Hibbitt Olivia, Coward Kevin, Kubota Hiroki, Prathalingham Nilendran, Holt William, Kohri Kenjiro, Parrington John
Department of Pharmacology, University of Oxford, Oxford OX1 3QT, United Kingdom.
Biol Reprod. 2006 Jan;74(1):95-101. doi: 10.1095/biolreprod.105.042267. Epub 2005 Sep 14.
The study of gene function in testis and sperm has been greatly assisted by transgenic mouse models. Recently, an alternative way of expressing transgenes in mouse testis has been developed that uses electroporation to introduce transgenes into the male germ cells. This approach has been successfully used to transiently express reporter genes driven by constitutive and testis-specific promoters. It has been proposed as an alternative method for studying gene function in testis and sperm, and as a novel way to create transgenic animals. However, the low levels and transient nature of transgene expression that can be achieved using this technique have raised concerns about its practical usefulness. It has also not been demonstrated in mammals other than mice. In this study, we show for the first time that in vivo gene transfer using electroporation can be used to express a fluorescent transgene in the testis of a mammal other than mice, the Syrian golden hamster. Significantly, for the first time we demonstrate expression of a transgene in epididymal sperm using this approach. We show that expression of the transgene can be detected in sperm for as long as 60 days following gene transfer. Finally, we provide the first systematic demonstration that this technique does not lead to any significant long-term adverse effects on testicular integrity and sperm quality. This technique therefore offers a novel way to study gene function during fertilization in hamsters and may also have potential as a way of creating transgenic versions of this important model species.
转基因小鼠模型极大地推动了睾丸和精子中基因功能的研究。最近,一种在小鼠睾丸中表达转基因的替代方法被开发出来,该方法利用电穿孔将转基因导入雄性生殖细胞。这种方法已成功用于瞬时表达由组成型和睾丸特异性启动子驱动的报告基因。它被提议作为研究睾丸和精子中基因功能的替代方法,以及创建转基因动物的新途径。然而,使用该技术所能实现的转基因表达水平较低且具有瞬时性,这引发了人们对其实用性的担忧。此外,除小鼠外,该技术尚未在其他哺乳动物中得到验证。在本研究中,我们首次表明,利用电穿孔进行体内基因转移可用于在除小鼠外的另一种哺乳动物——叙利亚金仓鼠的睾丸中表达荧光转基因。重要的是,我们首次证明了使用这种方法可在附睾精子中表达转基因。我们发现,在基因转移后长达60天的时间里,均可在精子中检测到转基因的表达。最后,我们首次系统地证明了该技术不会对睾丸完整性和精子质量造成任何显著的长期不良影响。因此,这项技术为研究仓鼠受精过程中的基因功能提供了一种新方法,并且作为培育这种重要模式物种转基因版本的一种方式也可能具有潜力。