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Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)

A novel antibody-based protein microarray was developed that simultaneously determines expression of 31 stress-associated proteins in skin samples collected from free-ranging grizzly bears (Ursus arctos) in Alberta, Canada. The microarray determines proteins belonging to four broad functional catego...

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Autores principales: Carlson, Ruth I., Cattet, Marc R. L., Sarauer, Bryan L., Nielsen, Scott E., Boulanger, John, Stenhouse, Gordon B., Janz, David M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4771112/
https://www.ncbi.nlm.nih.gov/pubmed/27293753
http://dx.doi.org/10.1093/conphys/cow001
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author Carlson, Ruth I.
Cattet, Marc R. L.
Sarauer, Bryan L.
Nielsen, Scott E.
Boulanger, John
Stenhouse, Gordon B.
Janz, David M.
author_facet Carlson, Ruth I.
Cattet, Marc R. L.
Sarauer, Bryan L.
Nielsen, Scott E.
Boulanger, John
Stenhouse, Gordon B.
Janz, David M.
author_sort Carlson, Ruth I.
collection PubMed
description A novel antibody-based protein microarray was developed that simultaneously determines expression of 31 stress-associated proteins in skin samples collected from free-ranging grizzly bears (Ursus arctos) in Alberta, Canada. The microarray determines proteins belonging to four broad functional categories associated with stress physiology: hypothalamic–pituitary–adrenal axis proteins, apoptosis/cell cycle proteins, cellular stress/proteotoxicity proteins and oxidative stress/inflammation proteins. Small skin samples (50–100 mg) were collected from captured bears using biopsy punches. Proteins were isolated and labelled with fluorescent dyes, with labelled protein homogenates loaded onto microarrays to hybridize with antibodies. Relative protein expression was determined by comparison with a pooled standard skin sample. The assay was sensitive, requiring 80 µg of protein per sample to be run in triplicate on the microarray. Intra-array and inter-array coefficients of variation for individual proteins were generally <10 and <15%, respectively. With one exception, there were no significant differences in protein expression among skin samples collected from the neck, forelimb, hindlimb and ear in a subsample of n = 4 bears. This suggests that remotely delivered biopsy darts could be used in future sampling. Using generalized linear mixed models, certain proteins within each functional category demonstrated altered expression with respect to differences in year, season, geographical sampling location within Alberta and bear biological parameters, suggesting that these general variables may influence expression of specific proteins in the microarray. Our goal is to apply the protein microarray as a conservation physiology tool that can detect, evaluate and monitor physiological stress in grizzly bears and other species at risk over time in response to environmental change.
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spelling pubmed-47711122016-06-10 Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos) Carlson, Ruth I. Cattet, Marc R. L. Sarauer, Bryan L. Nielsen, Scott E. Boulanger, John Stenhouse, Gordon B. Janz, David M. Conserv Physiol Toolbox A novel antibody-based protein microarray was developed that simultaneously determines expression of 31 stress-associated proteins in skin samples collected from free-ranging grizzly bears (Ursus arctos) in Alberta, Canada. The microarray determines proteins belonging to four broad functional categories associated with stress physiology: hypothalamic–pituitary–adrenal axis proteins, apoptosis/cell cycle proteins, cellular stress/proteotoxicity proteins and oxidative stress/inflammation proteins. Small skin samples (50–100 mg) were collected from captured bears using biopsy punches. Proteins were isolated and labelled with fluorescent dyes, with labelled protein homogenates loaded onto microarrays to hybridize with antibodies. Relative protein expression was determined by comparison with a pooled standard skin sample. The assay was sensitive, requiring 80 µg of protein per sample to be run in triplicate on the microarray. Intra-array and inter-array coefficients of variation for individual proteins were generally <10 and <15%, respectively. With one exception, there were no significant differences in protein expression among skin samples collected from the neck, forelimb, hindlimb and ear in a subsample of n = 4 bears. This suggests that remotely delivered biopsy darts could be used in future sampling. Using generalized linear mixed models, certain proteins within each functional category demonstrated altered expression with respect to differences in year, season, geographical sampling location within Alberta and bear biological parameters, suggesting that these general variables may influence expression of specific proteins in the microarray. Our goal is to apply the protein microarray as a conservation physiology tool that can detect, evaluate and monitor physiological stress in grizzly bears and other species at risk over time in response to environmental change. Oxford University Press 2016-02-26 /pmc/articles/PMC4771112/ /pubmed/27293753 http://dx.doi.org/10.1093/conphys/cow001 Text en © The Author 2016. Published by Oxford University Press and the Society for Experimental Biology. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Toolbox
Carlson, Ruth I.
Cattet, Marc R. L.
Sarauer, Bryan L.
Nielsen, Scott E.
Boulanger, John
Stenhouse, Gordon B.
Janz, David M.
Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title_full Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title_fullStr Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title_full_unstemmed Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title_short Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos)
title_sort development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (ursus arctos)
topic Toolbox
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4771112/
https://www.ncbi.nlm.nih.gov/pubmed/27293753
http://dx.doi.org/10.1093/conphys/cow001
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