Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy

Johannes Lørup Buch, Hans Ramløv

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Larvae of the blackspotted pliers support beetle, Rhagium mordax, were collected monthly, for the duration of 2012 and fixed. The larvae were embedded in paraffin wax and sectioned. Using fluorophore-coupled antibodies specific to the R. mordax antifreeze protein, RmAFP1, sections were visualised with UV reflected light microscopy. An automated software analysis method was developed in order to discard autofluorescence, and quantify fluorescence from bound antibodies. The results show that R. mordax cuticle and gut exhibit a higher degree of fluorophore-bound fluorescence during summer, than in the cold months. It is hypothesised that R. mordax stores RmAFP1 in, or near, the fat body during times when freeze avoidance is not needed.
OriginalsprogEngelsk
TidsskriftCryobiology
Vol/bind74
Sider (fra-til)132-140
ISSN0011-2240
DOI
StatusUdgivet - 2017

Bibliografisk note

The attached version of the article is wrongly specified as a 'Preprint'. This is the peer reviewed version.

Emneord

  • Antifreeze protein
  • immunoflourescence
  • Tissue distribution
  • Rhagium mordax

Citer dette

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title = "Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy",
abstract = "Larvae of the blackspotted pliers support beetle, Rhagium mordax, were collected monthly, for the duration of 2012 and fixed. The larvae were embedded in paraffin wax and sectioned. Using fluorophore-coupled antibodies specific to the R. mordax antifreeze protein, RmAFP1, sections were visualised with UV reflected light microscopy. An automated software analysis method was developed in order to discard autofluorescence, and quantify fluorescence from bound antibodies. The results show that R. mordax cuticle and gut exhibit a higher degree of fluorophore-bound fluorescence during summer, than in the cold months. It is hypothesised that R. mordax stores RmAFP1 in, or near, the fat body during times when freeze avoidance is not needed.",
keywords = "Antifreeze protein, immunoflourescence, Tissue distribution, Rhagium mordax",
author = "Buch, {Johannes L{\o}rup} and Hans Raml{\o}v",
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year = "2017",
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Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy. / Buch, Johannes Lørup; Ramløv, Hans.

I: Cryobiology, Bind 74, 2017, s. 132-140.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Detecting seasonal variation of antifreeze protein distribution in Rhagium mordax using immunofluorescence and high resolution microscopy

AU - Buch, Johannes Lørup

AU - Ramløv, Hans

N1 - The attached version of the article is wrongly specified as a 'Preprint'. This is the peer reviewed version.

PY - 2017

Y1 - 2017

N2 - Larvae of the blackspotted pliers support beetle, Rhagium mordax, were collected monthly, for the duration of 2012 and fixed. The larvae were embedded in paraffin wax and sectioned. Using fluorophore-coupled antibodies specific to the R. mordax antifreeze protein, RmAFP1, sections were visualised with UV reflected light microscopy. An automated software analysis method was developed in order to discard autofluorescence, and quantify fluorescence from bound antibodies. The results show that R. mordax cuticle and gut exhibit a higher degree of fluorophore-bound fluorescence during summer, than in the cold months. It is hypothesised that R. mordax stores RmAFP1 in, or near, the fat body during times when freeze avoidance is not needed.

AB - Larvae of the blackspotted pliers support beetle, Rhagium mordax, were collected monthly, for the duration of 2012 and fixed. The larvae were embedded in paraffin wax and sectioned. Using fluorophore-coupled antibodies specific to the R. mordax antifreeze protein, RmAFP1, sections were visualised with UV reflected light microscopy. An automated software analysis method was developed in order to discard autofluorescence, and quantify fluorescence from bound antibodies. The results show that R. mordax cuticle and gut exhibit a higher degree of fluorophore-bound fluorescence during summer, than in the cold months. It is hypothesised that R. mordax stores RmAFP1 in, or near, the fat body during times when freeze avoidance is not needed.

KW - Antifreeze protein

KW - immunoflourescence

KW - Tissue distribution

KW - Rhagium mordax

U2 - 10.1016/j.cryobiol.2016.11.003

DO - 10.1016/j.cryobiol.2016.11.003

M3 - Journal article

VL - 74

SP - 132

EP - 140

JO - Cryobiology

JF - Cryobiology

SN - 0011-2240

ER -