caa a22 4500 445823151 CHVBK 20180317145250.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10616-011-9347-8 doi (NATIONALLICENCE)springer-10.1007/s10616-011-9347-8 Establishment of functional primary cultures of heart cells from the clam Ruditapes decussatus [Elektronische Daten] [H. Hanana, H. Talarmin, J. Pennec, M. Droguet, E. Gobin, P. Marcorelle, G. Dorange] Heart cells from the clam Ruditapes decussatus were routinely cultured with a high level of reproducibility in sea water based medium. Three cell types attached to the plastic after 2days and could be maintained in vitro for at least 1month: epithelial-like cells, round cells and fibroblastic cells. Fibroblastic cells were identified as functional cardiomyocytes due to their spontaneous beating, their ultrastructural characteristics and their reactivity with antibodies against sarcomeric α-actinin, sarcomeric tropomyosin, myosin and troponin T-C. Patch clamp measurements allowed the identification of ionic currents characteristic of cardiomyocytes: a delayed potassium current (I Kslow) strongly suppressed (95%) by tetraethylammonium (1mM), a fast inactivating potassium current (I Kfast) inhibited (50%) by 4 amino-pyridine at 1mM and, at a lower level (34%) by TEA, a calcium dependent potassium current (I KCa) activated by strong depolarization. Three inward voltage activated currents were also characterized in some cardiomyocytes: L-type calcium current (I Ca) inhibited by verapamil at 5×10−4M, T-type Ca2+ current, rapidly activated and inactivated, and sodium current (I Na) observed in only a few cells after strong hyperpolarization. These two currents did not seem to be physiologically essential in the initiation of the beatings of cardiomyocytes. Potassium currents were partially inhibited by tributyltin (TBT) (1μM) but not by okadaic acid (two marine pollutants). DNA synthesis was also demonstrated in few cultured cells using BrdU (bromo-2′-deoxyuridine). Observed effects of okadaic acid and TBT demonstrated that cultured heart cells from clam Ruditapes decussatus can be used as an experimental model in marine toxicology. Springer Science+Business Media B.V., 2011 Clam nationallicence Ruditapes decussatus nationallicence Cardiomyocyte nationallicence Patch clamp nationallicence Ionic currents nationallicence In vitro nationallicence Hanana H. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut Talarmin H. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut Pennec J. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut Droguet M. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut Gobin E. CHU Morvan, Service d'Anatomie Pathologique, 29609, Brest, France aut Marcorelle P. CHU Morvan, Service d'Anatomie Pathologique, 29609, Brest, France aut Dorange G. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut Cytotechnology Springer Netherlands 63/3(2011-05-01), 295-305 0920-9069 63:3<295 2011 63 10616 https://doi.org/10.1007/s10616-011-9347-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9347-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hanana H. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut NATIONALLICENCE 700 1- Talarmin H. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut NATIONALLICENCE 700 1- Pennec J. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut NATIONALLICENCE 700 1- Droguet M. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut NATIONALLICENCE 700 1- Gobin E. CHU Morvan, Service d'Anatomie Pathologique, 29609, Brest, France aut NATIONALLICENCE 700 1- Marcorelle P. CHU Morvan, Service d'Anatomie Pathologique, 29609, Brest, France aut NATIONALLICENCE 700 1- Dorange G. Faculté de médecine, EA 4326, Université Européenne de Bretagne, Université de Bretagne Occidentale, 22 Avenue Camille Desmoulins, 29238, Brest Cedex 3, France aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/3(2011-05-01), 295-305 0920-9069 63:3<295 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer