The effect of divergent selection for uterine capacity on fetal and placental development at term in rabbits: maternal and embryonic genetic effects

The aim of this work was to study the effects of the genotype of the dam, the embryo, or their interactions on prenatal growth by performing double-reciprocal embryo transfers between two lines of rabbits divergently selected for uterine capacity. Females from high (n = 53) and low (n = 48) lines were slaughtered at 72 h of gestation, and recovered embryos were transferred to the oviducts of recipient does from the high (n = 23) and low (n = 19) lines. Each recipient doe received eight embryos from the high line into one oviduct and eight embryos from the low line into the other. Recipient does were slaughtered on d 28 of gestation. The percentages of live fetuses at 28 d of gestation were 89.2 and 74% for high and low recipient lines, respectively. Length and weight of the empty uterine horn and weight of the full uterine horn were not affected by either the recipient or by donor line. Fetal weight was affected by the recipient line but not by the donor line. Fetuses gestated in high recipient does were 7% heavier (P < 0.10) than those in the low recipient does. There was a donor and a donor x recipient interaction effect on fetal placental weight. Fetal placental weight was heavier (7%, P < 0.01) for embryos from the low line. Embryos from the high line gestated in low-line uteri showed a lower fetal placenta weight than did low-line embryos gestated in high-line uteri and low-line uteri (P < 0.05). Linear regression coefficients of fetal weight at term on fetal placental weights differed (P < 0.05) for the high and low donors (4.33 +/- 0.28 and 3.41 +/- 0.29 respectively). A significant effect of the donor genotype on individual placental length was observed (P < 0.05), which might have resulted from a smaller individual placental length of low-line embryos gestated high-line uteri (P < 0.10). Neither donor nor recipient lines affected maternal placental weight or available space for fetuses. Fetuses and their fetal placentae were heavier when receiving more than four blood vessels than when receiving less than three blood vessels (13 and 17% respectively, P < 0.05). Neither recipient nor donor genotype affected the number of blood vessels arriving at each live fetus. Thus, fetal weight depends on the maternal genotype, whereas fetal placental weight depends on the embryo genotype in these two lines of rabbits divergently selected for uterine capacity.     

Induction of T cell anergy by the treatment with IL-10-treated dendritic cells

In order to apply for reducing graft versus host disease in allogeneic stem cell transplantation, the study concerning the induction of specific T cell anergy was designed. Normal allogeneic lymphocytes, which were co-cultured with IL-10-treated immature dendritic cells in the first mixed leukocyte culture (MLC), were cultured with mature dendritic cells of the same origin as IL-10-treated immature dendritic cells in the second MLC. By co-culturing with IL-10-treated immature dendritic cells, the response of normal lymphocytes to mature dendritic cells cultured from the same individual as that of IL-10-treated dendritic cells was markedly reduced, compared with the lymphocytes cultured with non-treated dendritic cells or IL-10-treated dendritic cells from a third party individual. The present study demonstrated that antigen specific T cell anergy was generated by priming allogeneic lymphocytes with IL-10-treated immature dendritic cells. These data suggested the applicability of IL-10-treated recipient dendritic cells for the induction of recipient cell-specific donor T cell anergy in donor graft.     

Mechanical transmission of porcine reproductive and respiratory syndrome virus by mosquitoes, Aedes vexans (Meigen)

The objective of this study was to determine whether porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted to naïve pigs by mosquitoes following feeding on infected pigs. During each of 4 replicates, mosquito-to-pig contact took place on days 5, 6, and 7 after PRRSV infection of the donor pig. A total of 300 mosquitoes [Aedes vexans (Meigen)] were allowed to feed on each viremic donor pig, housed in an isolation room. After 30 to 60 s, feeding was interrupted, and the mosquitoes were manually transferred in small plastic vials and allowed to feed to repletion on a naïve recipient pig housed in another isolation room. Prior to contact with the recipient pig, the mosquitoes were transferred to clean vials. Swabs were collected from the exterior surface of all vials, pooled, and tested for PRRSV. Separate personnel handled the donor pig, the recipient pig, and the vial-transfer procedure. Transmission of PRRSV from the donor to the recipient pig occurred in 2 out of 4 replicates. The PRRSV isolated from the infected recipient pigs was nucleic-acid-sequenced and found to be 100% homologous with the virus used to infect the donor pigs. Homogenates of mosquito tissues collected in all replicates were positive by either polymerase chain reaction or swine bioassay. All control pigs remained PRRSV negative, and PRRSV was not detected on the surface of the vials. This study indicates that mosquitoes (A. vexans) can serve as mechanical vectors of PRRSV.     

