In Vivo Genetic Manipulation of Spermatogonial Stem Cells and Their Microenvironment by Adeno-Associated Viruses.
In Vivo Genetic Manipulation of Spermatogonial Stem Cells and Their Microenvironment by Adeno-Associated Viruses.
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通过腺体相关病毒对精子干细胞及其微环境的体内遗传操纵。
DOI：
10.1016/j.stemcr.2018.03.005
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发表时间：
2018-05-08
影响因子：
5.9
通讯作者：
Shinohara T
中科院分区：
文献类型：
--
作者：
Watanabe S;Kanatsu-Shinohara M;Ogonuki N;Matoba S;Ogura A;Shinohara T
腺相关病毒（AAV）可穿透血脑屏障，但尚不清楚 AAV 是否可穿透其他紧密连接。睾丸的基因操作受到曲细精管基底膜和血睾屏障（BTB）的阻碍，血睾屏障在支持细胞之间形成，并将曲细精管分为基底室和腔内室。在这里，我们展示了通过 AAV1/9 对精原干细胞 (SSC) 及其微环境进行体内遗传操作。显微注射到生精小管中的 AAV1/9 穿透基底膜和 BTB，从而不仅转导支持细胞和 SSC，还转导管周细胞和 Leydig 细胞。此外，当具有缺陷支持细胞的先天性不育Kit1S1/Kit1S1-d小鼠睾丸接受表达Kit1的AAV时，精子发生再生并产生后代。没有一个后代含有 AAV 基因组。因此，AAV1/9 可以通过穿透 BTB 和基底膜进行有效的种系和生态位操作，为生殖缺陷基因疗法的开发提供了一种有前途的策略。 AAV 穿透血睾屏障并感染 SSC 及其环境 AAV 还穿透生精小管的基底膜 AAV 可以挽救先天性不育 KitlSl/KitlSl-d 小鼠的精子发生 显微授精后后代不携带转基因 在这篇文章中，Shinohara 及其同事展示了腺相关病毒 (AAV) 穿透血睾屏障和睾丸基底膜的可行性小鼠睾丸的生精小管。 AAV 不仅感染生殖细胞，还感染其微环境。该技术用于挽救支持细胞缺陷的不孕小鼠模型。
Adeno-associated virus (AAV) penetrates the blood-brain barrier, but it is unknown whether AAV penetrates other tight junctions. Genetic manipulation of testis has been hampered by the basement membrane of seminiferous tubules and the blood-testis barrier (BTB), which forms between Sertoli cells and divides the tubules into basal and adluminal compartments. Here, we demonstrate in vivo genetic manipulation of spermatogonial stem cells (SSCs) and their microenvironment via AAV1/9. AAV1/9 microinjected into the seminiferous tubules penetrated both the basement membrane and BTB, thereby transducing not only Sertoli cells and SSCs but also peritubular cells and Leydig cells. Moreover, when congenitally infertile KitlSl/KitlSl-d mouse testes with defective Sertoli cells received Kitl-expressing AAVs, spermatogenesis regenerated and offspring were produced. None of the offspring contained the AAV genome. Thus, AAV1/9 allows efficient germline and niche manipulation by penetrating the BTB and basement membrane, providing a promising strategy for the development of gene therapies for reproductive defects. AAVs penetrate the blood-testis barrier and infect SSCs and their environment AAVs also penetrate the basement membrane of the seminiferous tubules AAVs could rescue spermatogenesis in congenitally infertile KitlSl/KitlSl-d mice Offspring did not carry transgenes after microinsemination In this article, Shinohara and colleagues show the feasibility of adeno-associated virus (AAV) to penetrate the blood-testis barrier and the basal membrane of the seminiferous tubules of mouse testes. AAVs infected not only germ cells but also their microenvironment. The technique was used to rescue a mouse model of infertility with Sertoli cell defect.
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影响因子：
3.6
作者：
Blanchard, KT;Boekelheide, K
通讯作者：
Boekelheide, K
影响因子：
5.1
作者：
Dalkara, D.;Byrne, L. C.;Flannery, J. G.
通讯作者：
Flannery, J. G.
DOI：
10.1073/pnas.022646399
发表时间：
2002-02-05
影响因子：
11.1
作者：
Kanatsu-Shinohara, M;Ogura, A;Shinohara, T
通讯作者：
Shinohara, T
DOI：
10.1073/pnas.0506084102
发表时间：
2005-11-15
影响因子：
11.1
作者：
Meng, J;Holdcraft, RW;Braun, RE
通讯作者：
Braun, RE
影响因子：
12.4
作者：
Dalkara, Deniz;Kolstad, Kathleen D.;Flannery, John G.
通讯作者：
Flannery, John G.