Male and female germline specific expression of an EGFP reporter gene in a unique strain of transgenic rats

Cronkhite, Jennifer T.; Norlander, Carola; Furth, Jenny K.; Levan, Göran; Garbers, David L.; Hammer, Robert E.

doi:10.1016/j.ydbio.2005.05.015

Cited by 36 publications

(37 citation statements)

References 33 publications

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“…Moreover, all postimplantation fetuses and newborns obtained from homozygous mothers were greenfluorescent. The same observations were reported for other transgenic strains of rats (Cronkhite et al 2005) and mice (Okabe et al 1997;Devgan et al 2004). …”

Section: Discussionsupporting

confidence: 85%

“…Cronkhite et al generated GFP transgenic rats under the control of the mouse Rosa26 promoter and detected fluorescence already in late 2-cell stage embryos recovered from WT females mated with homozygous transgenic males (Cronkhite et al 2005). In contrast, in a transgenic mouse line with a GFP gene under the control of the CAGGS promoter, the green fluorescence became visible from the 4-cell stage on (Okabe et al 1997).…”

Section: Discussionmentioning

confidence: 99%

“…Although transgenic rats expressing GFP genes in various tissues and organs have been generated (Hakamata et al 2001;Fujioka et al 2003;Takeuchi et al 2003;Inoue et al 2005;Francis et al 2007), only one study shows GFP expression in preimplantation rat embryos (Cronkhite et al 2005). However, there is no information about the reproductive performance and the ability of GFP transgenic rat embryos to develop in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

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Generation and characterization of a GFP transgenic rat line for embryological research

et al. 2008

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Model organisms expressing fluorescent proteins are important tools for research. The present study was performed to generate and characterize a new line of green fluorescent protein (GFP) transgenic rats for use as a model in experimental embryological research. We injected a GFP expression vector into 135 zygotes of the Sprague-Dawley (SD) rat strain. Embryo transfer of 103 surviving embryos resulted in the production of 35 offspring (33.9%) and two of them were transgenic (5.7%). Two transgenic rat lines that ubiquitously express GFP under the control of the cytomegalovirus-enhancer/beta-actin (CAGGS) promoter were generated by breeding. We studied the main embryological parameters of one these GFP transgenic lines. Homozygous GFP-transgenic females have the same ovulation and superovulation rates as wild type (WT) females. Transgenic embryos reached blastocyst stage in vitro and developed in vivo after embryo transfer without decrease in their developmental ability compared to the control group. The genotype of the parents determined the onset of GFP expression in preimplantation embryos. When the GFP gene is derived from the transgenic female parent, fluorescence was detected in oocytes and in embryos of all further stages of development. When the GFP gene is inherited by the transgenic male parent, GFP was only expressed from the blastocyst stage on. GFP-transgenic rats represent a valuable tool to mark embryos for many embryological studies such as transgenesis, gene expression patterns during early development, embryo aggregation for analysis of the distribution of cells in chimeric embryos and nuclear transfer to confirm the origin of the cloned offspring.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Generation and characterization of a GFP transgenic rat line for embryological research

et al. 2008

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“…The rat is a model of choice in several experimental settings (Aitman et al 2008) and recent advances in genetic engineering have resulted in the development of transgenic rats expressing reporter proteins such as b-galactosidase (Menoret et al 2002;Takahashi et al 2003) or enhanced-green fluorescent protein (eGFP) (Hakamata et al 2001;Ito et al 2001;Sawamoto et al 2001;Lois et al 2002;van den Brandt et al 2004;Cronkhite et al 2005;Inoue et al 2005;Itakura et al 2007;Michalkiewicz et al 2007). EGFP, a fluorescent protein derived from the jellyfish Aequorea Victoria (Prasher et al 1992), is an ideal marker for labelling cells, since its expression is stable in mammalian cells and its visualization in situ is easy, quantitative and non-invasive (Chalfie et al 1994).…”

