한국 재래 닭 고환 간질세포의 미세구조에 관한 연구

장병귀,태현진,최철환,박영재,양홍현,김남수,박상열,강형섭,박병용,이영훈,안동춘,김인식,Jang, B.G.,Tae, H.J.,Choi, C.H.,Park, Y.J.,Yang, H.H.,Kim, N.S.,Park, S.Y.,Kang, H.S.,Park, B.Y.,Lee, Y.H.,Ahn, D.C.,Kim, I.S. 한국가금학회 2006 韓國家禽學會誌 Vol.33 No.3

한국 재래 닭에서 부화 후부터 성숙에 이르는 시기까지 고환 발달동안 간질세포의 미세구조를 구명하기 위하여 부화 후 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 28, 32, 44, 52 및 64주령(n=13마리/일령)의 한국 재래 닭을 이용하여 이 연구를 수행하였다. 한국 재래닭의 고환은 2.5% glutaraldehyde를 이용하여 전신관류고정하고 조직 처리 과정을 거쳐 Epon-araldite에 포매하였다. 초박절편기를 사용하여 $1{\mu}m$로 절편한 다음 methylene blue로 염색하여 일반적인 조직의 변화상을 관찰하였고 간질세포의 미세구조는 초박 절편을 제작하여 투과전자현미경으로 관찰하였다. 간질세포내에서 테스토스테론과 연관성이 있는 미세구조인 사립체, 평활형질내세망 및 지방방울의 변화에 따라 간질세포의 발달 유형을 분류하면, 1) 간질세포가 치밀하게 배열되어 있고 세포 사이의 공간이 거의 없으며 핵이 간질세포 용적의 많은 부분을 차지하고 있는 시기로 핵은 전자밀도가 높은 유형과 희박한 유형으로 나눌 수 있었고, 또한 세포질 내에는 많은 양의 지방방울, 소량의 사립체 및 평활형질내세망을 갖고 있는 시기, 2) 세포질 내에 큰 폭으로 확장된 평활형질내세망, 풍부한 사립체 및 감소된 지방방울을 갖고 있는 시기로 나눌 수 있다. Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.

한국 재래 닭 부화 후 고환 발달에 관한 연구

장병귀,태현진,최철환,박영재,박병용,박상열,강형섭,김남수,이영훈,양홍현,안동춘,김인식,Jang, B.G.,Tae, H.J.,Choi, C.H.,Park, Y.J.,Park, B.Y.,Park, S.Y.,Kang, H.S.,Kim, N.S.,Lee, Y.H.,Yang, H.H.,Ahn, D.C.,Kim, I.S. 한국가금학회 2006 韓國家禽學會誌 Vol.33 No.3

