三倍体雌性虹鳟性腺发育过程中lc3b 基因的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2016-1166

生物技术通报 ›› 2017, Vol. 33 ›› Issue (7): 179-184.doi: 10.13560/j.cnki.biotech.bull.1985.2016-1166

三倍体雌性虹鳟性腺发育过程中lc3b 基因的表达分析

黄天晴,王星然,王彦娜,孙慧智,韩英

东北农业大学动物科学技术学院，哈尔滨 150030

收稿日期:2016-12-24 出版日期:2017-07-11 发布日期:2017-07-11
作者简介:黄天晴，女，博士研究生，研究方向：水产动物育种与繁殖；E-mail：tianqing88@126.com
基金资助:
国家自然科学基金项目（31470131）

Expression Analysis of lc3b Gene in Gonadal Development of Triploid Female Rainbow Trout

HUANG Tian-qing, WANG Xing-ran ,WANG Yan-na, SUN Hui-zhi, HAN Ying

Department of Animal Science and Technology，Northeast Agricultural University，Harbin 150030

Received:2016-12-24 Published:2017-07-11 Online:2017-07-11

摘要/Abstract

摘要： 旨在证明细胞自噬与三倍体雌性虹鳟性腺败育的关系。利用RT-PCR 扩增虹鳟自噬基因lc3b（Microtubule associated protein 1 light chain 3 beta，lc3b）的编码区序列，通过透射电镜观察三倍体雌性虹鳟不同发育阶段性腺中自噬小体的形态，结合不同发育阶段性腺中lc3b 基因和蛋白的表达，来证明细胞自噬的发生。结果表明，在200 dpf-270 dpf（Days post fertilization，dpf）这个发育阶段，lc3b 基因以及自噬蛋白LC3B-II / LC3B-I的表达处于相对较高的水平，在这个时期内，都观察到了自噬体结构的存在。这些结果证明了性腺细胞自噬为三倍体雌性虹鳟性腺败育的主要原因之一。

关键词: 三倍体, 虹鳟, 自噬, LC3B, 性腺发育

Abstract: This work aims to show a new light on the relationship between autophagy and gonadal regression of triploid female rainbow trout. Autophagy gene lc3b of rainbow trout was cloned by RT-PCR，and its expressions in gonads of triploid female rainbow trout were analyzed at different developmental period in mRNA and protein level. Gonadal ultrastructures were observed under transmission electron microscopy. Results showed clear evidence that lc3b gene was highly expressed in gonads of triploid female rainbow trout during the period of 200dpf to 270 dpf，in which autophagosome structures were identified. In this stage，the conversion of LC3B-I to LC3B-II was greater. These results demonstrated autophagy played pivotal roles in gonadal regression of triploid female rainbow trout.

Key words: triploid, rainbow trout, autophagy, LC3B, gonadal development

黄天晴,王星然,王彦娜,孙慧智,韩英. 三倍体雌性虹鳟性腺发育过程中lc3b 基因的表达分析[J]. 生物技术通报, 2017, 33(7): 179-184.

HUANG Tian-qing, WANG Xing-ran ,WANG Yan-na, SUN Hui-zhi, HAN Ying. Expression Analysis of lc3b Gene in Gonadal Development of Triploid Female Rainbow Trout[J]. Biotechnology Bulletin, 2017, 33(7): 179-184.

