol Sci 125:10–19
Kar AB, Das RP (1960) Testicular changes in rats after treatment with cadmium chloride. Acta Biol Medica Ger 5:153–173
Kim J, Lim W, Ko Y, Kwon H, Kim S, Kim O, Parl G, Choi H, Kim O (2012) The effects of cadmium on VEGF-mediated angiogenesis in HUVECs. J Appl Toxicol 32:342–349
Kirchner LM, Schmidt P, Gruber BS (1996) Quantification of angiogenesis in the chick chorioallantoic membrane model using fractal analysis. Microvasc Res 51:2–14
Kishimoto T, Fukuzawa Y, Abe M, Isobe M, Hashimoto M, Tada M (1991) Cadmium injury of cultured human vascular endothelial cells. Hum Cells 4:329–334
Kishimoto T, Oguri T, Ohno M, Matsubara K, Yamamoto K, Tada M (1994) Effect of cadmium (CdCl2) on cell proliferation and production of EDRF (endothelium-derived relaxing factor) by cultured human arterial andothelial cells. Arch Toxicol 68:555–559
Kishimoto T, Ueda D, Isobe M, Tada M (1996) Cadmium injuries tube formation by cultured human vascular endothelial cells. Hum Cell 9:244–250
Lubinsky M (2014) A vascular and thrombotic model of gastroschisis. Am J Med Genet 164A:915–917
Mannino DM, Holguin F, Greves HM, Savage-Brown A, Stock AL, Jones RL (2004) Urinary cadmium levels predict lower lung function in current and former smokers: data from the Third National Health and Nutrition Examination Survey. Thorax 59:194–198
Marshak DR, Gardner RL, Gottlieb D (2001) Stem cell biology. Cold Spring Harbor Laboratory Press
Menai M, Heude B, Slama R, Forhan A, Sahuquillo J, Charles-Davies MA, Yazbeck C (2012) Association between maternal blood cadmium during pregnancy and birth weight and the risk of fetal growth restriction: the EDEN mother-child cohort study. Reprod Toxicol 34:622–627
Messerle K, Webster WS (1982) The classification and dedevelopment of cadmium-induced limb defects in mice. Teratology 25:61–70
Nasreddine L, Parent-Massin D (2002) Food contamination by metals and pesticides in the European Union. Should we worry? Toxicol Lett 127:29–41
Niewenhuis RJ, Dimitriu C, Prozialeck WC (1997) Ultrastructural characterization of the early changes in intercellular junctions in response to cadmium (Cd2+) exposure in LLC-PK1 cells. Toxicol Appl Pharmacol 142:1–12
Nilsen NO (1984) Vascular abnormalities due to hyperthermia in chick embryos. Teratology 30:237–251
Parsons-Wingerter P, Lwai B, Yang MC, Elliot KE, Milaninia A, Redlitz A, Clark JI, Sage EH (1998) A novel assay of angiogenesis in the quail chorioallantoic membrane: stimulation of bFGF and inhibition by angiostatin according to fractal dimension and grid intersection. Microvasc Res 55:201–214
Partanen J, Armstrong E, Makela TP, Korhonen J, Sandberg M, Renkonen R, Knuutila S, Huebner K, Alitalo K (1992) A novel endothelial cell surface receptor tyrosine kinase with extracellular epidermal growth factor homology domains. Mol Cell Biol 12:1698–1707
Pearson CA, Lamar PC, Prozialeck WC (2003) Effects of cadmium on E-cadherin and VE-cadherin in mouse lung. Life Sci 72:1303–1320
Peault B (2010) Haemangioblasts: back to the future? Blood 116:2864–2966
Peifer C, Dannhardt G (2004) A novel quantitative chick embryo assay as an angiogenesis model using digital image analysis. Anticancer Res 24:1545–1552
Prozialeck WC (2000) Evidence that E-cadherin may be a target for cadmium toxicity in epithelial cells. Toxicol Appl Pharmacol 164:231–249
Prozialeck WC, Edwards JR, Nebert DW, Woods JM, Barchowsky A, Atchison WD (2008) The vascular systems as a target of metal toxicity. Toxicol Sci 102:207–218
Prozialeck WC, Edwards JR, Woods JM (2006) The vascular endothelium as a target of cadmium toxicity. Life Sci 79:1493–1506
Prozialeck WC, Niewenhuis RJ (1991) Cadmium (Cd2+) disrupts Ca(2+)-dependent cell-cell junctions and alters the pattern of E-cadherin immunofluorescence in LLC-PK1 cells. Biochem Biophys Res Commun 181:1118–1124
Ramezani M, Bahadoran H, Abasi S (2011) Effect of L-carnitin on cadmium induced toxicity in rat embryo hippocampus. Int Proc Chem Biol Environ Eng 19:147–150
Ribatti D, Vacca A, Roncali L, Dammacco F (1996) The chick embryo chorioallantoic membrane as a model for in vivo research on angiogenesis. Int J Biol 40:1189–1197
