Seminiferous tubule transfection in vitro to define post-meiotic gene regulation

Reproductive Biology and Endocrinology : RB&E
Sandra DannerRichard Ivell

Abstract

Post-meiotically expressed genes in the testis are essential for the proper progression of spermatogenesis, and yet, aside from the construction of individual transgenic mice using specific promoters to drive reporter plasmids, there are only very limited possibilities for relevant and quantitative analysis of gene promoters. This is due to the special nature of post-meiotic haploid cells, which to date are not represented in any appropriate cell-lines. This article reports the development of novel methodology using isolated and cultured rat seminiferous tubules in a multiwell format, into which promoter-reporter constructs can be introduced by a combination of microinjection and electroporation. Culture conditions were developed which allowed the continued incubation of isolated rat seminiferous tubules for up to 48 h without obvious cell death and loss of post-meiotic cells. Transfection of intact seminiferous tubules by microinjection and electroporation was optimized to achieve high expression efficiencies of control plasmids, using either fluorescent protein or luciferase as reporters, thereby allowing both morphological as well as quantitative assessment. Successful transfection was achieved into all cell types except for...Continue Reading

References

Apr 1, 1992·Molecular Reproduction and Development·K NayerniaW Engel
Aug 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·J J PeschonR D Palmiter
Jun 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·B P ZambrowiczR D Palmiter
Jul 15, 1999·Biochemical and Biophysical Research Communications·H S LiuN J Ke
Jan 5, 2000·The Journal of Experimental Zoology·Y YamazakiK Imoto
Sep 27, 2003·Developmental Biology·P Prabhakara ReddiKshitish K Acharya
Jan 8, 2004·Biological & Pharmaceutical Bulletin·Yoshinobu Nakanishi, Akiko Shiratsuchi
Jun 14, 2005·Developmental Biology·Masanobu ShojiNorio Nakatsuji

Citations

Jan 15, 2010·Nucleic Acids Research·Mirjana RajkovicJoachim M Weitzel
Nov 19, 2011·Morphologie : Bulletin De L'Association Des Anatomistes·C Ravel, S Jaillard
Sep 29, 2011·Molecular Reproduction and Development·Tushna DehnugaraM R Satyanarayana Rao
May 4, 2012·Spermatogenesis·Damien HunterRichard Ivell
Sep 28, 2018·Stem Cell Research & Therapy·Keykavos GholamiMehdi Abbasi

Related Concepts

Venus protein, Bacteria
Bacterial Proteins
Cell Survival
Luciferases
Luminescent Proteins
Microinjections
Early Promoters, Genetic
Seminiferous Tubule Structure
Structure of Sertoli Cell
Transfection

Related Feeds

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.