|
|
|
| Myeloid and erythroid hematopoietic transcription factor expression decline after knockdown of lmna genes in zebrafish embryos |
| HOU Shifang1,2, WANG Zhihua1, WANG Jun3, HE Zhixu2,4, SHU Liping1,2,5 |
1. Department of Immunology, Guizhou Medical University, Guiyang 550004, China;
2. Tissue Engineering and Stem Cell Research Center, Guizhou Medical University, Guiyang 550004, China;
3. Clinical Medical Research Center, Guizhou Medical University, Guiyang 550004, China;
4. Children Medical Center, Guizhou Medical University, Guiyang 550004, China;
5. Laboratory Animal Center, Guizhou Medical University, Guiyang 550004, China |
|
|
|
Abstract Objective: To investigate the effect of lmna gene down-regulation on early hematopoietic development of zebrafish. Methods: Phosphorodiamidate morpholino oligomer (PMO) technology was used to downregulate lmna gene expression in Zebrafish. Zebrafish embryos injected phosphorodiamidate morpholino antisense oligonucleotide of lmna gene mRNA by microinjection at unicellular stage were taken as the experimental group, and those injected meaningless phosphorodiamidate morpholino antisense oligonucleotide were taken as the control. The embryos were collected at 18, 24, 30 and 36 hpf after the fertilization. The real-time fluorescent quantitative PCR (RT-PCR) and whole embryo in situ hybridization methods were used to detect the expression of myeloid hematopoietic transcription factor pu. 1 and erythroid hematopoietic transcription factor gata1 in zebrafish. Results: RT-PCR showed that the expressions of pu. 1 and gata1 decreased in the experimental group compared with the control group (all P<0.05). Whole embryo in situ hybridization showed that the blue-black positive hybridization signals of pu. 1 and gata1 in experimental group were shallow than those in the control group. Conclusion: Myeloid hematopoietic and erythroid hematopoietic of zebrafish are blocked with the downregulation of lmna gene.
|
|
Received: 20 August 2016
|
|
|
下调lmna基因对斑马鱼胚胎髓系和红系造血干细胞发育的影响
目的：探讨lmna基因下调对斑马鱼早期造血干细胞发育的影响。方法：显微注射法将lmna基因mRNA的吗啉代反义寡核苷酸注入单细胞期斑马鱼胚胎（实验组），显微注射无意义的吗啉代寡核苷酸作为对照组。待胚胎发育至受精18、24、30、36 hpf后收集胚胎进行实验。RT-PCR和全胚胎原位杂交方法检测斑马鱼髓系造血转录因子pu. 1和红系造血转录因子gata1的变化。结果：RT-PCR结果显示，实验组pu. 1和gata1表达均较对照组下降（均P<0.05）。原位杂交结果显示，实验组pu. 1和gata1蓝黑色阳性杂交信号较对照组变浅。结论：lmna基因下调会阻碍斑马鱼髓系造血干细胞和红系造血干细胞的发育。
关键词：
斑马鱼,
核纤层蛋白A型,
转录因子,
基因,
基因表达调控,
造血干细胞,
寡核苷酸类,
反义
|
|
| [[1]] |
WU D, FLANNERY A R, CAI H, et al. Nuclear localization signal deletion mutants of lamin A and progerin reveal insights into lamin A processing and emerin targeting[J]. Nucleus, 2014, 5(1):66-74.
|
|
|
| [[2]] |
ROCHA-PERUGINI V, GONZÁLEZ-GRANADO J M. Nuclear envelope lamin-A as a coordinator of T cell activation[J]. Nucleus, 2014, 5(5):396-401.
|
|
|
| [[3]] |
KOSHIMIZU E, IMAMURA S, QI J, et al. Embryonic senescence and laminopathies in a progeroid zebrafish model[J/OL]. PLoS One, 2011, 6(3):e17688.
|
|
|
| [[4]] |
MALASHICHEVA A B, ZABIRNIK A S, SMOLINA N A, et al. Lamin A/C mutations change differentiation potential of mesenchymal stem cells[J]. Tsitologiia, 2013, 55(5):313-317.
