A Rapid, Direct Assay to Measure Degranulation of Primary Granules in Neutrophils from Kidney of Fathead Minnow (Pimephales Promelas Rafinesque, 1820)

dc.contributor.author Andreasen, Claire
dc.contributor.author Andreasen, Claire
dc.contributor.author Roth, James
dc.contributor.author Roth, James
dc.contributor.department Veterinary Pathology
dc.date 2018-02-16T19:38:21.000
dc.date.accessioned 2020-07-07T05:16:03Z
dc.date.available 2020-07-07T05:16:03Z
dc.date.issued 2005-09-01
dc.description.abstract <p>A direct, rapid, quantitative colorimetric assay to determine neutrophil primary granule degranulation was adapted for use with fathead minnow kidney neutrophils. The assay measures the exocytosis of myeloperoxidase (MPO) using 3,30,5,50-tetramethylbenzidine as a substrate. The assay was validated by comparing the total myeloperoxidase content of neutrophil populations obtained from adult cattle, as a known positive, and fish; evaluating the effects of calcium ionophore (CaI), phorbol myristate acetate (PMA), aqueous solution of b-glucan (MGAQ) and zymosan (Z) with and without cytochalasin B (cyto B) as stimulants of degranulation; determining the kinetics of primary granule exocytosis and detecting changes in degranulation when fish were exposed to stress and anaesthesia with MS-222. The MPO assay detected MPO activity in fathead minnow neutrophils that correlated to neutrophil numbers, confirmed that degranulation was increased when CaI was used compared to other stimulants, determined degranulation peak at 60 min and confirmed decreased degranulation after exposure to handling and crowding stress, with and without MS-222. Therefore, the MPO assay is capable of detecting important differences that may occur in degranulation of fathead minnow kidney neutrophil primary granules and in total neutrophil myeloperoxidase content.</p>
dc.description.comments <p>This article is from <em>Fish & Shellfish Immunology</em> 19 (2005): 217, doi:<a href="http://dx.doi.org/10.1016/j.fsi.2004.12.003" target="_blank">10.1016/j.fsi.2004.12.003</a>.</p>
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dc.identifier archive/lib.dr.iastate.edu/vpath_pubs/67/
dc.identifier.articleid 1071
dc.identifier.contextkey 7356813
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath vpath_pubs/67
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/92494
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/vpath_pubs/67/2005_Andreasen_RapidDirect.pdf|||Sat Jan 15 01:26:59 UTC 2022
dc.source.uri 10.1016/j.fsi.2004.12.003
dc.subject.disciplines Natural Resources Management and Policy
dc.subject.disciplines Veterinary Pathology and Pathobiology
dc.subject.disciplines Veterinary Preventive Medicine, Epidemiology, and Public Health
dc.subject.keywords neutrophils
dc.subject.keywords fish
dc.subject.keywords degranulation assay
dc.subject.keywords fathead minnow
dc.subject.keywords Veterinary Microbiology & Preventive Medicine
dc.title A Rapid, Direct Assay to Measure Degranulation of Primary Granules in Neutrophils from Kidney of Fathead Minnow (Pimephales Promelas Rafinesque, 1820)
dc.type article
dc.type.genre article
dspace.entity.type Publication
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relation.isAuthorOfPublication 909dd0b2-ec0a-41e2-b4e0-9e5ff76b7622
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