A novel method for the release and collection of dermal, glandular secretions from the skin of frogs

Tyler, Michael J.; Stone, David J.; Bowie, John H.

doi:10.1016/1056-8719(92)90004-k

Cited by 236 publications

(141 citation statements)

References 7 publications

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“…This method was designed to provide a standardized stimulus to induce skin secretions from a variety of small tropical amphibians and to avoid the more painful transcutaneous electrostimulation procedure, which requires calibration for each species (Tyler et al, 1992). Amphibians were captured by hand inside new plastic bags and weighed within 0.1 g. With minimal handling, frogs were injected with 10 nmol (0.01 ml) per gram body weight of norepinephrine (bitartrate salt, Sigma, St. Louis, Missouri, USA).…”

Section: Collection Of Skin Peptidesmentioning

confidence: 99%

Predicted Disease Susceptibility in a Panamanian Amphibian Assemblage Based on Skin Peptide Defenses

Woodhams

Voyles

Lips

et al. 2006

Journal of Wildlife Diseases

138

139

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ABSTRACT:Chytridiomycosis is an emerging infectious disease of amphibians caused by a chytrid fungus, Batrachochytrium dendrobatidis. This panzootic does not equally affect all amphibian species within an assemblage; some populations decline, others persist. Little is known about the factors that affect disease resistance. Differences in behavior, life history, biogeography, or immune function may impact survival. We found that an innate immune defense, antimicrobial skin peptides, varied significantly among species within a rainforest stream amphibian assemblage that has not been exposed to B. dendrobatidis. If exposed, all amphibian species at this central Panamanian site are at risk of population declines. In vitro pathogen growth inhibition by peptides from Panamanian species compared with species with known resistance (Rana pipiens and Xenopus laevis) or susceptibility (Bufo boreas) suggests that of the nine species examined, two species (Centrolene prosoblepon and Phyllomedusa lemur) may demonstrate strong resistance, and the other species will have a higher risk of disease-associated population declines. We found little variation among geographically distinct B. dendrobatidis isolates in sensitivity to an amphibian skin peptide mixture. This supports the hypothesis that B. dendrobatidis is a generalist pathogen and that species possessing an innate immunologic defense at the time of disease emergence are more likely to survive.

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Section: Collection Of Skin Peptidesmentioning

confidence: 99%

Predicted Disease Susceptibility in a Panamanian Amphibian Assemblage Based on Skin Peptide Defenses

Woodhams

Voyles

Lips

et al. 2006

Journal of Wildlife Diseases

138

139

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“…Adult animals were maintained in captivity at the Biology Department of the Moscow State University under conditions close to natural ones. Secretion from the skin glands was provoked by mild electrical stimulation [19] for 40 s using laboratory electrostimulator ESL-1 equipped with platinum electrodes. The duration of impulse was 3 ms with amplitude of 10 V at 50 Hz.…”

Section: Preparation Of Skin Secretionsmentioning

confidence: 99%

Oxidation versus carboxamidomethylation of s-s bond in ranid frog peptides: Pro and contra for de novo MALDI-MS sequencing

