家蚕snoRNA Bm-15的细胞定位及与类Notch受体基因的互作关系

doi:10.16380/j.kcxb.2019.08.001

摘要/Abstract

摘要：

【目的】前期研究中发现的一个家蚕Bombyx mori C/D box snoRNA Bm-15能够与类Notch受体基因(Notch-like receptor gene, NLR)发生体外互作。本研究旨在了解家蚕中snoRNA Bm-15与NLR的互作机制。【方法】利用实时荧光定量PCR检测snoRNA Bm-15和NLR在家蚕幼虫、蛹及成虫不同发育阶段的表达谱；利用原位杂交鉴定snoRNA Bm-15与NLR在BmN4细胞中的定位，以探索两者之间的互作机制。【结果】表达谱分析发现，snoRNA Bm-15及NLR在家蚕幼虫期的表达趋势基本一致，均呈现先下降后上升的趋势，2龄幼虫中表达量最低。在家蚕化蛹过程中，snoRNA Bm-15表达量呈现逐渐下降的趋势，到处女蛾时表达量最低。而NLR在雌虫化蛹后第1天表达量最低，处女蛾中表达量最高。利用原位杂交进行细胞定位发现，snoRNA Bm-15不仅存在于BmN4细胞核中，在细胞质中也有分布；而NLR则大部分分布于细胞核中，与Bm-15在核内有位置重叠。【结论】首次在鳞翅目昆虫中发现一个snoRNA Bm-15同时存在于细胞核和细胞质中，与其互作的基因NLR则主要存在于细胞核中，与Bm-15有位置重叠。二者在家蚕幼虫阶段存在一致的表达模式，在化蛹及化蛾过程中存在相反的表达模式，提示snoRNA Bm-15与NLR在家蚕不同发育阶段可能存在不同的互作模式。

关键词: 家蚕, 发育, snoRNA, Bm-15, 表达谱, 细胞定位

Abstract: 【Aim】 In a previous study, we found that a C/D box snoRNA Bm-15 of the silkworm, Bombyx mori, could interact with the Notch-like receptor gene NLR in vitro. This study aims to further investigate the mechanisms of interaction between snoRNA Bm-15 and NLR in B. mori. 【Methods】 The expression profiles of snoRNA Bm-15 and NLR in B. mori at different developmental stages of larva, pupa and adult were detected by real-time quantitative PCR. Meanwhile, the cellular location of snoRNA Bm-15 and NLR was detected by in situ hybridization in BmN4 cells to explore the interaction between them. 【Results】 Expression profiles showed that the expression trends of snoRNA Bm-15 and NLR were consistent during larval development of the silkworm, decreasing first and then increasing, with the lowest expression level in the 2nd instar larvae. During pupation and eclosion, the expression level of snoRNA Bm-15 gradually declined, with the lowest expression level in virgin moths. But NLR showed the lowest expression level in the 1 day-old female pupae and the highest expression level in virgin moths. Cellular localization revealed that snoRNA Bm-15 was not only located in the nucleus of BmN4 cells but also in the cytoplasm, while NLR was distributed mainly in the nucleus with overlap with snoRNA Bm-15. 【Conclusion】 This is the first report that a snoRNA Bm-15 exists in both nucleus and cytoplasm in lepidopteran insects, and its interacting gene NLR is mainly in the nucleus and overlapped with snoRNA Bm-15. snoRNA Bm-15 and NLR have consistent expression patterns in the larval stage of the silkworm, but show opposite expression patterns during pupation and eclosion, suggesting that snoRNA Bm-15 and NLR might have different interaction modes in different developmental stages of the silkworm.

Key words: Bombyx mori, development, snoRNA, Bm-15, expression profile, cellular location

申雅文, 马田田, 霍春月, 杨宗霖, 路一平, 阚云超, 李丹丹. 家蚕snoRNA Bm-15的细胞定位及与类Notch受体基因的互作关系[J]. 昆虫学报, 2019, 62(8): 895-900.

SHEN Ya-Wen, MA Tian-Tian, HUO Chun-Yue, YANG Zong-Lin, LU Yi-Ping, KAN Yun-Chao, LI Dan-Dan. Cellular location of snoRNA Bm-15 and its interaction with Notch-like receptor gene in the silkworm, Bombyx mori[J]. Acta Entomologica Sinica, 2019, 62(8): 895-900.

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家蚕snoRNA Bm-15的细胞定位及与类Notch受体基因的互作关系

Cellular location of snoRNA Bm-15 and its interaction with Notch-like receptor gene in the silkworm, Bombyx mori

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