|
|
|
| [[5]] |
MORY P B, CRISPIM F, FREIRE M B, et al. Phenotypic diversity in patients with lipodystrophy associated with LMNA mutations[J]. Eur J Endocrinol, 2012, 167(3):423-431.
|
|
|
| [[6]] |
ZHANG L, SHEN H, ZHAO Z, et al. Cardiac effects of the c.1583 C->G LMNA mutation in two families with Emery-Dreifuss muscular dystrophy[J]. Mol Med Rep, 2015, 12(4):5065-5071.
|
|
|
| [[7]] |
MCKENNA T, ROSENGARDTEN Y, VICECONTE N, et al. Embryonic expression of the common progeroid lamin A splice mutation arrests postnatal skin development[J]. Aging Cell, 2014, 13(2):292-302.
|
|
|
| [[8]] |
MATSUMOTO A, HIEDA M, YOKOYAMA Y, et al. Global loss of a nuclear lamina component, lamin A/C, and LINC complex components SUN1, SUN2, and nesprin-2 in breast cancer[J]. Cancer Med, 2015, 4(10):1547-1557.
|
|
|
| [[9]] |
NARDELLA M, GUGLIELMI L, MUSA C, et al. Down-regulation of the Lamin A/C in neuroblastoma triggers the expansion of tumor initiating cells[J]. Oncotarget, 2015, 6(32):32821-32840.
|
|
|
| [[10]] |
GUILLY M N, KOLB J P, GOSTI F, et al. Lamins A and C are not expressed at early stages of human lymphocyte differentiation[J]. Exp Cell Res, 1990, 189(1):145-147.
|
|
|
| [[11]] |
HALE J S, FROCK R L, MAMMAN S A, et al. Cell-extrinsic defective lymphocyte development in Lmna(-/-) mice[J/OL]. PLoS One, 2010, 5(4):e10127.
|
|
|
| [[12]] |
AGRELO R, SETIEN F, ESPADA J, et al. Inactivation of the lamin A/C gene by CpG island promoter hypermethylation in hematologic malignancies, and its association with poor survival in nodal diffuse large B-cell lymphoma[J]. J Clin Oncol, 2005, 23(17):3940-3947.
|
|
|
| [[13]] |
SHU L P, ZHOU Z W, ZHOU T, et al. Ectopic expression of Hoxb4a in hemangioblasts promotes hematopoietic development in early embryogenesis of zebrafish[J]. Clin Exp Pharmacol Physiol, 2015, 42(12):1275-1286.
|
|
|
| [[14]] |
WESTERFIELD M. The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio)[M]. 4th ed. Eugene (Oregon):University of Oregon Press, 2000:2-26.
|
|
|
| [[15]] |
KIMMEL C B, BALLARD W W, KIMMEL S R, et al. Stages of embryonic development of the zebrafish[J]. Dev Dyn, 1995, 203(3):253-310.
|
|
|
| [[16]] |
AVAGYAN S, ZON L I. Fish to learn:insights into blood development and blood disorders from zebrafish hematopoiesis[J]. Hum Gene Ther, 2016, 27(4):287-294.
|
|
|
| [[17]] |
HESS I, BOEHM T. Stable multilineage xenogeneic replacement of definitive hematopoiesis in adult zebrafish[J]. Sci Rep, 2016, 6:19634.
|
|
|
| [[18]] |
RAJAN V, DELLAIRE G, BERMAN J N. Modeling Leukemogenesis in the zebrafish using genetic and xenograft models[J]. Methods Mol Biol, 2016, 1451:171-189.
|
|
|
| [[19]] |
AMANATIADOU E P, PAPADOPOULOS G L, STROUBOULIS J, et al. GATA1 and PU.1 bind to ribosomal protein genes in erythroid cells:implications for ribosomopathies[J/OL]. PLoS One, 2015, 10(10):e0140077.
|
|
|
| [[20]] |
QIAN M, JIN W, ZHU X, et al. Structurally differentiated cis-elements that interact with PU.1 are functionally distinguishable in acute promyelocytic leukemia[J]. J Hematol Oncol, 2013, 6:25.
|
|
|
| [[21]] |
SHIMIZU R, YAMAMOTO M. GATA-related hematologic disorders[J]. Exp Hematol, 2016, 44(8):696-705
|
|
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