Manipulation of fish germ cell: visualization, cryopreservation and transplantation

Germ-cell transplantation has many applications in biology and animal husbandry, including investigating the complex processes of germ-cell development and differentiation, producing transgenic animals by genetically modifying germline cells, and creating broodstock systems in which a target species can be produced from a surrogate parent. The germ-cell transplantation technique was initially established in chickens using primordial germ cells (PGCs), and was subsequently extended to mice using spermatogonial stem cells. Recently, we developed the first germ-cell transplantation system in lower vertebrates using fish PGCs and spermatogonia. During mammalian germ-cell transplantation, donor spermatogonial stem cells are introduced into the seminiferous tubules of the recipient testes. By contrast, in the fish germ-cell transplantation system, donor cells are microinjected into the peritoneal cavities of newly hatched embryos; this allows the donor germ cells to migrate towards, and subsequently colonize, the recipient genital ridges. The recipient embryos have immature immune systems, so the donor germ cells can survive and even differentiate into mature gametes in their allogeneic gonads, ultimately leading to the production of normal offspring. In addition, implanted spermatogonia can successfully differentiate into sperm and eggs, respectively, in male and female recipients. The results of transplantation studies in fish are improving our understanding of the development of germ-cell systems during vertebrate evolution.     

Effect of the strain combination of the donor and recipient on the production efficiency of W‐bearing sperm in mixed‐sex germline chimeric chickens

Effect of the strain combination of the donor and recipient on production efficiency of W‐bearing sperm in mixed‐sex chimeric testes was analyzed. The combinations of the donors and recipients were White Leghorn (WL) and Rhode Island Red (RIR), and vice versa. Generated mixed‐sex chimeras that had the male phenotype at sexual maturity were classified into four groups: (1) a female WL donor and a male RIR recipient; (2) a male WL donor and a female RIR recipient; (3) a female RIR donor and a male WL recipient; (4) a male RIR donor and a female WL recipient. The mean number of W‐bearing sperm detected by in situ hybridization among 10 000 sperm observed were 147, 165, 30 and 45 in groups 1, 2, 3 and 4, respectively. The numbers in groups 1 and 2 were both significantly higher than those of groups 3 and 4 (P < 0.05). The combination of a WL donor and a RIR recipient produced W‐bearing sperm more efficiently than the reverse combination.     

Extracellular matrix in epitheliochorial,endotheliochorial and haemochorial placentation and its potential application for regenerative medicine

A placenta is defined as structural approximation of maternal and foetal tissues to perform physiological exchange. Associated processes of differentiation and the establishment of its cells take place within the extracellular matrix (ECM) that provides a rich environment of collagens, fibronectins, cytokines and other components. Placental ECM is promising for tissue regeneration purposes, because it has immune tolerance capacities that may cause only minimal rejections of transplants with immunological differences between donor and recipient. However, specific characteristics of ECM during evolution of the structurally very diverse mammalian placenta are not yet revealed. We here address the major aspects of placental types, that is non‐invasive (epitheliochorial), medium (endotheliochorial)‐to‐high (haemochorial) invasive nature of the interhemal barrier between the foetal and maternal blood system as well as their main components of ECM with special reference to species that are commonly used as animal models for human placentation and in the potential applications for regenerative medicine.     

异种动物核移植研究进展

异种核移植作为新型发展起来的动物繁殖技术 ,在畜牧生产、医学、生物学领域愈来愈表现出其巨大的价值。人们以前已经进行了一系列的探索 ,如在供体细胞方面 ,人们尝试使用了包括肝脏细胞、乳腺细胞在内的各种类型的细胞 ,并采取各种措施处理供体细胞 ;而对于受体细胞多采用卵泡 -卵丘复合体进行体外培养或通过超排获得 ;比较各种融合方法 ,目前多采用电融合方法 ,也较化学融合、病毒介导融合等方法简单 ;至于重构胚的培养则多采用培养液体外培养的方法 ,此方法操作简单 ,也比较容易观察记录实验过程。同时也指出异种核移植仍存在如克隆的效率很低等一系列急待解决的问题     

Early Kinetics of Infectious Hematopoietic Necrosis Virus (IHNV) Infection in Rainbow Trout

