TY - JOUR
T1 - Embryotoxicity of silver nanomaterials (Ag NM300k) in the soil invertebrate Enchytraeus crypticus – Functional assay detects Ca channels shutdown
AU - Bicho, Rita C.
AU - Faustino, Augusto M.R.
AU - Carvalho, Fátima
AU - Soares, Amadeu M.V.M.
AU - Scott-Fordsmand, Janeck J.
AU - Amorim, Mónica J.B.
N1 - Funding Information:
This study was supported by the European Commission Projects: BIORIMA - BIOmaterial RIsk MAnagement (H2020-NMBP2017, GA No. 760928), NanoInformaTIX - Development and Implementation of a Sustainable Modelling Platform for NanoInformatics (H2020-NMBP-14-2018, GA No. 814426) and NANORIGO - Establishing a Nanotechnology Risk Governance Framework (H2020-NMBP-TO-IND-2018, GA No. 814530). Further support within CESAM (UIDB/50017/2020+UIDP/50017/2020), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020, and by FCT (Funda??o para a Ci?ncia e Tecnologia) via the individual PhD grant to Rita Bicho (SFRH/BD/102702/2014).
Funding Information:
This study was supported by the European Commission Projects: BIORIMA - BIOmaterial RIsk MAnagement ( H2020-NMBP2017 , GA No. 760928 ), NanoInformaTIX - Development and Implementation of a Sustainable Modelling Platform for NanoInformatics ( H2020-NMBP-14-2018 , GA No. 814426 ) and NANORIGO - Establishing a Nanotechnology Risk Governance Framework ( H2020-NMBP-TO-IND-2018 , GA No. 814530 ). Further support within CESAM ( UIDB/50017/2020+UIDP/50017/2020 ), to FCT/MEC through national funds, and the co-funding by the FEDER , within the PT2020 Partnership Agreement and Compete 2020, and by FCT (Fundação para a Ciência e Tecnologia) via the individual PhD grant to Rita Bicho ( SFRH/BD/102702/2014 ).
Publisher Copyright:
© 2021 Elsevier B.V.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/1
Y1 - 2021/1
N2 - Despite that silver (Ag) is among the most studied nanomaterials (NM) in environmental species and Ag's embryotoxicity is well known, there are no studies on Ag NMs embryotoxicity in soil invertebrates. Previous Full Life Cycle (FLC) studies in Enchytraeus crypticus, a standard soil invertebrate, showed that Ag materials decreased hatching success, which was confirmed to be a hatching delay effect for silver nitrate (AgNO3) and mortality for Ag NM300K. In the present study, we aimed to investigate if the impact of Ag takes place during the embryonic development, using histology and immunohistochemistry. E. crypticus cocoons were exposed to a range of concentrations of Ag NM300K (0–10–20–60–115 mg Ag/kg) and AgNO3 (0–20–45–60–96 mg Ag/kg) in LUFA 2.2 soil, in an embryotoxicity test, being sampled at days 1, 2, 3 and 6 (3, 4, 5 and 7 days after cocoon laying). Measured endpoints included the number of embryonic structures, expression of transferrin receptor (TfR) and L type calcium channels (LTCC) through histological and immunohistochemistry analysis, respectively. Results confirmed that Ag materials affected the embryonic development, specifically at the blastula stage (day 3). The expression and localization of TfR in E. crypticus was shown in the teloblasts cells, although this transcytosis mechanism was not activated. Ag affected calcium (Ca) metabolism during embryonic development: for AgNO3, LTCC was initially activated, compensating the impact, for Ag NM300K, LTCC was not activated, hence no Ca balance, with irreversible consequences, i.e. terminated embryonic development. An Adverse Outcome Pathway was drafted, integrating the mechanisms here discovered with previous knowledge.
AB - Despite that silver (Ag) is among the most studied nanomaterials (NM) in environmental species and Ag's embryotoxicity is well known, there are no studies on Ag NMs embryotoxicity in soil invertebrates. Previous Full Life Cycle (FLC) studies in Enchytraeus crypticus, a standard soil invertebrate, showed that Ag materials decreased hatching success, which was confirmed to be a hatching delay effect for silver nitrate (AgNO3) and mortality for Ag NM300K. In the present study, we aimed to investigate if the impact of Ag takes place during the embryonic development, using histology and immunohistochemistry. E. crypticus cocoons were exposed to a range of concentrations of Ag NM300K (0–10–20–60–115 mg Ag/kg) and AgNO3 (0–20–45–60–96 mg Ag/kg) in LUFA 2.2 soil, in an embryotoxicity test, being sampled at days 1, 2, 3 and 6 (3, 4, 5 and 7 days after cocoon laying). Measured endpoints included the number of embryonic structures, expression of transferrin receptor (TfR) and L type calcium channels (LTCC) through histological and immunohistochemistry analysis, respectively. Results confirmed that Ag materials affected the embryonic development, specifically at the blastula stage (day 3). The expression and localization of TfR in E. crypticus was shown in the teloblasts cells, although this transcytosis mechanism was not activated. Ag affected calcium (Ca) metabolism during embryonic development: for AgNO3, LTCC was initially activated, compensating the impact, for Ag NM300K, LTCC was not activated, hence no Ca balance, with irreversible consequences, i.e. terminated embryonic development. An Adverse Outcome Pathway was drafted, integrating the mechanisms here discovered with previous knowledge.
KW - Adverse Outcome Pathway (AOP)
KW - Calcium metabolism
KW - Embryonic development
KW - Oligochaete
KW - Transcytosis
UR - https://www.scopus.com/pages/publications/85103114591
U2 - 10.1016/j.impact.2021.100300
DO - 10.1016/j.impact.2021.100300
M3 - Journal article
C2 - 35559787
AN - SCOPUS:85103114591
SN - 2452-0748
VL - 21
JO - NanoImpact
JF - NanoImpact
M1 - 100300
ER -