Sex Genotyping of Archival Fixed and Immunolabeled Guinea Pig Cochleas
Sex Genotyping of Archival Fixed and Immunolabeled Guinea Pig Cochleas
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DOI：
10.1038/s41598-018-23491-3
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发表时间：
2018-03-26
影响因子：
4.6
通讯作者：
Whitlon, Donna S.
中科院分区：
文献类型：
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作者：
Depreux, Frederic F.;Czech, Lyubov;Whitlon, Donna S.
几十年来，远交豚鼠(GP)一直被用作研究模型。过去使用豚鼠进行的各种研究使用的测量方法，当时还不为人所知，可能与性别有关，但今天，档案组织可能是仅存的。我们的目标是提供一种对档案豚鼠组织进行性别分型的方案，从而可以重新评估过去的实验对性别的影响。目前还没有适用于全科医生的PCR性基因分型方案。我们发现，已发表的GP Sry基因序列与两个不同的GP群体中的序列不同。我们利用其他物种的序列，推导出Sry的聚合酶链式反应引物。在开发了一种用于存档、固定、脱钙、免疫标记的豚鼠耳蜗半转的基因组DNA提取方法后，我们使用了多重分析(Y特异性Sry；X特异性Dstrophin)来对3岁大的组织进行性别鉴定。这一程序应该允许重新评估先前在不同研究领域的豚鼠研究性别对实验结果的影响。
For decades, outbred guinea pigs (GP) have been used as research models. Various past research studies using guinea pigs used measures that, unknown at the time, may be sex-dependent, but from which today, archival tissues may be all that remain. We aimed to provide a protocol for sex-typing archival guinea pig tissue, whereby past experiments could be re-evaluated for sex effects. No PCR sex-genotyping protocols existed for GP. We found that published sequence of the GP Sry gene differed from that in two separate GP stocks. We used sequences from other species to deduce PCR primers for Sry. After developing a genomic DNA extraction for archival, fixed, decalcified, immunolabeled, guinea pig cochlear half-turns, we used a multiplex assay (Y-specific Sry; X-specific Dystrophin) to assign sex to tissue as old as 3 years. This procedure should allow reevaluation of prior guinea pig studies in various research areas for the effects of sex on experimental outcomes.