Reconstitution of a chicken breed by inter se mating of germline chimeric birds

Abstract

Blastoderm cells from chicken embryos of a donor breed (Green-legged Partridgelike; GP) were transferred to embryos of a recipient breed (White Leghorn; WL) to form chimeric progeny that, after inter se mating, permitted successful reconstitution of the donor breed. Among 23 chimeric chicks hatched from WL embryos injected with GP cells, 20 (87%) were raised until maturity, and progeny were tested by mating with GP birds to determine the ability of blastodermal cells to form germline chimeras. Six of the tested birds (30%) produced recipient-derived and donor-derived offspring, indicating that they were germline chimeras. The mean percentages of donor-derived germ cells in these birds were 21.1 (17.6 to 50.0%) and 16.9 (5.3 to 23.1%) in males and females, respectively. Among 477 chicks, resulting from mating the germline chimeric male with four germline chimeric females, 10 chicks (2.1%) exhibited a GP phenotype, indicating that the original donor stock had been reconstituted. Only one germline chimeric hen produced GP offspring, but the expected and calculated percentages of GP offspring were similar (2.99 and 2.08, respectively). Two methods of DNA analyses (RFLP and PCR amplification of polymorphic microsatellite loci) of chimeras and their offspring indicated that through mating of a relatively small number of chimeras it is possible to reconstitute a highly diverse population.

Sex Steroids Level in Blood Plasma and Ovarian Follicles of the Chimeric Chicken

“…Chimeric chickens were bred in general by two different procedures: (i) blastodermal cells (BCs) containing primordial germ cells (PGCs) or their precursors, isolated from stage X (Eyal‐Giladi and Kochav, 1976) chicken embryo, injected into subgerminal cavity of recipient embryo (Marzullo, 1970); (ii) primordial germ cells isolated from germinal crescent, blood or gonads of donor embryos and transferred directly into bloodstream of the recipient embryos (Yasuda et al., 1992). In our laboratory, the production of chicken somatic and germline chimeras has been conducted (Bednarczyk et al., 2002, 2003) by the transfer of donor BCs into windowed eggs, according to the method proposed by Petitte et al. (1990), with some modifications (Bednarczyk et al., 2000).…”

“…Blastodermal cells are pluripotent and when transferred from donor to recipient embryos can contribute to different embryonic cell types including the germline. Chimerism is demonstrated routinely using feather (Petitte et al., 1990) and shell colouring (Czekalski, 2004), analysis of sex chromosomes from a sex‐mismatched donor (Simkiss et al., 1996; Kagami et al., 2002) or analysis of polymorphic DNA sequences (Bednarczyk et al., 2002; Naito et al., 2004) by using a marker. However, only few studies have attempted to study the effect of donor cells on physiology, development and production of post‐hatch chickens.…”

A Review of Strategies for Producing Chimeric Birds

“…A series of intraspecies chimeras and donor cell-derived offspring have been reported in previous studies using blastodermal cell transfer (Carsience et al, 1993;Kagami et al, 1997;Kino et al, 1997;Speksnijder and Ivarie, 2000;Naito et al, 2001). Based on intraspecies technology, intergenus and interorder germline chimeras have been obtained between Japanese quail and White Leghorn (WL) (Naito et al, 1991), Green-legged Partridgelike and WL (Bednarczyk et al, 2002), Maya duck and WL (Li et al, 2002), and Japanese quail and Peking duck as shown in Figure 1A, B (Gao et al, 2011). Intergenus and interorder germline chimeras and progenies were successfully produced by Green-legged Partridgelike-WL (Bednarczyk et al, 2002) and Maya duck-WL chimeras as shown in Figure 1C, D (Li et al, 2002) using blastodermal cell injection, but the efficiency of progeny production was low.…”

“…Based on intraspecies technology, intergenus and interorder germline chimeras have been obtained between Japanese quail and White Leghorn (WL) (Naito et al, 1991), Green-legged Partridgelike and WL (Bednarczyk et al, 2002), Maya duck and WL (Li et al, 2002), and Japanese quail and Peking duck as shown in Figure 1A, B (Gao et al, 2011). Intergenus and interorder germline chimeras and progenies were successfully produced by Green-legged Partridgelike-WL (Bednarczyk et al, 2002) and Maya duck-WL chimeras as shown in Figure 1C, D (Li et al, 2002) using blastodermal cell injection, but the efficiency of progeny production was low. In our latest experiments, we produced chimeras in different classes (Peking duck and Chinese soft-shelled turtle).…”

Secondary sexual traits and semen characteristic of chicken germline chimeras

“…Germline chimeras are one of the best tools for studying bird embryo development (Tagami & Kagami, ), as well as to produce the transgenic birds (Ono et al., ; Tajima, Naito, Yasuda, & Kuwana, ) or to reconstitute endangered species (Bednarczyk et al., ). Many years of bird embryo research provided different methods of localizing primordial germ cells (PGCs), which are used to obtain germline chimeras—beginning with periodic acid‐Schiff (PAS) staining technique (Ginsburg & Eyal‐Giladi, ; Muniesa & Dominguez, ), to immunostaining (Karagenç, Cinnamon, Ginsburg, & Petitte, ).…”