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confidence: 99%

New lines of GFP transgenic rats relevant for regenerative medicine and gene therapy

et al. 2010

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Adoptive cell transfer studies in regenerative research and identification of genetically modified cells after gene therapy in vivo require unequivocally identifying and tracking the donor cells in the host tissues, ideally over several days or for up to several months. The use of reporter genes allows identifying the transferred cells but unfortunately most are immunogenic to wild-type hosts and thus trigger rejection in few days. The availability of transgenic animals from the same strain that would express either high levels of the transgene to identify the cells or low levels but that would be tolerant to the transgene would allow performing long-term analysis of labelled cells. Herein, using lentiviral vectors we develop two new lines of GFP-expressing transgenic rats displaying different levels and patterns of GFP-expression. The "high-expresser" line (GFP(high)) displayed high expression in most tissues, including adult neurons and neural precursors, mesenchymal stem cells and in all leukocytes subtypes analysed, including myeloid and plasmacytoid dendritic cells, cells that have not or only poorly characterized in previous GFP-transgenic rats. These GFP(high)-transgenic rats could be useful for transplantation and immunological studies using GFP-positive cells/tissue. The "low-expresser" line expressed very low levels of GFP only in the liver and in less than 5% of lymphoid cells. We demonstrate these animals did not develop detectable humoral and cellular immune responses against both transferred GFP-positive splenocytes and lentivirus-mediated GFP gene transfer. Thus, these GFP-transgenic rats represent useful tools for regenerative medicine and gene therapy.

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“…Rats in the SD-Tg(ROSA-EGFP)2-4Reh line were produced by pronuclear injection, and exhibit germ cell-specific expression of GFP (48). This allows germ cells (GFP ϩ ) to be clearly identified in cultures of testis cells prepared from this transgenic rat line (10,26).…”

Section: Methodsmentioning

confidence: 99%

Identification of Neuregulin as a Factor Required for Formation of Aligned Spermatogonia

Hamra

Chapman

Nguyen

et al. 2007

Journal of Biological Chemistry

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In the absence of somatic cells, medium conditioned by the SNL fibroblast line (SNL-CM) is able to stimulate primary cultures of rat type-A single spermatogonia to develop into chains of aligned spermatogonia at the 8-, 16-, and 32-cell stages. By comparison, medium conditioned by an MSC-1 Sertoli cell line is ineffective. Glial cell line-derived neurotrophic factor (GDNF)-like molecules were identified in SNL-CM and recombinant forms of GDNF, neurturin, and artemin were shown to stimulate formation of aligned spermatogonia, but principally to only the 4-and 8-cell stages. Because SNL-CM and GDNFlike molecules stimulated the formation of spermatogonial chain length differently, we purified components of SNL-CM to identify the additional contributing factor(s). A fraction was isolated that was dependent on GDNF, but required for effective formation of 16-and 32-cell chain lengths. Sequence analysis identified the factor as mouse neuregulin-1. At picomolar concentrations, recombinant neuregulin-1 in combination with GDNF effectively stimulated formation of aligned spermatogonia up to the 32-cell stage. Neuregulin in the absence of GDNF was relatively ineffective. Soluble receptors for neuregulins blocked the effects of GDNF and SNL-CM, suggesting that both neuregulin and GDNF are required for effective formation of long spermatogonial chains. Addition of neuregulin-1 to cultures on MSC-1 feeder layers resulted in spermatogonial behavior similar to that seen on feeder layers of SNL fibroblasts. In fact, SNL cells were found to express 100-fold higher levels of neuregulin-1 transcripts than MSC-1 cells. Thus, we identify neuregulin as a factor required for spermatogonial amplification and differentiation in culture.

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Male and female germline specific expression of an EGFP reporter gene in a unique strain of transgenic rats

Cited by 36 publications

References 33 publications

Generation and characterization of a GFP transgenic rat line for embryological research

Generation and characterization of a GFP transgenic rat line for embryological research

New lines of GFP transgenic rats relevant for regenerative medicine and gene therapy

Identification of Neuregulin as a Factor Required for Formation of Aligned Spermatogonia

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