Premium
Primary culture of isolated cardiac myocytes from the bullfrog, Rana catesbeiana
Author(s) -
Duerr Jeffrey M,
Luft Silvianne
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.778.9
Subject(s) - myocyte , bullfrog , biology , rana , contractility , microbiology and biotechnology , cardiac muscle , medicine , endocrinology
Comparative vertebrate studies into physiological responses to adverse conditions can yield great insight into the design of robust systems and provide fundamental knowledge of stress adaptations. The classically studied cellular stress response is the mitogen‐activated protein kinase (MAPK) pathway. Prior investigations in our laboratory indicated a differential response of MAPK family activation to progressive dehydration in cardiac tissue from two species of anuran amphibians: Rana catesbeiana and Bufo marinus. Western blot analysis indicated that pERK increased by three‐fold in Bufo and 2‐fold in Rana. pJNK levels remained unchanged in Bufo whereas in Rana increased by 50%. pp38 did not exhibit any change in phosphorylation levels over the range of dehydration examined. To more specifically delineate signaling pathways activated in response to dehydrational stress (hypoxia, hyperosmolality) in anuran cardiac tissue, we adapted a variety of previously described methodologies to develop a reliable technique for isolating and retaining in primary culture cardiac myocytes from Rana catesbeiana. Isolated myocytes exhibited mostly a spindle‐like morphology with characteristic banding patterns and occasional spontaneous contractility when viewed under phase‐contrast microscopy. Myocytes were also immunostained with a fluorescent monoclonal antibody against the SR ryanodine receptor.