Fremdt Heike, Amendt Jens, Zehner Richard
Institute of Forensic Medicine, Goethe-University Frankfurt, Kennedyallee 104, 60596, Frankfurt, Germany,
Int J Legal Med. 2014 Nov;128(6):1001-11. doi: 10.1007/s00414-013-0920-x. Epub 2013 Sep 29.
Estimating the post mortem interval (PMImin) by age determination of blow fly larvae has been well-established for moderate temperatures. Low-temperature developmental data is only available sparsely and usually does not take overwintering strategies into account. The blow fly Calliphora vicina hibernates by diapausing in the third larval stage extending the duration of this developmental stage up to several weeks or even months. As the diagnosis of the diapause status is not possible by morphological characteristics, PMImin estimations might be biased during the cold season if only based on age determination of third instar larvae of C. vicina. Molecular markers were searched for which allows one to identify diapause in larvae. Expression analysis of 19 genes was performed in diapausing and non-diapausing larvae. Three genes encoding for heat shock proteins (hsp23, hsp24 and hsp70) were found to be up-regulated distinctly in diapausing larvae and at 1 day in non-diapausing larvae. If several larvae are subjected to an analysis, a high variance in the expression level of the gene encoding for the anterior fat body protein is a further marker for diapause. The present study proves the potential use of gene expression analysis as a suitable diagnosis tool for diapause in C. vicina.
通过确定丽蝇幼虫的年龄来估计死后间隔时间(PMImin)在中等温度条件下已得到充分确立。低温发育数据非常稀少,而且通常没有考虑越冬策略。红头丽蝇通过在第三幼虫阶段滞育来冬眠,这使得该发育阶段的持续时间延长至数周甚至数月。由于无法通过形态特征诊断滞育状态,因此在寒冷季节,如果仅基于红头丽蝇三龄幼虫的年龄确定来估计PMImin,可能会产生偏差。人们寻找了能够识别幼虫滞育的分子标记。对滞育和非滞育幼虫进行了19个基因的表达分析。发现三个编码热休克蛋白的基因(hsp23、hsp24和hsp70)在滞育幼虫中明显上调,在非滞育幼虫中在第1天上调。如果对多个幼虫进行分析,编码前脂肪体蛋白的基因表达水平的高变异性是滞育的另一个标记。本研究证明了基因表达分析作为红头丽蝇滞育合适诊断工具的潜在用途。