Long Ren, Luo Chengyong, Zhang Peng, Zou Xing, Song Tao
Key Laboratory of Tropical Translational Medicine of Ministry of Education, School of Basic Medicine and Life Sciences, Hainan Medical University, Haikou, China.
Guangdong Nantian Institute of Forensic Science, Shenzhen, China.
Forensic Sci Res. 2025 Jan 16;10(3):owaf003. doi: 10.1093/fsr/owaf003. eCollection 2025 Sep.
Age estimation is a critical aspect of forensic entomology, especially in the examination of pupae. The use of proteins as a means for age identification shows great promise. In this study, proteomic techniques were employed to investigate differentially expressed proteins (DEPs) during the intrapuparial stage of . Specimens were sampled at four distinct time points: 0 h (Group A), 24 h (Group B), 48 h (Group C), and 72 h (Group D). Our analysis uncovered 56 DEPs between Groups B and A, 116 DEPs between Groups C and A, and a total of 152 DEPs between Groups D and A. These DEPs were categorized into nine clusters based on their expression patterns. Cluster 1 exhibited an increasing trend in protein expression, while Cluster 4 displayed the opposite pattern. Clusters 2, 6, and 9 showed an initial rise followed by a decline, whereas Cluster 3 demonstrated the reverse trend. Cluster 8 indicated an initial rise, a subsequent drop, and another rise, while Cluster 7 showed an initial decrease, followed by an increase and a minor decrease. Notably, the C-type lectin domain-containing (CTLD) protein and Failed axon connections (Fax) protein consistently displayed an upward trend. These two DEPs were selected for validation using the parallel reaction monitoring technique-targeted proteomics, confirming the trends observed in the initial analysis. In summary, this study highlights the potential of using proteins as reliable biomarkers for estimating pupal age.
年龄估计是法医昆虫学的一个关键方面，尤其是在蛹的检验中。利用蛋白质作为年龄鉴定手段显示出巨大潜力。在本研究中，采用蛋白质组学技术研究了[昆虫名称]蛹期内差异表达蛋白（DEPs）。在四个不同时间点采集样本：0小时（A组）、24小时（B组）、48小时（C组）和72小时（D组）。我们的分析发现，B组和A组之间有56个DEPs，C组和A组之间有116个DEPs，D组和A组之间共有152个DEPs。这些DEPs根据其表达模式被分为九个簇。簇1的蛋白质表达呈上升趋势，而簇4则呈现相反模式。簇2、6和9显示先上升后下降，而簇3呈现相反趋势。簇8显示先上升、随后下降、再上升，而簇7显示先下降、接着上升、再轻微下降。值得注意的是，含C型凝集素结构域（CTLD）蛋白和轴突连接失败（Fax）蛋白始终呈上升趋势。选择这两个DEPs使用平行反应监测技术——靶向蛋白质组学进行验证，证实了初步分析中观察到的趋势。总之，本研究突出了利用蛋白质作为估计蛹龄可靠生物标志物的潜力。