이 연구는 한국 재래 닭에서 부화 후 고환 발달 과정을 명확하게 이해가기 위하여 부화 후 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 28, 32, 44, 52 및 64주령(n=13마리/일령)의 한국 재래닭을 이용하여 수행하였다. 한국 재래 닭의 고환은 2.5 % glutaraldehyde를 이용하여 전신 관류 고정하고 조직 처리 과정을 거쳐 Epon-araldite에 포매하였다. 초박절편기를 사용하여 $1{\mu}m$로 절편한 다음 methylene blue로 염색하여 일반적인 조직의 변화상과 형태 계측을 일반적인 방법에 따라서 정자 생성을 측정하여 다음과 같은 결과를 얻었다. 부화 후 1주령의 고환의 평균 무게는 0.015 g이었고 점진적으로 증가하여 21주령에는 3.93 g이고 21주령부터 64주령까지는 변화가 없었다. 곱슬정세관의 용적 치밀도는 1주령에 32.6%이었으나 점차적으로 증가하여 64주령에서는 92.89이었다. 1주령의 한국 재래 닭 고환 간질 조직은 고환 실질의 67.4%를 나타내었고 이러한 비율은 성장하는 동안에 점차적으로 감소하여 64주령에 7.11%를 나타내었다. 고환내 총 정자 생성은 18주령부터 28주령까지는 유의성있게 증가하였고 고환 1g당 정자 생성은 $18\sim28$주령까지는 유의성있게 증가하였고 $28\sim52$주령까지는 변화가 없었으나 64주령에 유의성 있게 감소하였다. 곱슬정세관의 평균 직경은 $1\sim21$주령까지 주령에 따라 점진적으로 증가하였고 곱슬정세관의 길이는 1주령에 0.34 m이었고 성장하면서 유의성 있게 증가하여 64주령에서는 72.2 m이었다. 곱슬정세관내 생식세포의 발달 단계는 다음과 같이 분류할 수 있다. 1) 정조세포($1\sim8$주령), 2) 정조세포, 정모세포($10\sim12$주령), 3) 정조세포, 정모세포, 원형의 정자세포($14\sim16$주령), 4) 정조세포, 정모세포, 정자세포 및 정자($18\sim64$주령). 이러한 결과를 종합하여 보면 한국 재래 닭에서 부화 후부터 성숙시기까지 고환 발달의 양상은 신생시기-성 성숙 이전기($1\sim12$주령), 성 성숙시기($14\sim18$주령) 및 성숙시기$(21\sim64)$로 나뉜다. Changes in the chicken testis from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The present study was to investigate in more detail the post-hatching development of testis in Korean native chickens. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. Sperm production was measured by routine technique. The average volume of a testis of 1 week old Korean native chickens was determined as 0.015 g and the parameter increased linearly from 1 week to 21 weeks days (28.9 g), and did not change from 21 weeks to 64 weeks. The volume density of the seminiferous tubules increased with age from 32.6% at week 1 to 92.89% at week 64. The volume density of the interstitium represents 67.4% of the testicular parenchyma at week 1. This proportion progressively diminished during development to reach a value of 7.11% at week 64. Total sperm production per testis increased significantly from 18 weeks to 28 weeks and remained unchanged. Sperm production per 1 g testis increased significantly from 18 weeks to 28 weeks, did not change significantly from 28 weeks to 52 weeks, and declined significantly at 64 weeks of age. The average diameter of the seminiferous tubules gradually increased with age from 1 week $(42.4{\mu}m)$ to 21 weeks $(412.8{\mu}m)$. The length of the seminiferous tubules was 0.34 m at 1 week, increased significantly in subsequent age groups and reached 72.2 m by weeks 64. The stage of germ cell development in seminiferous tubules was classified as 1) spermatogonia $(1\sim8\;weeks)$, 2) spermatogonia and spermatocytes $(10\sim12\;weeks)$, 3) spermatogonia, spermatocytes and round spermatids $(14\sim16\;weeks)$, and 4) speramatogonia, spermatocytes, spermatids and spermatozoa $(18\sim64\;weeks)$. These results clarified the pattern of changes in the testicular development in Korean native chickens from hatching to adulthood as 1) neonatal-prepubertal $(1\sim12\;weeks)$, 2) puberty$(14\sim18\;weeks)$, and adult$(21\sim64\;weeks)$.

Flavin mononucleotide (1,4-butanediamine) Pt(II) Complex와 Cisplatin의 세포주기에 대한 유세포 분석 및 ICR계 생쥐에서의 신장독성에 대한 생화학적 분석

권영이(Y. E. Kwon),황규자(K. J. Whang),김안근(A. K. Kim),김국환(K. H. Kim),김원규(W. K. Kim),안동춘(D. C. Ahn) 대한약학회 2000 약학회지 Vol.44 No.2

Flavin mononucleotide (1,4-butanediamine) Pt(II) complex (7FMN) was synthesized and screened anticancer activity [J.Pharm.Soc.Korea 43(6), 762-770 (1999)]. 7FMN have good water solubility and moderate anticancer activity. In this paper, cell-cycle specificity and nephrotoxicity were studied. Interaction of DNA with cisplatin and synthesized 7FMN was analyzed by flow cytometry, and showed G2 arrest in L1210 cell fine. It means that cell-cycle on L1210 was inhibit in S phase by cisplatin and 7FMN. In order to biochemically analyze nephrotoxicity of cisplatin and 7FMN, after injecting each agent intrapenritoneary, blood was exsangumated after 6 hours, 1 day, 3 days and 7 days, respectively. Then, serum was separated from the blood. The serum level of BUN, creatinine and uric acid in cisplatin and 7FMN administrated mice (25-35g, ICR strain, a dose each 8, 12 and 16 times of the IC50 value, cisplatin; 7 times) were determined by autochemistry analyzer. In cisplatin-administered mice group, BUN level was elevated than normal control group at 3rd day and repaired at 7th day. In 7FMN administrated group was not elevated. Crea@e and uric acid level were no difference with the normal control group. Therefore synthesized 7FMN is less toxic than cisplatin in nephrotoxicity.