The deleted in azoospermia like (DAZL) gene has been identified in many vertebrate species. DAZL shows high homology with deleted in azoospermia (DAZ) genes that identified only in humans, great apes and Old World monkeys, and boule homolog (BOLL) that identified in many vertebrate species. These genes encode RNA binding proteins (RBP), which regulate the post-transcriptional functions of several genes. In humans, DAZ copies are linked to Y chromosome, while DAZL and BOLL are linked to chromosomes 3 and 2, respectively. DAZ copies has been reported to express in prenatal and postnatal germ cells, particularly in the premeiotic spermatogonia. BOLL has been reported to express during the meiotic G2/M transition in germ cells. DAZL has been reported to express in all stages of germ cells. Compared to humans and mice, the detailed functionalities of DAZL is not clear in many vertebrate species. In our studies, we use chickens as an animal model to examine the expression profiling of DAZL gene in germ cells right from the early embryonic development to the adult. Also, we are studying the effects of small interfering RNA (siRNA) mediated knockdown of DAZL and Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/CRISPR associated protein 9 (CRISPR/Cas9) mediated knockout of DAZL in the chicken primordial germ cells (PGCs). In the chicken, DAZL is linked to chromosome 2 (2p1.3-p1.2), and encodes a 289 amino acids protein. By in situ hybridization, we detected a strong expression of DAZL in the germ plasm of chicken oocytes. Later, the expression of DAZL was strongly detected in all stages of intrauterine development and post-ovipositional development especially in the PGC specifying cells. Moreover, the expression of DAZL was strong and constant in the male and female germ cells until adult stage. The siRNA mediated knockdown of DAZL significantly reduced the PGCs proliferation and increased the apoptosis in vitro. We examined the knockout efficiency of DAZL using CRISPR/Cas9 technique in chicken DF1 fibroblast cell line, prior to test in the PGCs. The results of T7 endonuclease I (T7E1) assay and subsequent sequencing indicates clear mutations on the DAZL gene in DF1 cells, and the method could be applicable to cause mutations on the DAZL gene in PGCs. In conclusion, chicken DAZL express in all stages of germ cells as a germ line marker, and alteration in the gene expression causes germ cells impairment.

목 적 : Contrast enhancement MRA(이하 CE-MRA)를 검사할 때 가장 중요한 것은 조영제가 관심혈관을 지나가는 시기(조영제의 혈관도달/통과시간)를 포착하는 것이다. 대동맥과 같이 검사영역이 넓은 부위를 검사할 때는 검사영역을 여러 단계로 나눠서 검사해야 한다. 일반적으로 CE-MRA의 acquisition time은 조영제가 관심혈관을 통과하는 시간보다 길기 때문에 1단계 영역의 동맥상(arterial phase)에 비하여 2단계 영역의 동맥상이 늦어질 수 있다. 따라서 두 검사영역의 동맥상이 일치되도록 하기 위한 방안이 필요하다. 본 논문에서는 Multi-Steps CE-MRA 검사 시 동맥상의 차이를 최소화하고자 test bolus와 k-space time to center 개념을 접목한 방법을 사용하여 그 유용성을 평가하였다. 대상 및 방법 : 조영제를 사용하여 복부 MRI 검사를 하는 환자를 대상으로 Multi-steps CE-MRA 검사를 시행하였다. 대동맥궁부터 총장골동맥을 두 개의 영역으로 나눠서 MRA 검사를 조영 전/후에 각각 시행하였다. Test bolus scan을 시행하여 관심혈관의 조영제 통과시간을 측정하였다. 대조군 실험으로 상부영역과 하부영역의 time to center를 최소로 적용하여 CE-MRA를 실시하였다. 실험군 실험으로 1단계 영역은 k-space time to center를 최대로 설정하였고 2단계 영역은 최소로 적용하였다. 대조군과 실험군에서 얻은 영상으로부터 venous contamination 여부를 평가하였다. 또한 1단계 영역과 2단계 영역의 signal intensity를 측정하여 평가하였다. 결 과 : K-space timing control method를 사용하므로써 venous contamination이 더 적게 발생하였으며 검사영역 사이의 signal intensity도 더 균일하게 나타났다. 결 론 : K-space timing control method는 multi-steps CE-MRA에서 발생할 수 있는 venous contamination 발생을 해결할 수 있는 방법이며 이 방법을 활용하면 보다 편리하고 정확한 검사에 도움이 될 것이라고 사료된다.

In the present study, we have developed a high-frequency plant regeneration system for Italian ryegrass via callus culture using mature seeds as explants. Optimal embryogenic callus induction was found to occur in MS medium containing 5 ㎎ 1?¹ 2,4-D, 0.5 ㎎ 1?¹ BA, 500 ㎎ 1?¹ L-proline, 1 g 1?¹ casein hydrolysate, 30 g 1?¹ sucrose, 7 ㎎ 1?¹ AgNO₃, 2 ㎎ 1?¹ CuSO₄ and solidified with 3 g 1?¹ Gelrite. The highest regeneration rate was obtained in MS medium containing 1 ㎎ 1?¹ 2,4-D, 5 ㎎ 1?¹ BA, 500 ㎎ 1?¹ L-proline, 1 g 1?¹ casein hydrolysate, 1 ㎎ 1?¹ thiamine-HCl, 30 g 1?¹ sucrose, 7 ㎎ 1?¹ AgNO₃, 2 ㎎ 1?¹ CuSO₄ and solidified with 3 g 1?¹ Gelrite. By using the most effective treatment determined for each parameter, the highest rates of embryogenic callus formation (48.9%) and regeneration (47.6%) were obtained with the Hwasan 101 cultivar. The overall plant regeneration rates of the examined cultivars ranged from 7.5% to 23.2%. Thus, optimization of regeneration frequency using mature seeds as explant material may offer a simple and efficient protocol for Italian ryegrass that may improve molecular breeding of this species.