Effects of busulfan on murine spermatogenesis: cytotoxicity, sterility, sperm abnormalities, and dominant lethal mutations

Bucci, Luke R.; Meistrich, Marvin L.

doi:10.1016/0027-5107(87)90057-1

Cited by 229 publications

(178 citation statements)

References 21 publications

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“…Sertoli cells constitute only 3% of the wild-type testis cell population (31) and are more difficult to target in the wild-type testis than in the infertile testis. Because the number of Sertoli cells remains constant in the adult testis and is not affected by busulfan treatment (2,32), the elimination of germ cells increased the relative concentration of Sertoli cells in the testis cell population. Second, Sertoli cells are the only cells that would be directly exposed to injected adenovirus, because other types of cells are separated by the blood-testis barrier between Sertoli cells (2).…”

Section: Discussionmentioning

confidence: 99%

Adenovirus-mediated gene delivery and in vitro microinsemination produce offspring from infertile male mice

Kanatsu-Shinohara

Ogura²,

Ikegawa³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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Sertoli cells play a pivotal role in spermatogenesis through their interactions with germ cells. To set up a strategy for treating male infertility caused by Sertoli cell dysfunction, we developed a Sertoli cell gene transfer system by using an adenovirus vector, which maintained long-term transgene expression in the testes of infertile mice. Introduction of an adenovirus carrying the mouse Steel (Sl) gene into Sertoli cells restored partial spermatogenesis in infertile Steel͞Steel dickie (Sl͞Sl d ) mutant mouse testes. Although these males remained infertile, round spermatids and spermatozoa from the testes produced normal fertile offspring after intracytoplasmic injection into oocytes. None of the offspring showed evidence of germ line transmission of adenoviral DNA. Thus, we demonstrate a successful treatment for infertility by using a gene therapy vector. Therefore, adenovirus-mediated gene delivery into Sertoli cells not only provides an efficient and convenient means for studying germ cell-Sertoli cell interactions through manipulation of the germ cell microenvironment in vivo, but also is a useful method to treat male infertility resulting from a Sertoli cell defect.

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Section: Discussionmentioning

confidence: 99%

Adenovirus-mediated gene delivery and in vitro microinsemination produce offspring from infertile male mice

Kanatsu-Shinohara

Ogura²,

Ikegawa³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…The administration of the anticancer drug busulfan caused severe reduction in the number of spermatogenic cells within 8 weeks, but spermatogenesis recovered from the damage during the next 12 weeks (Figure 4A), as reported previously. 21 Annexin V or control reagents were microinjected into the seminiferous tubules 8 weeks after treatment with busulfan so that the reagents could affect spermatogenesis from the beginning of recovery, and the testes were analyzed for the occurrence of spermatogenesis at week 20 ( Figure 4B). We found that the histology of the testis sections prepared from annexin Vinjected mice was almost equal to that obtained at week 8. This indicates that spermatogenesis did not recover from the damage caused by busulfan when testes received microinjection of annexin V. Injection of either buffer alone or BSA did not influence the recovery of spermatogenesis.…”

Section: Inhibition Of Sperm Production In Annexin V-treated Micementioning

confidence: 99%

“…21 After 8 weeks, the mice were microinjected with annexin V, BSA or buffer alone. Epididymides were isolated, soaked in M2 medium (Sigma) supplemented with 0.5% BSA, and disrupted with forceps.…”

Section: Analysis Of Epididymal Spermmentioning

confidence: 99%

Inhibition of sperm production in mice by annexin V microinjected into seminiferous tubules: possible etiology of phagocytic clearance of apoptotic spermatogenic cells and male infertility

et al. 2002

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Many differentiating spermatogenic cells die by apoptosis during the process of mammalian spermatogenesis. However, very few apoptotic spermatogenic cells are detected by histological examination of the testis, probably due to the rapid elimination of dying cells by phagocytosis. Previous in vitro studies showed that Sertoli cells selectively phagocytose dying spermatogenic cells by recognizing the membrane phospholipid phosphatidylserine (PS), which is exposed to the surface of spermatogenic cells during apoptosis. We examined here whether PS-mediated phagocytosis of apoptotic spermatogenic cells occurs in vivo. For this purpose, the PS-binding protein annexin V was microinjected into the seminiferous tubules of normal live mice, and their testes were examined. The injection of annexin V caused no histological changes in the testis, but significantly increased the number of apoptotic spermatogenic cells as assessed by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay. The number of Sertoli cells did not change in the annexin V-injected testes, and annexin V itself did not induce apoptosis in primary cultured spermatogenic cells. These results indicate that annexin V inhibited the phagocytic clearance of apoptotic spermatogenic cells and suggest that PS-mediated phagocytosis of those cells occurs in vivo. Furthermore, the injection of annexin V into the seminiferous tubules brought about a significant reduction in the number of spermatogenic cells and epididymal sperm in anticancer drug-treated mice. This suggests that the elimination of apoptotic spermatogenic cells is required for the production of sperm.

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“…Recipient mice are treated with busulfan to destroy endogenous spermatogenesis (Bucci & Meistrich, 1987). This allows transplanted spermatogonia to reach the basement membrane of the seminiferous tubule (Brinster & Avarbock, 1994).…”

Section: Mice and Cell Collectionmentioning

confidence: 99%

Culture of mouse spermatogonial stem cells

et al. 1998

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Spermatogenesis occurs within the seminiferous tubules of mammals by a complex process that is highly organized, extremely efficient and very productive. At the foundation of this process is the spermatogonial stem cell that is capable of both self-renewal and production of progeny cells, which undergo differentiation over a period of weeks to months in order to generate mature spermatozoa. It had been thought that germ cells survive only a brief period in culture, generally less than a few weeks. However, an accurate assessment of the presence of spermatogonial stem cells in any cell population has only recently become possible with development of the spermatogonial transplantation technique. Using this technique, we have demonstrated that mouse spermatogonial stem cells can be maintained in culture for approximately 4 months and will generate spermatogenesis following transplantation to the seminiferous tubules of an appropriate recipient. Extensive areas of cultured donor cell-derived spermatogenesis are generated in the host, and production of mature spermatozoa occurs. Cultivation of the testis cells on STO feeders is beneficial to stem cell survival. These results provide the first step in establishing a system that will permit spermatogonial stem cells to be cultivated and their number increased in vitro to allow for genetic modification before transplantation to a recipient testis.

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Effects of busulfan on murine spermatogenesis: cytotoxicity, sterility, sperm abnormalities, and dominant lethal mutations

Cited by 229 publications

References 21 publications

Adenovirus-mediated gene delivery and in vitro microinsemination produce offspring from infertile male mice

Adenovirus-mediated gene delivery and in vitro microinsemination produce offspring from infertile male mice

Inhibition of sperm production in mice by annexin V microinjected into seminiferous tubules: possible etiology of phagocytic clearance of apoptotic spermatogenic cells and male infertility

Culture of mouse spermatogonial stem cells

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