Journal of Shandong University (Health Sciences) ›› 2019, Vol. 57 ›› Issue (5): 67-73.doi: 10.6040/j.issn.1671-7554.0.2018.1486

Previous Articles    

Effects of lentivirus injection via testicular efferent tubules on the reproductive system in Wistar rats

DI Yu, CHEN Dongshan, YAN Lei   

  1. Department of Urology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Published:2022-09-27

PDF (PC)

41

Abstract: Objective To investigate the effect of stably transgenic technology of highly toxic lentivirus on the reproductive system of male Wistar rats. Methods The stable interference lentivirus of PAX2 containing green fluorescent protein were constructed. The solution was labeled with Trypan blue. The testicular efferent tubules injections of 16 Wistar rats were carried out with a 34 G needle. The testis parenchyma was monitored by ultrasound after two weeks. The rats were executed after one month. The testis and the accessory glands were removed and fixed, and the tissue structure and morphology were studied by immunofluorescence staining and HE staining. Results Eight rats were successfully injected. Apoptosis and necrosis in testicular convoluted seminiferous tubules were observed in different degrees. High expression of the reporter gene was observed in some tubules, and no obvious transfection was found in the epididymal epithelium. However, there was a high expression of the green fluorescent protein in seminal vesicle glandular epithelial cells. Conclusion Testis and epididymis are not suitable for lentivirus transfection in male animals for making transgenic animals, and the local transfection can easily affect sperm maturation and development. However, it is practical to establish transgenic animals by injecting lentivirus into the seminal vesicle.

Key words: Testicular efferent tubules, Lentivirus, Transgenic, Seminal vesicle, Epididymis

CLC Number: 

