Avian transgenesis: progress towards the promise.

Avian pluripotent stem cells.

The chick embryo is a classic model that has been used to gain insight into developmental processes and cell fate within the embryo for over a century. For the most part, investigators have implanted quail cells into a chicken embryo. A more powerful tool for developmental biology research than the quail:chick chimera system would be to have lines of transgenic chickens expressing reporter genes that are readily available to the research community. However, avian transgenic technology has been fraught with technical difficulties, and transgenic chickens expressing reporter genes have only recently been developed. The goal of this review is to report the technologies that have been used to generate transgenic chickens and to discuss the challenges in generating avian transgenics for developmental biology research.

Status of transgenic chicken models for developmental biology.

In order to provide an alternative to day-old chick culling in the layer hatcheries, a noninvasive method for egg sexing is required at an early stage of incubation before onset of embryo sensitivity. Fluorescence and Raman spectroscopy of blood offers the potential for precise and contactless in ovo sex determination of the domestic chicken (Gallus gallus f. dom.) eggs already during the fourth incubation day. However, such kind of optical spectroscopy requires a window in the egg shell, is thus invasive to the embryo and leads to decreased hatching rates. Here, we show that near infrared Raman and fluorescence spectroscopy can be performed on perfused extraembryonic vessels while leaving the inner egg shell membrane intact. Sparing the shell membrane makes the measurement minimally invasive, so that the sexing procedure does not affect hatching rates. We analyze the effect of the membrane above the vessels on fluorescence signal intensity and on Raman spectrum of blood, and propose a correction method to compensate for it. After compensation, we attain a correct sexing rate above 90% by applying supervised classification of spectra. Therefore, this approach offers the best premises towards practical deployment in the hatcheries.

/pdf/sexing-of-chicken-eggs-by-fluorescence-and-raman-107yt2h873.pdf

Sexing of chicken eggs by fluorescence and Raman spectroscopy through the shell membrane.

Consistent Production of Transgenic Chickens Using Replication-Deficient Retroviral Vectors and High-Throughput Screening Procedures

Stage X chick blastoderms following ovi- position were accessed via a small window in the egg. Windowing, however, substantially reduces the hatch- ability of eggs containing early embryos. For example, only 32 of 389 (8.2%) eggs hatched after standard win- dowing with or without irradiation or injection. Ex ovo culture systems can overcome this problem but are labor intensive. A modification of a standard windowing tech- nique has yielded an average hatch rate of 32% for 892 windowed eggs independent of incubator type, γ-irradia- tion, or injection of the embryo. This was a fourfold in- crease over a standard windowing method. Similar hatch rates were observed using fertile eggs from five chicken lines (Barred Plymouth Rock (BR), Athens-Canadian

Research Note A Modified Method of Shell Windowing for Producing Somatic or Germline Chimeras in Fertilized Chicken Eggs

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Research Note A Modified Method of Shell Windowing for Producing Somatic or Germline Chimeras in Fertilized Chicken Eggs