Abstract

A series of experiments was carried out with infectious hematopoietic necrosis virus (IHNV; 193-110 isolate) in rainbow trout Oncorhynchus mykiss (weight, ～1.2 g) to determine the duration of the patent period and the timing of onset of the infectious periods. We first attempted to transmit IHNV to recipient fish from infected rainbow trout 2–3 d after they had been exposed. No infection transfer occurred despite high titers (10^4.79 to 10^4.91 plaque-forming units 5–8 d postexposure (dpe). To determine the number of secondary cases produced by one infectious individual, we exposed approximately 50 rainbow trout (weight, ～1.5 g) in each of seven replicate tanks to a donor fish that had been infected with virus by bath exposure 3 d earlier. The prevalence of infection in recipient fish rose from 0.84% at 2 dpe to 7.9% at 6 dpe. Maximum incidence (22 cases) occurred between 2 and 4 dpe. No disease-specific mortalities occurred in recipient fish during the experiment. The titer of virus in both recipient and donor fish increased from 2 to 4 dpe. There was a positive correlation between the level of infection among donors and prevalence values among recipient fish (r ² = 0.60). The level of challenge by one infectious fish under the conditions provided was enough for infection transfer from sick cohabitant to susceptible fish but was not enough for initiation of a full-scale epizootic among recipients.     

The effect of divergent selection for uterine capacity on prenatal survival in rabbits: maternal and embryonic genetic effects

The aim of this work was to determine whether prenatal survival depends on the genotype of the mother or of the embryo and to identify the critical periods for prenatal mortality in two lines of rabbits divergently selected by high (H) and low (L) uterine capacity. Does from H (n = 124) and L (n = 115) lines were slaughtered at 72 h of gestation. Embryos recovered at 72 h of gestation were transferred to the oviducts of recipient does from the H (n = 23) and L (n = 19) lines. Each recipient does received eight embryos from the H line into one oviduct and eight embryos from the L line into the other. Recipient does were slaughtered on d 28 of gestation. No differences were found between lines in the embryo recovery either in ovulation rate (OR) or in fertilization rate of ova recovered. Recovery rate was higher for the H line (0.80 vs. 0.72, P < 0.01). The number of embryos recovered, fitting ovulation rate as a covariate, was also higher for the H line (9.74 vs. 8.78, P < 0.05). The H line showed a more advanced embryonic stage of development, having a higher percentage of blastocysts (PB) and a lower percentage of compact morulae (PCM) (38% vs. 20%, P < 0.001 for PB, and 51% vs. 64%, P < 0.01 for PCM). The percentage of early morulae was low and similar in both lines. Neither donor nor recipient lines affected embryonic survival from 72 h to 7 d of gestation. Fetal survival was affected by the recipient line (P < 0.05). An interaction between donor and recipient was found. Embryos from the H donor line had a better fetal survival rate than embryos from the L donor line (P < 0.05) in H recipient females. Within L recipient females, embryos from H and L donor lines showed similar fetal survival. Fetal survival was divided into early (from d 7 to 17 of gestation) and late (from d 17 to 28 of gestation). The high recipient line showed a higher early fetal survival than the L recipient line (P < 0.05). The same effect was observed for late fetal survival, but the difference between H and L recipient lines was lower (P < 0.10). Thus, fetal survival depends mainly on the maternal genotype, and the embryo genotype only affects fetal survival when embryo transfer is performed to a favorable maternal environment. Selection for uterine capacity in rabbits leads to modification of early embryonic survival and of early and late fetal survival, but differences are higher for early than for late fetal survival.     

Transcervical embryo transfer in performance mares

Pregnancy was established by transcervical transfer of embryos from performance mares into recipient mares. Estrus was synchronized between donor (n = 17) and recipient (n = 43) mares. After a greater than or equal to 25-mm follicle was detected, donor mares were bred artificially daily until ovulation. Day of ovulation was recorded. Uterine flushes (n = 111) were performed on donor mares 7 days after ovulation, and recovered embryos were transferred transcervically to recipient mares within 2 hours. Embryos were recovered from 40.5% of uterine flushes. Of transferred single embryos, 65.7% resulted in pregnancy, detectable by ultrasonographic examination 23 days after transfer. Only 35.3% of twin embryos resulted in pregnancy. Results over a 4-year period were as follows: uteri were flushed on 14, 44, 31 and 22 occasions, and 8, 21, 15, and 11 embryos were recovered (1 embryo was not transferred), with 6, 11, 4, and 6 resulting in 30-day pregnancy in years 1 to 4, respectively.     

