Kojima Yoshiyuki, Hayashi Yutaro, Kurokawa Satoshi, Mizuno Kentaro, Sasaki Shoichi, Kohri Kenjiro
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Fertil Steril. 2008 May;89(5 Suppl):1448-54. doi: 10.1016/j.fertnstert.2007.04.062. Epub 2007 Jul 10.
To investigate the risk of germ-line transmission of vector sequences after in vivo adenovirus-mediated gene transfer to mouse testes and to discuss whether an adenovirus vector could be used in the future to treat male factor infertility.
Experimental animal study.
Laboratory research setting in the Department of Nephro-urology at Nagoya City University Graduate School of Medical Sciences in Japan.
ANIMAL(S): Eight-week-old B6C3F1 mice.
INTERVENTION(S): Adenovirus vector carrying a LacZ transgene as a marker was injected into the interstitial space (intratesticular injection) or seminiferous tubules (intratubular injection) of the mouse testis.
MAIN OUTCOME MEASURE(S): An assessment by polymerase chain reaction (PCR) and histological analyses of the proportion of adenovirus vectors administered into the testis that can infect epididymal sperm and transmit to fetuses derived from these males 3, 7, 14, 28, and 35 days after intratesticular or intratubular adenovirus injection.
RESULT(S): No PCR signal was identified in genomic DNA extracted from the epididymal sperm of all mice on each day after intratesticular or intratubular adenovirus injection. On reverse transcriptase (RT)-PCR analysis of mRNA isolated from fetuses derived from these males on each day after intratesticular or intratubular adenovirus injection, no fetuses had amplified products, although about 30% of the fetuses generated by microinjection into fertilized eggs had LacZ transcripts. On histochemical staining, no two-cell and 12.5 d.p.c. fetuses showed beta-gal activity. These sperm and fetus studies showed that adenovirus-mediated gene transfer to the testis does not cause infection of or transmission to the germ line or fetuses.
CONCLUSION(S): The risk of germ-line transmission after adenovirus-mediated gene transfer to the testis is extremely low, and this method can be exploited in the future for the treatment of male factor infertility.
研究体内腺病毒介导的基因转移至小鼠睾丸后载体序列发生种系传递的风险，并探讨未来腺病毒载体是否可用于治疗男性因素不育症。
实验动物研究。
日本名古屋市立大学医学研究生院肾泌尿学系的实验室研究环境。
8周龄的B6C3F1小鼠。
将携带LacZ转基因作为标记的腺病毒载体注射到小鼠睾丸的间质间隙（睾丸内注射）或生精小管（管内注射）。
通过聚合酶链反应（PCR）评估以及组织学分析，确定在睾丸内或管内注射腺病毒后3、7、14、28和35天，注入睾丸的腺病毒载体中可感染附睾精子并传递至这些雄性小鼠所产胎儿的比例。
在睾丸内或管内注射腺病毒后的每一天，从所有小鼠附睾精子中提取的基因组DNA均未检测到PCR信号。对睾丸内或管内注射腺病毒后每一天从这些雄性小鼠所产胎儿中分离的mRNA进行逆转录酶（RT）-PCR分析，尽管通过显微注射到受精卵中产生的胎儿约30%有LacZ转录本，但没有胎儿有扩增产物。组织化学染色显示，没有二细胞期和12.5天胚龄的胎儿显示β-半乳糖苷酶活性。这些精子和胎儿研究表明，腺病毒介导的基因转移至睾丸不会导致种系或胎儿感染或传递。
腺病毒介导的基因转移至睾丸后发生种系传递的风险极低，该方法未来可用于治疗男性因素不育症。
