본 연구는 이종간 생식세포 이식을 이용한 야생조류를 생산한 최초의 보고이다. 7일령 배자로부터 회수된 꿩 생식선 원시생식세포는 2.5일령 닭 배자의 혈관에 이식하였다. 생산된 카이메라에서 정상적인 꿩 유래의 생식세포가 spermatogenesis에 관여함을 꿩 정액 생산을 통해 확인하였고, 수컷 생식선 카이메라와 암컷 꿩의 번식을 통해 10마리의 꿩 자손을 생산하였다. 따라서 본 연구는 이종간 생식세포의 이식을 통해 야생조류의 생산을 가능하게 하였고, 이 기술은 멸종위기 조류의 복원에 적용될 수 있을 것이다.
Here, we describe the production of transgenic quail via a germline transmission system using postmigratory gonadal primordial germ cells (gPGCs). gPGCs retrieved from the embryonic gonads of 5-day-old birds were transduced with a lentiviral vector and subsequently transferred into recipient embryos. Testcross and genetic analyses revealed that among three germline chimeric G0 quail, one male produced transgenic offspring; of 310 hatchlings from the transgenic germline chimera, 24 were identified as donor-derived offspring, and 6 were transgenic (6/310, 1.9%). Conventional transgenesis using stage X blastodermal embryos was also conducted, but the efficiency of transgenesis was similar between the two systems (<1.6 vs. 1.9% for the conventional and gPGC-mediated systems, respectively). However, substantial advantages can be gained from gPGC-mediated method in that it enables an induced germline modification, whereas direct retroviral transfer to stage X embryos causes mosaic integration. The use of gonadal PGCs for transgenesis may lead to the production of bioreactors.
생식 세포는 한 세대의 유전 정보를 다음세대에 전달할 수 있는 유일한 세포로서 다양한 특성을 가지고 있다. 기능 유전체 연구를 통해서 새로운 유전자의 기능을 규명함으로써 그 유전자의 생물학적 의미와 상호 작용을 설명할 수 있다. 본 실험의 목적은 생식세포에서 특이적으로 발현하는 유전자를 발굴하여 그 유전자가 생식세포의 발달과 분화에 중요한 역할을 수행하는 것을 증명하는 것이다. 이에 본 실험에서는 real-time quantitative RT-PCR 기법을 이용하여 정소에서 특이적으로 발현하는 5개(AGE1, AGE2, AGE3, AGE4, AGE5)를 선발하였고, in situ hybridization 실험을 통하여 정소 조직 내에서의 발현 양상을 확인하였다. AGE1, AGE2 는 round spermatid에서 특이적으로 발현하고, AGE3, AGE4, AGE5는 spermatocyte에서 특이적으로 발현하는 것을 확인하였다. 본 실험을 통해 발굴한 유전자들은 닭의 생식선 발달에 중요한 기능을 할 것으로 예상되며, 앞으로 닭의 유전육종분야에 큰 도움을 줄 수 있을 것이다.
The aim of this study is to investigate the effects of ambient thermal environments on the development of swallowtail butterflies (Sericinus montela Gray). Developmental durations and survival rates of S. montela were examined at two crucial developmental stages, embryonic and larval development, at varying temperatures ranging from $15^{\circ}C$ to $35^{\circ}C$. As expected, our results indicated that increasing temperatures decreased the developmental duration and survival rate of the eggs. However, the larvae and pupae showed maximum survival rates at $20.0^{\circ}C$ and $25.0^{\circ}C$, and the represented durations were similar to those of the eggs. Larval development was stage-specific, revealing that the fourth and fifth instars at the later stages were more susceptible to temperature variation. When considering both parameters, the optimal development of S. montela occurred within the temperature range of $20.0-25.0^{\circ}C$. The lower threshold for the complete development of S. montela from eggs to eclosion of adults was calculated at $10.6^{\circ}C$ by linear regression analysis. The estimated value is similar to that of other endemic insects distributed in temperate climate zones, which indicates that S. montela belongs to a small group of swallowtails adjusted to low ambient temperatures. From the results, we predict that the full development of S. montela could be achieved within the temperature range of $17.5-30.0^{\circ}C$. Embryonic development ceased at both test temperature extremes, and no further larval development proceeded after the third instar at $35.0^{\circ}C$. These results suggest that embryogenesis can be significantly influenced by slight variations in the ambient thermal environment that fall below the optimal range.
Abstract Background Homozygous or compound heterozygous pathogenic variants in the thromboxane A synthase 1 ( TBXAS1 ) gene are associated with Ghosal hematodiaphyseal dysplasia (GHDD) which is characterized by defective hematopoiesis and increased bone density of long bones. Methods Patients 1 and 2 are identical twins, who presented with red blood cell transfusion‐dependent normocytic anemia and thrombocytopenia with bone marrow fibrosis and cortical bone thickening of long bones on plain radiograph. To clarify the etiology of their anemia and thrombocytopenia, whole blood was used for the DNA extraction and analyzed using next‐generation sequencing (NGS) on an in‐house bone marrow failure syndrome panel. Results The NGS results indicated that these two patients carried two heterozygous variants in TBXAS1 , exon7, c.583_584del, p.Ala195Leufs*12, and exon12, c.1420G>T, p.Gly474Trp, which were inherited from their mother and father, respectively. Patients 1 and 2 have been on chronic oral steroids with normalization of hemoglobin and platelet count after steroid initiation. Patient 3 is their sister who has normal blood counts but also has the same variants in TBXAS1 as her brothers. Radiographs showed cortical bone thickening and she has not required any treatment or transfusion. Conclusion We report three Caucasian siblings from non‐consanguineous parents with novel compound heterozygous variants of TBXAS1 presenting with the phenotypes of GHDD. These three cases illustrate the variable clinical expressivity of the GHDD from two‐compound heterozygous pathogenic variants of TBXAS1 .
Abstract Objective To measure the density of cellular phenotypes in canine caudal cruciate ligament (CaCL), cranial cruciate ligament (CrCL), medial collateral ligament (MCL), and long digital extensor tendon (LDET). Study design Ex‐vivo study. Methods Ten CaCL, CrCL, MCL, and LDET obtained from 1 stifle of 10 dogs with no gross pathology were analyzed histologically. The density of cells with 3 nuclear phenotypes (fusiform, ovoid, and spheroid) was determined within the core region of each specimen. Results Cells with fusiform nuclei were most dense in the MCL (median [range], 319 [118–538] cells/mm 2 ) and LDET (331 [61–463]), whereas cells with ovoid nuclei were most dense in the CaCL (276 [123–368]) and CrCL (212 [165–420]). The spheroid nuclear phenotype had the lowest density in all structures (31 [5–61] in CaCL, 54 [5–90] in CrCL, 2 [0–14] in MCL, and 5 [0–80] in LDET); however, the CrCL contained a denser population of spheroid cells compared with MCL and LDET ( P < .05). Total cell densities did not differ among the 4 structures ( P > .05). Conclusion Phenotype density varied within the ligaments and tendon tested here. The cell population of CaCL and CrCL differed from that of dense collagenous tissues such as MCL and LDET. Clinical significance The relatively higher density of spheroid phenotype in CrCL may reflect a distinctive native cellular population or a cellular transformation secondary to unique mechanical environment or hypoxia. This intrinsic cellular population may explain altered tissue properties prone to pathological rupture or poor healing potential of the canine CrCL.
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