绵羊胚胎移植生殖内分泌调控及其效果研究

初产供体母羊（14月龄）德国美利奴15只、波德代15只、无角陶赛特18只分别用孕酮(CIDR)+促性腺激素(PMSG)+促卵泡素(FSH)进行处理,受体小尾寒羊母羊（2胎）124只用CIDR+PMSG进行处理,处理期测定供、受体外周血液中生殖激素孕酮（p4）、雌二醇（E2）、促卵泡素（FSH）和促黄体素（LH）的动态变化。结果显示：3个供体品种和受体各自发情同期化水平和效果达到非常理想的状态,但供、受体发情同步差相距1 d。黄体数德国美利奴（7.71）显著高于陶赛特（5.19）（P<0.05）;冲卵数德国美利奴（5.50）显著高于陶赛特（3.44）（P<0.05）;可用胚胎率德国美利奴、波德代、陶赛特分别为70.13%、55.74%、78.18%,陶赛特显著高于波德代（P<0.05）;德国美利奴、波德代、陶赛特胚胎移植受胎率分别为55.26%、54.17%、42.31%,德国美利奴显著高于陶赛特（P<0.05）。处理期供体和受体生殖激素动态变化的幅度大,第13天供、受体血清中p4达到峰值,小尾寒羊分别高于德国美利奴（5.37 ng/mL）、波德代（6.14 ng/mL）和陶赛特（5.37 ng/mL）;供体血清中E2于第14天开始发情达到峰值,小尾寒羊于第15天开始发情达到峰值;供体血清中FSH于第13天达到峰值,小尾寒羊第14天达到峰值;供体血清中LH于第14天开始发情达到峰值,小尾寒羊血清中LH于第15天开始发情达到峰值。     

Transplantation of bovine germinal cells into mouse testes

To develop techniques for spermatogonial transplantation in bulls, it is essential to have an effective bioassay procedure to evaluate the transplantation efficiency of spermatogonial stem cell collection, purification, and culture techniques. The objective of the present study was to develop a mouse bioassay model to evaluate transplantation efficiency of fresh and cultured bovine germ cells. Bull calves of four ages (1, 2, 3, and 4 mo) were used as a source of donor testes cells. Two calves were used for each age point, one calf was experimentally made cryptorchidistic at 1 wk of age and the other left normal. A STO (mouse fibroblast) feeder cell line was used to culture bovine testes cells for 2 wk preceding transfer into recipient testes. Immunodeficient nude mice (nu/nu) in which endogenous spermatogenesis had been abolished by busulfan treatment served as recipient animals for transplantation. Donor bovine germ cells were microinjected into mouse seminiferous tubules. Mouse testes were analyzed 2 wk after transplant with the use of a bovine-specific antibody and whole-mount immunohistochemistry for the presence of bovine donor germ cells. Bovine testis cells were present in all recipient mouse testes analyzed. Fresh bovine testes cells were observed as colonies of round cells within mouse seminiferous tubules, indicating spermatogonial expansion and colonization; however, cultured bovine testes cells appeared as fibrous tissue and not as spermatogenic colonies. The average number of colonies resulting from donor cryptorchid testes was not different (P > 0.05) from noncryptorchid, 56+/-4 and 78+/-7, respectively. Fresh donor cells from calves older than 1 mo gave rise to a greater average number of colonies within recipient testes (P <0.05) (1 mo, 33+/-4; 2 mo, 70+/-8; 3 mo, 63+/-6; 4 mo, 87+/-9). Fresh bovine germ cells are capable of colonization in the busulfan-treated nude mouse testis, making it a suitable model for evaluation and development of spermatogonial transplant techniques in bulls.     

T-cell tolerance to the developing equine conceptus

One of the most intriguing and dramatic examples of immunological tolerance is displayed by the mammalian foetal-placental unit, which thrives as a semi-allograft in the mother's uterus during pregnancy. The success of the so-called foetal allograft stands in stark contrast to the failure of most tissue and organ grafts to survive without genetic matching of donor and recipient or drastic immunosuppression of the recipient's immune system. Experiments conducted over the past 60 years have revealed multiple mechanisms that enable the conceptus to avoid immunological detection or destruction. Many of these mechanisms are directed towards evading immune-mediated damage by maternal T lymphocytes, and they can be grouped into three classes: (i) downregulation of major histocompatibility complex (MHC) gene expression in placental trophoblast cells; (ii) local and systemic alterations in maternal immune reactivity; and (iii) innate defence mechanisms of the trophoblast cells that comprise the barrier between foetal and maternal tissues. The redundancy in these protective mechanisms helps ensure the transmission of life from generation to generation and provides a rich field of study of ways in which functional immunological tolerance can be manifest. The variation in placental forms and function among mammalian species present opportunities to discover and understand novel tolerogenic mechanisms that may have broad application in biology, medicine and animal husbandry. This review focuses on the evidence obtained from studies of pregnancy in the mare that support the case for selective T-cell tolerance to the mammalian conceptus.     