Rieder MJ, O’drobinak DM, Greene AS (1995) A computerized method for determination of microvascular density. Microvasc Res 49:180–189
Risau W (1991) Vasculogenesis, angiogenesis and endothelial cell differentiation during embryonic development. Issues Biomed 14:58–68
Risau W (1997) Mechanisms of angiogenesis. Nature 386:671–674
Risau W, Flamme I (1995) Vasculogenesis. Annu Rev Cell Dev Biol 11:73–91
Samarawickrama GP, Webb M (1981) The acute toxicity and teratogenicity of cadmium in the pregnant rat. J Appl Toxicol 1:264–269
Sato TN, Tozawa Y, Deutsh U, Wolburg-Buchholz K, Fujiwara M, Gendron-Maguire T, Gridley T, Wolburg H, Risau W, Qin Y (1995) Distinct roles of the receptor tyrosine kinases Tie-1 and Tie-2 in blood vessel formation. Nature 376:70–74
Schmidt A, Brixius K, Bloch W (2007) Endothelial precursor cell migration during vasculogenesis. Circ Res 101:125–136
Sorokin VA, Valeev VA, Gladchenko GO, Sysa IV (1997) Interaction of divalent cadmium ions with nucleotides and native DNA. Biofizika 42:105–116
Thompson JM, Bannigan JG (2001) The effects of cadmium on formation of the ventral body wall in chick embryos and their prevention by zinc pre-treatment. Teratology 64:87–97
Thompson JM, Bannigan JG (2007) Omphalocele induction in the chick embryo by administration of cadmium. J Pediatr Surg 42:1703–1709
Thompson JM, Doi T, Power E, Balasubramanian I, Puri P, Bannigan J (2010) Evidence against a direct role for oxidative stress in cadmium-induced axial malformation in the chick embryo. Toxicol Appl Pharmacol 243:390–398
Valois AA, Webster WS (1987) Retention and distribution of cadmium in the mouse brain: an autoradiographic and gamma counting study. Neurotoxicology 8:463–470
Valois AA, Webster WS (1989) The choroid plexus as a target site for cadmium toxicity following chronic exposure in the adult mice: an ultrastructural study. Toxicology 55:193–205
Vaois AA, Webster WS (1987) The choroid plexus and cerebral vasculature as target sites for cadmium following acute exposure in neonatal and adult mice: an autoradiographic and gamma counting study. Toxicology 46:43–55
Velde EA, Exalto N, Hesseling P, Linden HC (1997) First trimester development of human chorionic villous vascularization studied with CD34 immunohistochemistry. Hum Reprod 12:1577–1581
Vico PG, Kyriacos SOH, Louryan S, Cartilier LH (1998) Dynamic study of the extra-embryonic vascular network of the chick embryo by fractal analysis. J Theor Biol 95:525–532
Webster WAS, Messerle K (1980) Changes in the mouse neuroepithelium associated with cadmium-induced neural tube defects. Teratology 21:79–88
Webster WS (1988) Chronic cadmium exposure during pregnancy in the mouse: influence of exposure levels on fetal and maternal uptake. J Toxicol Environ Health 24:183–192
Werler MM, Mitchell AA, Moore CA, Honein MA (2009) It there epidemiologic evidence to support vascular disruption as a pathogenesis of gastroschisis? Am J Med Genet 149A:1399–1406
WHO (1993) Evaluation of certain food additives and contaminants (Fourty-first Report of the Joint FAO/WHO Expert Committee on Food Additives). WHO Technical Report Series No. 837
Woods JM, Leone M, Klosowska K, Lamar PC, Shaknovsky TJ, Prozialeck WC (2008) Direct antiangiogenic actions of cadmium on human vascular endothelial cells. Toxicol Vitro 22:643–651
Yamamoto FY, Neto FF, Freitas PF, Ribeiro CAO (2012) Cadmium effects on early development of chick embryos. Environ Toxicol Pharmacol 34:548–555
Yuan HT, Khankin EV, Karumanchi SA, Parikh SM (2009) Angiopoietin 2 is a partial agonist/antagonist of Tie2 signaling in the endothelium. Mol Cell Biol 29:2011–2022
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing financial interest.
Rights and permissions
About this article
Cite this article
Gheorghescu, A., Thompson, J. Delayed vasculogenesis and impaired angiogenesis due to altered Ang-2 and VE-cadherin levels in the chick embryo model following exposure to cadmium. Pediatr Surg Int 32, 175–186 (2016). https://doi.org/10.1007/s00383-015-3830-9
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00383-015-3830-9