Samgina

Artemenko

Gorshkov

et al. 2008

J. Am. Soc. Mass Spectrom.

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Five natural peptides isolated from ranid skin secretions of European frog species of Rana ridibunda and Rana arvalis (molecular masses 3516, 26743516, , 26363516, , 18743516, , and 1810 were studied by MALDI-TOF/TOF to compare two procedures of disulfide bond cleavage: (1) performic oxidation and (2) reduction/carboxamidomethylation. The processes are relevant for the elucidation of the amino acid sequence inside the seven-member cystine ring at the C-terminus. The results clearly demonstrated that oxidation of the disulfide bond led to notably higher abundances of b-and y-ions, corresponding to the C-terminal peptide bonds, than reduction/carboxamidomethylation. This conclusion is true for all five peptides studied. Besides that, the oxidation procedure is simpler than carboxamidomethylation, as it is a one-step process with no purification required. The oxidation is more reproducible. The results were similar each time the peptide was subjected to the process. It was successfully applied to all five peptides while reduction/carboxamidomethylation failed in the case of brevinin-1Ra, despite all variations of reaction conditions. (J Am Soc Mass Spectrom 2008, 19, 479 -487) © 2008 American Society for Mass Spectrometry A skin secretion produced by granular skin glands of anurans in response to a variety of stimuli contains biologically active peptides [1][2][3][4]. Our interest was focused on the Rana genus containing a notable array of common frogs spread around the territory of Eurasia and America. A major part of ranid peptides has a disulfide cycle at the C-terminus [1,5]. This peculiarity, called "Rana box" [6], complicates sequencing of these peptides. The amino acid sequence inside the cycle is practically unattainable by means of mass spectrometry without preliminary chemical derivatization. There is only one communication on direct sequence identification inside the C-terminus disulfide cycle using ESI MS/MS in the negative ion mode. It deals with some bioactive peptides isolated from genus Crinia (Australia) and with brevinin-1E R. ridibunda, (Russia) [7]. In this case, a combination of the positive and negative mass spectra has produced complete sequences for all the investigated peptides. Unfortunately not all peptides give reasonable negative ion spectra.Mass spectrometric sequencing of disulfide containing peptides requires preliminary modification of the S-S bond. There are two general methods to cleave this bond: (1) reduction followed by the alkylation of cysteines [8 -10]; (2) oxidation with performic acid [11]. The first procedure is traditionally used to break S-S bonds in proteins with associated polypeptide chains before their enzymatic digestion. We applied it for the skin peptides of the Rana genus with an internal disulfide cycle [12,13]. Reduction of the S-S bond followed by carboxamidomethylation allows reliable sequencing at the C-terminus. LTQ FTICR mass spectrometric results completely match those obtained by Edman degradation.Performic oxidation is a well-known method for the cl...

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“…ii. Physical stimuli: mild electrical stimulation has been demonstrated to be an effective method to stimulate the release of amphibian skin secretions even in species lacking enlarged compact glands (38). Another simple method to obtain amphibian skin secretions is to press the granular glands gently so as to milk skin secretions, which is suitable for amphibians with enlarged compact glands.…”

Section: Identification Of Amphibian Ampsmentioning

confidence: 99%

“…Once stimulated, secretions produced within the cell discharge by the rupture of the plasma membrane, thus releasing the cellular contents into the lumen. AMPs stored in the granular glands in prepropeptide form will remove the signal peptide from the precursor and release into the skin surface in a holocrine manner upon stress and injury (37,38).…”

Section: Expression and Post-translational Modifications Of Amphibianmentioning

confidence: 99%

Antimicrobial peptides from amphibians

Xiao

Liu

Lai

2011

BioMolecular Concepts

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Increased prevalence of multi-drug resistance in pathogens has encouraged researchers to focus on finding novel forms of anti-infective agents. Antimicrobial peptides (AMPs) found in animal secretions are components of host innate immune response and have survived eons of pathogen evolution. Thus, they are likely to be active against pathogens and even those that are resistant to conventional drugs. Many peptides have been isolated and shown to be effective against multi-drug resistant pathogens. More than 500 AMPs have been identified from amphibians. The abundance of AMPs in frog skin is remarkable and constitutes a rich source for design of novel pharmaceutical molecules. Expression and post-translational modifications, discovery, activities and probable therapeutic application prospects of amphibian AMPs will be discussed in this article.

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A novel method for the release and collection of dermal, glandular secretions from the skin of frogs

Cited by 236 publications

References 7 publications

Predicted Disease Susceptibility in a Panamanian Amphibian Assemblage Based on Skin Peptide Defenses

Predicted Disease Susceptibility in a Panamanian Amphibian Assemblage Based on Skin Peptide Defenses

Oxidation versus carboxamidomethylation of s-s bond in ranid frog peptides: Pro and contra for de novo MALDI-MS sequencing

Antimicrobial peptides from amphibians

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