核移植前激活受体卵子对牛核移植卵体外发育的影响

本研究探讨了核移植前对受体卵子进行激活、细胞融合开始时间及供体受精卵细胞周期调节对核移植卵体外发育的影响。其结果显示核移植前对受体卵子激活组的细胞融合率与对照组没有差异 ,但重组胚胎的卵裂率、8～ 16细胞期胚胎及囊胚的发育率比对照组明显提高 ;核移植前激活的受体卵子分别在卵子体外成熟开始的第30h和 4 5h与供体细胞进行细胞融合 ,结果 ,30h组的细胞融合率和卵裂率与 4 5h组没有差异 ,但发育到 8～ 16细胞期及囊胚的发育率均比 4 5h的高 ;将供体受精卵用诺考达唑 (Nocodazole)处理后 ,进行核移植的结果 ,处理组的细胞融合率、卵裂率、发育到 8～ 16细胞期和囊胚的发育率与对照组无差异     

Multiple embryo-transplant offspring produced from quartering a bovine embryo at the morula stage

A non-surgical embryo collection was completed on a day 7 superovulated Chianina donor cow. Because all but two of the ova from the collection were unfertilised and a surplus of potential recipients was available, one embryo (an excellent quality late morula) was dissected into four equal portioned 'quarter' embryos using a simplified micromanipulation procedure. Each quarter embryo was then placed in a 0.25 ml French straw and non-surgically transplanted to four different crossbred beef recipient females. The remaining embryo was similarly transplanted to a herd mate recipient as an intact embryo. One recipient returned to oestrus, one recipient had an extended post transfer cycle and the two remaining recipients produced a live quarter embryo transplant calf each within 24 hours of the other. The intact embryo placed in a herd mate recipient did not produce a transplant calf. To the authors' knowledge, these transplant offspring are the first live births reported from a non-surgically collected later-stage bovine morula (day 7), which had been dissected into quarters and then individually transplanted non-surgically to recipient females. The procedure was relatively simple to perform and was completed in less than one hour.     

影响牛胚胎移植效果的主要因素分析

胚胎移植技术自创立以来,经过不断发展和完善,已成为现代畜牧业尤其是高价值低繁殖力畜种繁育中不可或缺的技术手段.但由于胚胎移植是一个系统工程,其移植效果受供受体的选择、超排效果、胚胎质量、供受体的同期性、操作技术、营养及饲养管理水平等诸多因素的影响,移植效果也参差不齐.本文就影响牛胚胎移植效果的主要因素进行简要分析,以期...     

Canine Haematopoietic Chimerism Analyses by Semiquantitative Fluorescence Detection of Variable Number of Tandem Repeat Polymorphism

Canine models are successfully applied to the study of haematopoietic stem cell transplantation (HSCT). Monitoring of haematopoietic donor/recipient chimerism is of major significance in detecting and quantifying engraftment or graft rejection of the donor-derived haematopoietic cells after transplantation. Radioactive analyses of polymorphic microsatellite markers are commonly used for chimerism analyses. We describe an improved, non-isotopic method that is based on the analysis of microsatellite markers in donor and recipient cells using capillary electrophoresis and fluorescence detection. Artificial mixtures of donor and recipient DNA that were generated from peripheral blood mononuclear cells from dog leukocyte antigen-identical siblings were used to analyse the sensitivity of the assay. DNA from dogs that had received HSCT were also analysed in order to demonstrate the feasibility of the method in vivo. For chimerism analyses, six different microsatellite loci were systematically amplified using fluorescent PCR primer. The fluorescent polymerase chain reaction products were separated by capillary electrophoresis using POP4 on a 310 ABI Prism Genetic Analyzer. After electrophoresis, fluorescence signals were automatically sized and quantified using GeneScan software. The method described provides an accurate assessment of haematopoietic chimerism in the canine model with significantly reduced hands-on time compared to conventional gel electrophoresis.     

