Cargando…

Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers

Determining the age of juvenile blow flies is one of the key tasks of forensic entomology when providing evidence for the minimum post mortem interval. While the age determination of blow fly larvae is well established using morphological parameters, the current study focuses on molecular methods fo...

Descripción completa

Detalles Bibliográficos
Autores principales: Zajac, Barbara K., Amendt, Jens, Verhoff, Marcel A., Zehner, Richard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5867874/
https://www.ncbi.nlm.nih.gov/pubmed/29522463
http://dx.doi.org/10.3390/genes9030153
_version_ 1783309042724110336
author Zajac, Barbara K.
Amendt, Jens
Verhoff, Marcel A.
Zehner, Richard
author_facet Zajac, Barbara K.
Amendt, Jens
Verhoff, Marcel A.
Zehner, Richard
author_sort Zajac, Barbara K.
collection PubMed
description Determining the age of juvenile blow flies is one of the key tasks of forensic entomology when providing evidence for the minimum post mortem interval. While the age determination of blow fly larvae is well established using morphological parameters, the current study focuses on molecular methods for estimating the age of blow flies during the metamorphosis in the pupal stage, which lasts about half the total juvenile development. It has already been demonstrated in several studies that the intraspecific variance in expression of so far used genes in blow flies is often too high to assign a certain expression level to a distinct age, leading to an inaccurate prediction. To overcome this problem, we previously identified new markers, which show a very sharp age dependent expression course during pupal development of the forensically-important blow fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) by analyzing massive parallel sequencing (MPS) generated transcriptome data. We initially designed and validated two quantitative polymerase chain reaction (qPCR) assays for each of 15 defined pupal ages representing a daily progress during the total pupal development if grown at 17 °C. We also investigated whether the performance of these assays is affected by the ambient temperature, when rearing pupae of C. vicina at three different constant temperatures—namely 17 °C, 20 °C and 25 °C. A temperature dependency of the performance could not be observed, except for one marker. Hence, for each of the defined development landmarks, we can present gene expression profiles of one to two markers defining the mentioned progress in development.
format Online
Article
Text
id pubmed-5867874
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-58678742018-03-27 Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers Zajac, Barbara K. Amendt, Jens Verhoff, Marcel A. Zehner, Richard Genes (Basel) Article Determining the age of juvenile blow flies is one of the key tasks of forensic entomology when providing evidence for the minimum post mortem interval. While the age determination of blow fly larvae is well established using morphological parameters, the current study focuses on molecular methods for estimating the age of blow flies during the metamorphosis in the pupal stage, which lasts about half the total juvenile development. It has already been demonstrated in several studies that the intraspecific variance in expression of so far used genes in blow flies is often too high to assign a certain expression level to a distinct age, leading to an inaccurate prediction. To overcome this problem, we previously identified new markers, which show a very sharp age dependent expression course during pupal development of the forensically-important blow fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) by analyzing massive parallel sequencing (MPS) generated transcriptome data. We initially designed and validated two quantitative polymerase chain reaction (qPCR) assays for each of 15 defined pupal ages representing a daily progress during the total pupal development if grown at 17 °C. We also investigated whether the performance of these assays is affected by the ambient temperature, when rearing pupae of C. vicina at three different constant temperatures—namely 17 °C, 20 °C and 25 °C. A temperature dependency of the performance could not be observed, except for one marker. Hence, for each of the defined development landmarks, we can present gene expression profiles of one to two markers defining the mentioned progress in development. MDPI 2018-03-09 /pmc/articles/PMC5867874/ /pubmed/29522463 http://dx.doi.org/10.3390/genes9030153 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zajac, Barbara K.
Amendt, Jens
Verhoff, Marcel A.
Zehner, Richard
Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title_full Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title_fullStr Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title_full_unstemmed Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title_short Dating Pupae of the Blow Fly Calliphora vicina Robineau–Desvoidy 1830 (Diptera: Calliphoridae) for Post Mortem Interval—Estimation: Validation of Molecular Age Markers
title_sort dating pupae of the blow fly calliphora vicina robineau–desvoidy 1830 (diptera: calliphoridae) for post mortem interval—estimation: validation of molecular age markers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5867874/
https://www.ncbi.nlm.nih.gov/pubmed/29522463
http://dx.doi.org/10.3390/genes9030153
work_keys_str_mv AT zajacbarbarak datingpupaeoftheblowflycalliphoravicinarobineaudesvoidy1830dipteracalliphoridaeforpostmortemintervalestimationvalidationofmolecularagemarkers
AT amendtjens datingpupaeoftheblowflycalliphoravicinarobineaudesvoidy1830dipteracalliphoridaeforpostmortemintervalestimationvalidationofmolecularagemarkers
AT verhoffmarcela datingpupaeoftheblowflycalliphoravicinarobineaudesvoidy1830dipteracalliphoridaeforpostmortemintervalestimationvalidationofmolecularagemarkers
AT zehnerrichard datingpupaeoftheblowflycalliphoravicinarobineaudesvoidy1830dipteracalliphoridaeforpostmortemintervalestimationvalidationofmolecularagemarkers