In vivo gene transfer to mouse spermatogenic cells by deoxyribonucleic acid injection into seminiferous tubules and subsequent electroporation

Yukiko Yamazaki, Hirokazu Fujimoto, Hironori Ando, Takashi Ohyama, Yoshiko Hirota, Toshiaki Noce

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

An in vivo gene transfer technique for living mouse testes was used to develop a novel transient expression assay system for transcriptional regulatory elements of spermatogenic specific genes. The combination of DNA injection into seminiferous tubules and subsequent in vivo electroporation resulted in an efficient and convenient assay system for gene expression during spermatogenesis. The transfer of the firefly luciferase reporting gene driven by the Protamine-1 (Prm-1) enhancer region revealed a significant increase in the activity of the reporter enzyme. Histochemical studies of the transfer of the lacZ gene driven by the Prm-1 enhancer showed specific lacZ expression only in haploid spermatid cells in adult testes, corresponding with the expression pattern of endogenous Prm-1. We were able to detect long- lasting transgene expression in the transfected spermatogenic cells. A group of spermatogenic differentiating cells maintained the transfected lacZ expression after more than 2 mo of transfection, suggesting that spermatogenic stem cells and/or spermatogonia could also incorporate foreign DNA and that the transgene could be transmitted to the progenitor cells derived from a transfected proliferating germ cell.

Original languageEnglish
Pages (from-to)1439-1444
Number of pages6
JournalBiology of Reproduction
Volume59
Issue number6
Publication statusPublished - 1998 Dec
Externally publishedYes

Fingerprint

Seminiferous Tubules
Protamines
Electroporation
Transgenes
Injections
Testis
DNA
Stem Cells
Transcriptional Regulatory Elements
Genes
Firefly Luciferases
Gene Transfer Techniques
Spermatogonia
Lac Operon
Spermatids
Haploidy
Spermatogenesis
Germ Cells
Transfection
Gene Expression

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

In vivo gene transfer to mouse spermatogenic cells by deoxyribonucleic acid injection into seminiferous tubules and subsequent electroporation. / Yamazaki, Yukiko; Fujimoto, Hirokazu; Ando, Hironori; Ohyama, Takashi; Hirota, Yoshiko; Noce, Toshiaki.

In: Biology of Reproduction, Vol. 59, No. 6, 12.1998, p. 1439-1444.

Research output: Contribution to journalArticle

Yamazaki, Yukiko ; Fujimoto, Hirokazu ; Ando, Hironori ; Ohyama, Takashi ; Hirota, Yoshiko ; Noce, Toshiaki. / In vivo gene transfer to mouse spermatogenic cells by deoxyribonucleic acid injection into seminiferous tubules and subsequent electroporation. In: Biology of Reproduction. 1998 ; Vol. 59, No. 6. pp. 1439-1444.
@article{914224bb0c334d4e8bd6d2afba8a8df3,
title = "In vivo gene transfer to mouse spermatogenic cells by deoxyribonucleic acid injection into seminiferous tubules and subsequent electroporation",
abstract = "An in vivo gene transfer technique for living mouse testes was used to develop a novel transient expression assay system for transcriptional regulatory elements of spermatogenic specific genes. The combination of DNA injection into seminiferous tubules and subsequent in vivo electroporation resulted in an efficient and convenient assay system for gene expression during spermatogenesis. The transfer of the firefly luciferase reporting gene driven by the Protamine-1 (Prm-1) enhancer region revealed a significant increase in the activity of the reporter enzyme. Histochemical studies of the transfer of the lacZ gene driven by the Prm-1 enhancer showed specific lacZ expression only in haploid spermatid cells in adult testes, corresponding with the expression pattern of endogenous Prm-1. We were able to detect long- lasting transgene expression in the transfected spermatogenic cells. A group of spermatogenic differentiating cells maintained the transfected lacZ expression after more than 2 mo of transfection, suggesting that spermatogenic stem cells and/or spermatogonia could also incorporate foreign DNA and that the transgene could be transmitted to the progenitor cells derived from a transfected proliferating germ cell.",
author = "Yukiko Yamazaki and Hirokazu Fujimoto and Hironori Ando and Takashi Ohyama and Yoshiko Hirota and Toshiaki Noce",
year = "1998",
month = "12",
language = "English",
volume = "59",
pages = "1439--1444",
journal = "Biology of Reproduction",
issn = "0006-3363",
publisher = "Society for the Study of Reproduction",
number = "6",

}

TY - JOUR

T1 - In vivo gene transfer to mouse spermatogenic cells by deoxyribonucleic acid injection into seminiferous tubules and subsequent electroporation

AU - Yamazaki, Yukiko

AU - Fujimoto, Hirokazu

AU - Ando, Hironori

AU - Ohyama, Takashi

AU - Hirota, Yoshiko

AU - Noce, Toshiaki

PY - 1998/12

Y1 - 1998/12

N2 - An in vivo gene transfer technique for living mouse testes was used to develop a novel transient expression assay system for transcriptional regulatory elements of spermatogenic specific genes. The combination of DNA injection into seminiferous tubules and subsequent in vivo electroporation resulted in an efficient and convenient assay system for gene expression during spermatogenesis. The transfer of the firefly luciferase reporting gene driven by the Protamine-1 (Prm-1) enhancer region revealed a significant increase in the activity of the reporter enzyme. Histochemical studies of the transfer of the lacZ gene driven by the Prm-1 enhancer showed specific lacZ expression only in haploid spermatid cells in adult testes, corresponding with the expression pattern of endogenous Prm-1. We were able to detect long- lasting transgene expression in the transfected spermatogenic cells. A group of spermatogenic differentiating cells maintained the transfected lacZ expression after more than 2 mo of transfection, suggesting that spermatogenic stem cells and/or spermatogonia could also incorporate foreign DNA and that the transgene could be transmitted to the progenitor cells derived from a transfected proliferating germ cell.

AB - An in vivo gene transfer technique for living mouse testes was used to develop a novel transient expression assay system for transcriptional regulatory elements of spermatogenic specific genes. The combination of DNA injection into seminiferous tubules and subsequent in vivo electroporation resulted in an efficient and convenient assay system for gene expression during spermatogenesis. The transfer of the firefly luciferase reporting gene driven by the Protamine-1 (Prm-1) enhancer region revealed a significant increase in the activity of the reporter enzyme. Histochemical studies of the transfer of the lacZ gene driven by the Prm-1 enhancer showed specific lacZ expression only in haploid spermatid cells in adult testes, corresponding with the expression pattern of endogenous Prm-1. We were able to detect long- lasting transgene expression in the transfected spermatogenic cells. A group of spermatogenic differentiating cells maintained the transfected lacZ expression after more than 2 mo of transfection, suggesting that spermatogenic stem cells and/or spermatogonia could also incorporate foreign DNA and that the transgene could be transmitted to the progenitor cells derived from a transfected proliferating germ cell.

UR - http://www.scopus.com/inward/record.url?scp=0031796476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031796476&partnerID=8YFLogxK

M3 - Article

C2 - 9828190

AN - SCOPUS:0031796476

VL - 59

SP - 1439

EP - 1444

JO - Biology of Reproduction

JF - Biology of Reproduction

SN - 0006-3363

IS - 6

ER -