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Expression analysis and identification of antimicrobial peptide transcripts from six North American frog species

Diseases of Aquatic Organisms

By:
, , ,
DOI: 10.3354/dao02601

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Abstract

Frogs secrete antimicrobial peptides onto their skin. We describe an assay to preserve and analyze antimicrobial peptide transcripts from field-collected skin secretions that will complement existing methods for peptide analysis. We collected skin secretions from 4 North American species in the field in California and 2 species in the laboratory. Most frogs appeared healthy after release; however, Rana boylii in the Sierra Nevada foothills, but not the Coast Range, showed signs of morbidity and 2 died after handling. The amount of total RNA extracted from skin secretions was higher in R. boylii and R. sierrae compared to R. draytonii, and much higher compared to Pseudacris regilla. Interspecies variation in amount of RNA extracted was not explained by size, but for P. regilla it depended upon collection site and date. RNA extracted from skin secretions from frogs handled with bare hands had poor quality compared to frogs handled with gloves or plastic bags. Thirty-four putative antimicrobial peptide precursor transcripts were identified. This study demonstrates that RNA extracted from skin secretions collected in the field is of high quality suitable for use in sequencing or quantitative PCR (qPCR). However, some species do not secrete profusely, resulting in very little extracted RNA. The ability to measure transcript abundance of antimicrobial peptides in field-collected skin secretions complements proteomic analyses and may provide insight into transcriptional mechanisms that could affect peptide abundance.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
Expression analysis and identification of antimicrobial peptide transcripts from six North American frog species
Series title:
Diseases of Aquatic Organisms
DOI:
10.3354/dao02601
Volume
104
Issue:
3
Year Published:
2013
Language:
English
Publisher:
Inter-Research
Contributing office(s):
Leetown Science Center
Description:
12 p.
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
Larger Work Title:
Diseases of Aquatic Organisms
First page:
225
Last page:
236