  • Q492.4
[1] 罗庆苗, 苗向阳, 张瑞杰, 等. 转基因动物新技术研究进展[J]. 遗传, 2011, 33(5): 449-458. LUO Qingmiao, MIAO Xiangyang, ZHANG Ruijie, et al. An update on the development of transgenic animal technology[J]. Hereditas, 2011, 33(5): 449-458.
[2] 郭宇航, 张守纯. 精子载体转基因技术的研究[J].畜禽业, 2008(8): 8-9.
[3] Russell LD, Ettlin RA, Hikim APS, et al. Histological and histopathological evaluation of the testis[J]. Int J Androl, 1993, 16(1): 83. doi:10.1111/j.1365-2605.1993.tb01156.x.
[4] Sun S, Wang YC, Guo K, et al. Finasteride inhibits microvascular density and VEGF expression in the seminal vesicle of rats[J]. Zhonghua Nan Ke Xue, 2018, 24(5): 387-392.
[5] Estemalik J, Demko C, Bissada NF, et al. Simultaneous detection of oral pathogens in subgingival plaque and prostatic fluid of men with periodontal and prostatic diseases[J]. J Periodontol, 2017, 88(9): 823-829.
[6] 曹碗君, 赵洁, 乔慧莲, 等. PTIP相关蛋白1在小鼠睾丸发育过程中的表达及定位[J]. 中国组织化学与细胞化学杂志, 2018, 27(1): 8-12. CAO Wanjun, ZHAO Jie, QIAO Huilian, et al. Expression and localization of PTIP associated protein 1 during mouse testis development[J]. Chinese Journal of Histochemistry and Cytochemistry, 2018, 27(1): 8-12.
[7] Jafarian A, Lakpour N, Sadeghi MR, et al. Transplantation of spermatogonial stem cells suspension into rete testis of azoospermia mouse model[J]. Urol J, 2018, 15(1): 40-47.
[8] Nistal M, González-Peramato P, Serrano Á. Acquired pathology of the rete testis[M] //Nistal M, González-Peramato P, Serrano Á. eds. Clues in the diagnosis of non-tumoral testicular pathology. Cham: Springer International Publishing, 2017: 279-285. doi:10.1007/978-3-319-49364-0_31.
[9] 廖波, 邓显忠, 代小思, 等. 成年SD大鼠睾丸网的分布[J]. 川北医学院学报, 2010, 25(3): 199-208. LIAO Bo, DENG Xianzhong, DAI Xiaosi, et al. Distribution of the rete testis in the adult SD rat[J]. Journal of North Sichuan Medical College, 2010, 25(3): 199-208.
[10] Martins-Santos E, Pimenta CG, Campos PRN, et al. Atrazine affects the morphophysiology, tissue homeostasis and aromatase expression in the efferent ductules of adult rats with mild alterations in the ventral prostate[J]. Chemosphere, 2018, 193: 958-967. doi:10.1016/j.chemosphere.2017.11.124.
[11] Michaelis M, Sobczak A, Weitzel JM. In vivo microinjection and electroporation of mouse testis[J]. J Vis Exp, 2014(90). doi:10.3791/51802.
[12] 王鹤, 曹文广. 小鼠睾丸输出管注射法建立转基因小鼠的试验研究[J]. 天津农业科学, 2013, 19(4): 34-37. WANG He, CAO Wenguang. Construction of transgenic mice by testicular efferent duct injection[J]. Tianjin Agricultural Sciences, 2013, 19(4): 34-37.
[13] 申复进, 张茨, 王将, 等. 输出小管-睾丸网注射提高小鼠精原干细胞移植效率[J]. 武汉大学学报(医学版), 2009, 30(4): 518-521. SHEN Fujin, ZHANG Ci, WANG Jiang, et al. Efferent duct-rete injection improves the efficiency for transplanting mouse spermatogonial stem cells[J]. Medical Journal Wuhan University, 2009, 30(4): 518-521.
[14] 周之畅, 李嘉诚, 胡宇峰, 等. 睾丸“门”的定位及临床解剖意义[J]. 现代实用医学, 2013, 25(10): 1097-1098. ZHOU Zhichang, LI Jiacheng, HU Yufeng, et al. The location of testis porta and its clinical significance in anatomy[J]. Modern Practical Medicine, 2013, 25(10): 1097-1098.
[15] 覃金洲. 雄性生殖细胞介导的转基因技术的研究[D]. 杨凌:西北农林科技大学,2016.
[16] Ning L, Meng J, Goossens E, et al. In search of an efficient injection technique for future clinical application of spermatogonial stem cell transplantation: infusion of contrast dyes in isolated cadaveric human testes[J]. Fertil Steril, 2012, 98(6): 1443-1448.e1. doi:10.1016/j.fertnstert.2012.08.023.
[17] Pease S. Infecting mouse embryos by coculturing with lentivirus[J]. Cold Spring Harb Protoc, 2018, 2018(3): pdb.prot093922. doi:10.1101/pdb.prot093922.
[18] 孟凡荣, 陈琛, 万海粟, 等. 慢病毒载体及其研究进展[J]. 中国肺癌杂志, 2014, 17(12): 870-876. MENG Fanrong, CHEN Chen, WAN Haisu, et al. Advances of lentiviral vectors[J]. Chinese Journal of Lung Cancer, 2014, 17(12): 870-876.
[19] Hashimoto M, Rockenstein E, Mante M, et al. An antiaggregation gene therapy strategy for Lewy body disease utilizing beta-synuclein lentivirus in a transgenic model[J]. Gene Ther, 2004, 11(23): 1713-1723.
[20] Kane NM, Nowrouzi A, Mukherjee S, et al. Lentivirus-mediated reprogramming of somatic cells in the absence of transgenic transcription factors[J]. Molecular Therapy, 2010, 18(12): 2139-2145.
[21] Bouchard M, Souabni A, Mandler M, et al. Nephric lineage specification by Pax2 and Pax8[J]. Genes Dev, 2002, 16(22): 2958-2970.
[22] 徐海生, 王长山, 李丽. PAX2基因在肾间质纤维化中的研究进展[J]. 齐齐哈尔医学院学报, 2017, 38(18): 2185-2188.
[23] Sinn PL, Arias AC, Brogden KA, et al. Lentivirus vector can be readministered to nasal epithelia without blocking immune responses[J]. J Virol, 2008, 82(21): 10684-10692.
[24] VandeWoude S, Hageman CL, Hoover EA. Domestic cats infected with lion or puma lentivirus develop anti-feline immunodeficiency virus immune responses[J]. J Acquir Immune Defic Syndr, 2003, 34(1): 20-31.
[1] XI Xianghong, SU Rong, JIA Shaotong, TANG Xiuying, WANG Lixin. Correlation between human epididymis protein 4 in seminal plasma and male infertility [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(5): 66-69.
[2] LI Maoyu, TANG Shi, DU Yifeng. Expression and significance of TRB3 in the brain of APP/PS1 transgenic mice [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(10): 1-5.
[3] XU Yi, ZHUANG Xianghua, LIU Yuantao, SUN Aili, LI Xiaobo, JIANG Dongqing, ZHANG Liang, XU Min, YAN Heng, CHEN Shihong. Over expression of lipin1 attenuates diabetic peripheral neuropathy in rats [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(9): 53-57.
[4] LUO Jiangang, SUN Tao, LIN Xiaowen, LI Yun, ZHAO Fei, MIAO Guishen, FU Zhijian. Lentivirus mediated RNA interference inhibits NF-κB p65 gene expression in spinal rat cord neurons [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(3): 23-26.
[5] ZHU Shou-rong1， ZHANG Rui2， CAO Yong-qian1， FENG Zhang1， WANG Yi-bing1. The effect of the survivin-siRNA targeting lentivirus vector on human malignant melanoma cell line A375 [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(5): 54-57.
[6] XU Yang-yang1,2， JIANG Zheng1,2， ZHOU Wei3， JIANG Yu-quan1, 2， LI Xin-gang1, 2. Construction of a lentiviral vector carrying shRNA against HIF-1α and generation of a GL261 cell line stably transfected with this vector [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(2): 12-16.
[7] SUN Ying1, DING Huan1, LI Xiao-qing2, LI Li1. Growth-inhibitory effect of over-expressed INPP4B in MDA-MB-231 cells via lentivirus transfection [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(12): 41-45.
[8] TIAN Bao-guo1, WANG Ping1, NIU Jian-yi2, WANG Zi-ping2, ZHAO Hai-qing2, BI Jian-zhong1. G-CSF and VEGF increase expressions of CD133 and synaptophysin in  the APP transgenic mouse model [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(1): 39-42.
[9] . Construction of lentiviral recombinants expressing shRNA targeting rPTTG 
gene and evaluation of knocking down efficiency mediated by lentivirus [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2009, 47(9): 76-80.
[10] CUI Li-ping,CUI Jing,SHI Chang-wen,SUN Qing. Package and expression of lentivirus encoding hTERT gene [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(2): 132-135.
[11] ZHOU Wei, FENG Jinbo, WANG Xuping, JIANG Hong, LIU Chunxi, WANG Rong, DING Shifang. A recombinant lentiviral vector carrying siRNA targeting the endothelial vascular growth factor a gene in mice [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(12): 1153-1157.
[12] WANG Jia-zeng,BAO Li-hua,SUN Jin-hao,LI Zhen-hua,ZHANG Jing. Survival and differentiation of GFP transgenic mice-derived neuroepithelial cells after being transplanted into rat brains [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2007, 45(7): 649-654.
[13] LIN A-li,SUN Qing. Package and expression of lentivirus encoding human GCSF genes [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2007, 45(6): 554-557.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd