Adenovirus-mediated gene transfer to rat testis in vivo
Adenovirus-mediated gene transfer to rat testis in vivo
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DOI：
10.1095/biolreprod56.2.495
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发表时间：
1997-02-01
影响因子：
3.6
通讯作者：
Boekelheide, K
中科院分区：
文献类型：
--
作者：
Blanchard, KT;Boekelheide, K
为了研究体内成年大鼠睾丸中的转基因表达，使用携带具有核定位信号的lacZ转基因的腺病毒载体作为标记物。腺病毒载体首先在体外大鼠支持细胞-生殖细胞共培养物上进行了测试;它有效地介导了支持细胞中的转基因表达，但不是生殖细胞。然后通过睾丸内注射将该载体递送至成年大鼠睾丸的间质区室，导致Leydig细胞表达转基因。或者，通过睾丸网注射将载体递送至小管内隔室导致生精上皮的支持细胞和附睾的主细胞表达转基因。在体内，每种细胞类型在感染后2天表达转基因，并且表达持续至少10天;然而，稍后的时间点与转基因表达的丧失和局灶性间质炎症相关。本研究证明了腺病毒介导基因转移到特定睾丸细胞的能力，为研究体内特定基因对精子发生的短期影响提供了有力的工具。
To study transgene expression in the adult rat testis in vivo, an adenovirus vector carrying a lacZ transgene with a nuclear localization signal was used as a marker. The adenovirus vector was first tested on rat Sertoli cell-germ cell cocultures in vitro; it efficiently mediated transgene expression in Sertoli cells but not germ cells. This vector was then delivered to the interstitial compartment of adult rat testes by intratesticular injection, resulting in Leydig cells expressing the transgene. Alternatively, delivering the vector to the intratubular compartment by rete testis injection resulted in expression of the transgene by Sertoli cells of the seminiferous epithelium and principal cells of the epididymis. In vivo, each cell type expressed the transgene by 2 days postinfection, and expression persisted for at least 10 days; however, later time points were associated with a loss of transgene expression and focal interstitial inflammation. This study documents the ability of adenovirus to mediate gene transfer to specific testicular cells, providing a powerful tool to study the short-term effects of specific genes on spermatogenesis in vivo.