Reproductive Parameters of Mangalarga Marchador Mares in a Commercial Embryo Transfer Programme

The objective of this study is to evaluate the reproductive efficiency in donors and recipient Mangalarga Marchador mares in commercial programmes of embryo transfer (ET) and the effects of some reproductive characteristics and ET methodology on conception rates in the recipient mares. A total of 1140 flushing procedures were performed and 830 embryos (72.8%) were recovered. There were no differences between the rates of embryonic recovery in the different breeding seasons (p > 0.05) and 92.8% of the recovered embryos were 8–9 days old. There was no difference in the embryonic recovery regarding the collection order from the first to the ninth embryo collection along the breeding season, as well as among mares inseminated during the foal heat or subsequent cycles (p > 0.05). Pregnancy rates observed in the total period of all reproductive seasons at 15, 30, 45 and 60 days of pregnancy were 73.4, 69.9, 66.7 and 64.5%, respectively. Differences in pregnancy rate and early embryonic loss rates were not observed between embryos transferred immediately after collection (66.8% and 13.5%) and embryos transported at room temperature for periods of <1 h (62.9% and 14.4%; p > 0.05). Pregnancy rates were higher when the interval between ovulations of donor and recipient mares remained between ?3 and ?2 days (p < 0.05), and the lowest rates were observed for intervals of ?6 days (p < 0.05) with intermediary values for intervals of ?1, 0 and +1 (p > 0.05). Embryonic loss rates, however, did not differ between intervals of ovulation’s synchronism between donor and recipient mares (p > 0.05). This flexibilization in the ovulatory synchronism between donor and recipient mares optimizes the use of recipient mares, thus reducing costs and facilitating management of horse breeding farms.     

Developmental capacity of reconstituted mouse embryos: influences of nucleus and cytoplasm sources.

This study was undertaken to examine the developmental capacity of reconstituted mouse embryos, and the influences of nucleus and cytoplasm on the development of these embryos following reciprocal pronuclear transplantation between in vitro 2-cell blocked and nonblocked embryos. Karyoplast containing pronuclei was transferred into the perivitelline space of the enucleated zygote and fused to cytoplasm with electrofusion. Maximum fusion rate was obtained when a field strength of 1.5 kV/cm was used. The fusion rates were high (86.2 +/- 3.2 to 90.6 +/- 2.0%) regardless of the strains of donor nucleus and recipient cytoplasm. Developmental rates of reconstituted embryos to the blastocyst stage, which were similar to that of the F1 (C57BL/6J x CBA) control were high when F1 embryos were used as the cytoplasm recipients (88.8 +/- 1.5 and 91.9 +/- 2.0%). When ICR embryos were used as the recipient cytoplasm, developmental rates were significantly reduced (71.5 +/- 2.9 and 54.1 +/- 3.2%), and affected by the source of nucleus. There were no significant differences in the cell number of embryos that developed to blastocysts and in the developmental rates to live young among the embryos reconstituted with different nuclei and cytoplasm, and the ICR control. The results of this study show that the development of reconstituted embryos is hardly affected by nuclear transplantation and electrofusion procedures. It is indicated that the recipient cytoplasm, rather than the donor nucleus, has the greater influence on the in vitro development of the reconstituted embryos to the blastocyst stage.     

Effect of soft X-ray irradiation on the migratory ability of primordial germ cells in chickens

1. The present study was conducted to elucidate the effect of soft X-ray irradiation on the migratory ability of primordial germ cells (PGCs) to the germinal ridges of chicken embryos. 2. PGCs (Barred Plymouth Rock, BPR) were isolated from embryonic blood and irradiated with soft X-rays for 1-10 min. Then, the PGCs were transfected in vitro with GFP gene by lipofection. The manipulated PGCs were transferred to recipient embryos (White Leghorn, WL) and migration to the germinal ridges was analysed by examining GFP gene expression in the gonads of recipient embryos under UV light at x40 magnifications. The expression of GFP gene was detected in all the gonads of recipient embryos examined up to 10.5 d of culture. 3. Migration of PGCs irradiated with soft X-rays to the germinal ridges was also confirmed by detecting a single nucleotide polymorphism in the D-loop region of the mitochondrial DNA of BPR and WL chickens. Freshly collected PGCs (BPR) were transferred to the bloodstream of recipient embryos (WL). The fate of the transferred donor PGCs was traced by detecting the single nucleotide polymorphism in the D-loop region of the mitochondrial DNA in BPR and WL used in this study. Transferred donor PGC-derived cells were detected in all the gonads of 17-d cultured embryos by PCR. 4. The results suggest that PGCs irradiated with soft X-rays still retain the ability to migrate to the germinal ridges of recipient embryos.