南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (07): 840-843.

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体内原位电穿孔致雄性SPFKM小鼠睾丸损伤的实验研究

袁进1, 安靓1, 顾为望2, 黄吴健1   

  1. 1. 南方医科大学组织胚胎学教研室, 广东, 广州, 510515;
    2. 南方医科大学实验动物中心, 广东, 广州, 510515
  • 出版日期:2005-07-20 发布日期:2005-07-20
  • 基金资助:
    收稿日期:2005-3-6。
    基金项目:广州市重点科技项目(JB02-2-024-01-2)
    作者简介:袁进(1969- ),男,在读硕士研究生,E-mail:yuanjin@fimmu.com,电话:020-61648204
    通讯作者:安靓,广州南方医科大学组织胚胎学教研室,E-mail:anjing@fimmu.edu.com, 电话: 020-61648205

Experimental study of in situ electroporation-induced testis injury in specific pathogen-free male Kunming mice

YUAN Jin1, AN Jing1, GU Wei-wang2, HUANG Wu-jian1   

  1. 1. 南方医科大学组织胚胎学教研室, 广东, 广州, 510515;
    2. 南方医科大学实验动物中心, 广东, 广州, 510515
  • Online:2005-07-20 Published:2005-07-20

摘要: 目的 利用电穿孔仪对成年雄性SPFKM小鼠睾丸进行电穿孔实验,以观察电穿孔对成年雄性SPFKM小鼠睾丸的损伤,为体内精子介导基因转移提供依据。方法 设置高压和低压两组不同的体内电穿孔参数,将SPFKM小鼠睾丸置于电穿孔仪上的钳型电极的两侧进行体内电穿孔,2周后处死动物,分离睾丸和附睾,将睾丸称重后用4%多聚甲醛固定,常规组织切片和HE染色,显微镜下观察;将附睾置于M16培养基中分离其中精子,稀释后置显微镜下观察。结果 (1)高压电穿孔2周后,附睾精子活力检查和睾丸组织切片镜下观察显示:高压电穿孔可造成睾丸的不可逆损伤;其睾丸质量也显著低于对照组睾丸质量(P<0.01)。(2)低压电穿孔2周后,附睾精子活力检查和睾丸组织切片镜下观察显示:低压电穿孔虽可造成睾丸一定的损伤,但这种损伤是可逆的,经过一段时间的恢复后雄鼠仍具有生殖能力。并且电穿孔后的睾丸质量与对照组睾丸质量不存在显著差异(P>0.05)。结论 合理地设计体内电穿孔参数,一方面不会对SPFKM雄鼠睾丸的生殖能力造成严重影响,另一方面它还可为外源基因导入到生殖细胞提供了一种可能。

Abstract: Objective To observe testis injuries induced by in situ electroporation in specific pathogen-free (SPF) adult male Kunming mice. Methods With two sets of parameters of high and low voltages,respectively,in situ electroporation of the testis was performed in vivo by fixing the testis and epididymis of the mice between a pair of rectangular tweezer-type electrodes.Two weeks after electroporation,the mice were killed and the testis and epididymis separated and fixed with 4% paraformaldehyde after recording the weight of the testis.Routine histological sections were prepared and observed under optical microscope after HE staining.The epididymis was transfered into M16 medium for spermatozoa separation,and after dilution of the spermatozoa suspension,the spermatozoa viability was observed under optical microscope. Results Two weeks after high-voltage electroporation,examination of spermatozoa viability and microscopy of the testis sections revealed irreversible testis injury,and the testis weight was significantly reduced in comparison with that of control mice (P<0.01).Low-voltage electroporation,in contrast,only caused reversible injuries of the testis,and the male mice retained their repro-ductive capacity after a certain length of recovery period.The testis weight after low-voltage electroporation showed no significant difference from that of the control mice (P>0.05). Conclusions Appropriate setting of the parameters for in vivo electroporation may avoid severe impact on the reproductive capacity of the testis in SPF male Kunming mice.This technique also provides a possibility for exogenous gene transfer into the reproductive cells.

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