参考文献

[1] Xu G, Huang T, Jin X, et al. Morphology, sex steroid level andgene expression analysis in gonadal sex reversal of triploid female（XXX）rainbow trout（Oncorhynchus mykiss）[J] . Fish Physiol Biochem, 2016, 42（1）：193-202.
[2] Fader CM, Colombo MI. Autophagy and multivesicular bodies：two closely related partners[J] . Cell Death Differ, 2009, 16（1）：70-78.
[3] Degterev A, Yuan J. Expansion and evolution of cell death programmes[J] . Nat Rev Mol Cell Biol, 2008, 9（5）：378-390.
[4] Maiuri MC, Zalckvar E, Kimchi A, et al. Self-eating and self-killing：crosstalk between autophagy and apoptosis[J] . Nat Rev Mol Cell Biol, 2007, 8（9）：741-752.
[5] Mizushima N, Levine B, Cuervo A, et al. Autophagy fights disease through cellular self-digestion[J] . Nature, 2008, 451（7182）：1069-1075.
[6] Klionsky DJ. Autophagy revisited：a conversation with Christian de Duve[J] . Autophagy, 2008, 4（6）：740-743.
[7] Tanida I. Autophagy basics[J] . Microbiology and Immunology, 2011, 55（1）：1-11.
[8] Klionsky DJ, Cregg JM, Dunn WA, et al. A unified nomenclature for yeast autophagy-related genes[J] . Dev Cell, 2003, 5（4）：539- 545.
[9] Morais RDVS, Thome RG, Lemos FS, et al. Relationship between bcl-2, bax, beclin-1, and cathepsin-D proteins during postovulatory follicular regression in fish ovary[J] . Theriogenology, 2016, 85（6）：1118-1131.
[10] Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing[J] . EMBO J, 2000, 19（21）：5720-5728.
[11] Ryter SW, Mizumura K, Choi AM. The impact of autophagy on cell death modalities[J] . Int J Cell Biol, 2014, 2014：502676.
[12] Mizushima N, Yoshimori T. HowtointerpretLC 3 immunoblotting[J] . Autophagy, 2007, 3（6）：542-545.
[13] Espinosa E, Josa A, Gil L, et al. Triploidy in rainbow trout determi-ned by computer-assisted analysis[J] . J Exp Zool A Comp Exp Biol, 2005, 303（11）：1007-1012.
[14] Klionsky DJ. Autophagy：from phenomenology to molecular understanding in less than a decade[J] . Nature Reviews Molecular Cell Biology, 2007, 8（11）：931-937.
[15] Depiereux S, Liagre M, Danis L, et al. Intersex occurrence in rainbow trout（Oncorhynchus mykiss）male fry chronically exposed to ethynylestradiol[J] . PLoS One, 2014, 9（7）：e98531.
[16] Melo RMC, Martins YS, Luz RK, et al. PCNA and apoptosis during post-spawning ovarian remodeling in the teleost Oreochromis niloticus[J] . Tissue & Cell, 2015, 47（6）：541-549.
[17] Han Y, Liu M, Zhang LL, et al. Comparison of reproductive development in triploid and diploid female rainbow trout Oncorhynchus mykiss[J] . J Fish Biol, 2010, 76（7）：1742- 1750.
[18] Thome RG, Santos HB, Arantes FP, et al. Dual roles for autophagy during follicular atresia in fish ovary[J] . Autophagy, 2009, 5（1）：117-119.
[19] Sandri,M.Basicsofautophagyinmuscle-Cellularmechanisms[J] . Neuromuscular Disorders, 2014, 24（9-10）：791.
[20] Zhang Z, Singh R, Aschner M. Methods for the detection of autophagy in mammalian cells[J] . Curr Protoc Toxicol, 2016, 69：20121-201226.

三倍体雌性虹鳟性腺发育过程中lc3b 基因的表达分析

Expression Analysis of lc3b Gene in Gonadal Development of Triploid Female Rainbow Trout

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	李豫, 陈刚, 马骞, 邝杰华, 陈有铭. 军曹鱼nanos1的克隆及其在胚胎和性腺发育过程中的表达分析[J]. 生物技术通报, 2022, 38(5): 175-182.
[2]	汤晓丽, 姜福东, 张洪霞. 植物SINA E3泛素连接酶功能的研究进展[J]. 生物技术通报, 2022, 38(10): 10-17.
[3]	邹坤, 路丽丽, Collins Asiamah Amponsah, 薛缘, 张少伟, 苏瑛, 赵志辉. 家禽卵泡闭锁机制的研究进展[J]. 生物技术通报, 2020, 36(4): 185-191.
[4]	李平, 张桂萍 ,胡建燃. 连翘总黄酮对胃癌细胞MGC80-3增殖的影响[J]. 生物技术通报, 2018, 34(6): 199-203.
[5]	张加姿,李开心,于宝佳,岳洁瑜,王华忠. 小麦自噬相关因子ATG5的原核表达和抗血清制备[J]. 生物技术通报, 2017, 33(7): 69-74.
[6]	赵紫霞, 徐桂彩, 李炯棠, 杨世勇. 用于鉴别大西洋鲑和虹鳟的生物传感检测方法[J]. 生物技术通报, 2017, 33(6): 54-61.
[7]	韩坤煌, 周鹏, 邹志华, 张子平, 王艺磊. 大黄鱼Lc-UBE2D4基因的克隆及其表达分析[J]. 生物技术通报, 2017, 33(11): 166-173.
[8]	王琪, 齐仁立, 王敬, 黄金秀. 氨基酸缺乏诱导细胞自噬及miRNA调控机制[J]. 生物技术通报, 2016, 32(9): 38-43.
[9]	蒋骄云,田文斐,冯龙,曲宪成. 地高辛标记的黄鳝F64基因cRNA探针的制备及其应用[J]. 生物技术通报, 2016, 32(3): 137-141.
[10]	韩坤煌, 戴燕彬, 邹志华, 张子平, 王艺磊. CKS1B参与拟穴青蟹性腺发育的研究[J]. 生物技术通报, 2016, 32(11): 188-193.
[11]	王志舒, 谭晓荣, 刘洹洹. 线粒体自噬调控机制研究进展[J]. 生物技术通报, 2015, 31(6): 42-47.
[12]	韩冰, 杨琴, 崔清华. 斯达油脂酵母中自噬与油脂积累的关联性分析[J]. 生物技术通报, 2015, 31(2): 208-216.
[13]	王棋文,宋德荣,刘其昌,周大荣,彭华,张琼娣,罗耀. 镉诱导细胞自噬的研究进展[J]. 生物技术通报, 2015, 31(10): 56-61.
[14]	胡丹丹,刘哲,邵淑娟,杨娟,王建福. 虹鳟MHC-UBA基因遗传多样性分析[J]. 生物技术通报, 2013, 0(7): 99-106.
[15]	周广舟;张璐;卢延克;司灿;徐苏丽;. 鱼类细胞自噬研究进展[J]. , 2012, 0(04): 41-44.