Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)
Abstract Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and b...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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Online Access: | https://doi.org/10.1590/1678-9199-jvatitd-2019-0078 https://doaj.org/article/5c0e65622aa24f33b5008a09c4c24385 |
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ftdoajarticles:oai:doaj.org/article:5c0e65622aa24f33b5008a09c4c24385 2023-05-15T15:13:34+02:00 Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) Luciano S. Fusco Rodrigo Cajade Jose M. Piñeiro Ana M. Torres Igor R. F. da Silva Stephen Hyslop Laura C. Leiva Daniel C. Pimenta Soledad Bustillo 2020-03-01T00:00:00Z https://doi.org/10.1590/1678-9199-jvatitd-2019-0078 https://doaj.org/article/5c0e65622aa24f33b5008a09c4c24385 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100306&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20190078.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2019-0078 https://doaj.org/article/5c0e65622aa24f33b5008a09c4c24385 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) Argenteohyla siemersi Hylidae Skin secretion Tree frog Cytotoxicity Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2020 ftdoajarticles https://doi.org/10.1590/1678-9199-jvatitd-2019-0078 2022-12-31T16:03:45Z Abstract Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Argentina Journal of Venomous Animals and Toxins including Tropical Diseases 26 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Argenteohyla siemersi Hylidae Skin secretion Tree frog Cytotoxicity Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
Argenteohyla siemersi Hylidae Skin secretion Tree frog Cytotoxicity Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Luciano S. Fusco Rodrigo Cajade Jose M. Piñeiro Ana M. Torres Igor R. F. da Silva Stephen Hyslop Laura C. Leiva Daniel C. Pimenta Soledad Bustillo Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
topic_facet |
Argenteohyla siemersi Hylidae Skin secretion Tree frog Cytotoxicity Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
Abstract Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs. |
format |
Article in Journal/Newspaper |
author |
Luciano S. Fusco Rodrigo Cajade Jose M. Piñeiro Ana M. Torres Igor R. F. da Silva Stephen Hyslop Laura C. Leiva Daniel C. Pimenta Soledad Bustillo |
author_facet |
Luciano S. Fusco Rodrigo Cajade Jose M. Piñeiro Ana M. Torres Igor R. F. da Silva Stephen Hyslop Laura C. Leiva Daniel C. Pimenta Soledad Bustillo |
author_sort |
Luciano S. Fusco |
title |
Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
title_short |
Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
title_full |
Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
title_fullStr |
Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
title_full_unstemmed |
Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae) |
title_sort |
biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted argentina frog argenteohyla siemersi (anura: hylidae) |
publisher |
SciELO |
publishDate |
2020 |
url |
https://doi.org/10.1590/1678-9199-jvatitd-2019-0078 https://doaj.org/article/5c0e65622aa24f33b5008a09c4c24385 |
geographic |
Arctic Argentina |
geographic_facet |
Arctic Argentina |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100306&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20190078.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2019-0078 https://doaj.org/article/5c0e65622aa24f33b5008a09c4c24385 |
op_doi |
https://doi.org/10.1590/1678-9199-jvatitd-2019-0078 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
container_volume |
26 |
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1766344109220429824 |