Availability of subfertile transgenic rats expressing the c-myc gene as recipients for spermatogonial transplantation

Masumi Hirabayashi, Yusuke Yoshizawa, Megumi Kato, Takashi Tsuchiya, Shizuko Nagao, Shinichi Hochi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The spermatogonial transplantation system was applied to evaluate stem cell kinetics and niche quality and to produce gene-modified animals using the stem cells after homologous recombination-based selection. This study was designed to determine whether the transplanted spermatogonia were able to proliferate and differentiate in male rats expressing the c-myc transgene under control of the human metallothionein IIA promoter (MT-myc Tg rats). Donor testicular cells were prepared from heterozygous chicken β actin (CAG)/enhanced green fluorescent protein (EGFP)-transgenic rats (EGFP Tg rats) during the second week after birth and injected into the seminiferous tubules of the MT-myc Tg rats (line-A and -B; both subfertile) or rats pretreated with busulfan to remove endogenous spermatogonia. Three to four months after transplantation, cell colonies with EGFP fluorescence were detected in 36% (4/11), 40% (8/20), and 71% (5/7) of the transplanted testes in line-A MT-myc Tg rats, line-B MT-myc Tg rats, and busulfan-treated rats, respectively. No EGFP-positive colonies were detected when wild-type male rats were used as recipients (0/7; testis-basis). The histopathological and immunofluorescent examination of the serial sections from the transplanted testes showed normal spermatogenesis of the donor spermatogonia, but atrophy of the recipient seminiferous tubules. Microinsemination with round spermatids and mature spermatozoa derived from EGFP-positive testes in line-A rats resulted 26% (10/39 transferred) and 23% (11/48 transferred) full-term offspring, respectively. Thus, the MT-myc Tg male rats were suitable as potent recipients for spermatogonial transplantation without any chemical pretreatment to remove the endogenous spermatogonia.

Original languageEnglish
Pages (from-to)135-141
Number of pages7
JournalTransgenic Research
Volume18
Issue number1
DOIs
Publication statusPublished - 01-02-2009

Fingerprint

Transgenic Rats
myc Genes
Transplantation
genetically modified organisms
rats
Spermatogonia
spermatogonia
genes
green fluorescent protein
Testis
testes
busulfan
Busulfan
Seminiferous Tubules
seminiferous tubules
stem cells
Stem Cell Niche
Spermatids
Metallothionein
Homologous Recombination

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Genetics
  • Agronomy and Crop Science
  • Animal Science and Zoology

Cite this

Hirabayashi, Masumi ; Yoshizawa, Yusuke ; Kato, Megumi ; Tsuchiya, Takashi ; Nagao, Shizuko ; Hochi, Shinichi. / Availability of subfertile transgenic rats expressing the c-myc gene as recipients for spermatogonial transplantation. In: Transgenic Research. 2009 ; Vol. 18, No. 1. pp. 135-141.
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abstract = "The spermatogonial transplantation system was applied to evaluate stem cell kinetics and niche quality and to produce gene-modified animals using the stem cells after homologous recombination-based selection. This study was designed to determine whether the transplanted spermatogonia were able to proliferate and differentiate in male rats expressing the c-myc transgene under control of the human metallothionein IIA promoter (MT-myc Tg rats). Donor testicular cells were prepared from heterozygous chicken β actin (CAG)/enhanced green fluorescent protein (EGFP)-transgenic rats (EGFP Tg rats) during the second week after birth and injected into the seminiferous tubules of the MT-myc Tg rats (line-A and -B; both subfertile) or rats pretreated with busulfan to remove endogenous spermatogonia. Three to four months after transplantation, cell colonies with EGFP fluorescence were detected in 36{\%} (4/11), 40{\%} (8/20), and 71{\%} (5/7) of the transplanted testes in line-A MT-myc Tg rats, line-B MT-myc Tg rats, and busulfan-treated rats, respectively. No EGFP-positive colonies were detected when wild-type male rats were used as recipients (0/7; testis-basis). The histopathological and immunofluorescent examination of the serial sections from the transplanted testes showed normal spermatogenesis of the donor spermatogonia, but atrophy of the recipient seminiferous tubules. Microinsemination with round spermatids and mature spermatozoa derived from EGFP-positive testes in line-A rats resulted 26{\%} (10/39 transferred) and 23{\%} (11/48 transferred) full-term offspring, respectively. Thus, the MT-myc Tg male rats were suitable as potent recipients for spermatogonial transplantation without any chemical pretreatment to remove the endogenous spermatogonia.",
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Availability of subfertile transgenic rats expressing the c-myc gene as recipients for spermatogonial transplantation. / Hirabayashi, Masumi; Yoshizawa, Yusuke; Kato, Megumi; Tsuchiya, Takashi; Nagao, Shizuko; Hochi, Shinichi.

In: Transgenic Research, Vol. 18, No. 1, 01.02.2009, p. 135-141.

Research output: Contribution to journalArticle

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AU - Hirabayashi, Masumi

AU - Yoshizawa, Yusuke

AU - Kato, Megumi

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AU - Nagao, Shizuko

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