利用 smFISH 和原位 HCR 对 Pristionchus pacificus 的基因表达进行可视化。

microPublication biology Pub Date : 2024-07-26 eCollection Date: 2024-01-01 DOI:10.17912/micropub.biology.001274
Yasmin H Ramadan, Oliver Hobert
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引用次数: 0

摘要

单分子荧光原位杂交(smFISH)和原位杂交链反应(HCR)已成为可视化许多不同动物物种基因表达的强大工具。我们在这里展示了 smFISH 和原位 HCR 在卫星线虫模式生物 Pristionchus pacificus 中的有效应用。通过研究同源基因(Ppa-unc-30)的表达,我们发现 HCR 比 smFISH 更灵敏。我们通过检测几个参与神经递质合成或转运的基因(Ppa-unc-25 /GAD、Ppa-unc-17/VAChT、Ppa-eat-4 /VGLUT)的表达,证实了 HCR 的稳健性。与 smFISH 分析相比,原位 HCR 具有相对的成本效益,是太平洋鼠研究工具箱的有益补充。
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Visualization of gene expression in Pristionchus pacificus with smFISH and in situ HCR.

Single molecule fluorescence in situ hybridization (smFISH) and in situ hybridization chain reaction (HCR) have become powerful tools to visualize gene expression in many different animal species. We show here that smFISH and in situ HCR can be put to effective use in the satellite nematode model organism Pristionchus pacificus . Examining the expression of a homeobox gene ( Ppa-unc-30) , we found that HCR is more sensitive than smFISH. We confirmed the robustness of HCR by visualization of the expression of several genes involved in neurotransmitter synthesis or transport ( Ppa-unc-25 /GAD, Ppa-unc-17/VAChT, Ppa-eat-4 /VGLUT). Combined with its relative cost-effectiveness compared to smFISH analysis, in situ HCR constitutes a useful addition to the toolbox for P. pacificus research .

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