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생체밖에서 미국흰불나방 지방세포에 의한 저장단백질의 흡수와 축적에 관하여
이봉희,김관선,문명진,김우갑,Lee, Bong-Hee,Kim, Kwan-Seon,Moon, Myung-Jin,Kim, Woo-Kap 한국현미경학회 1988 Applied microscopy Vol.18 No.2
This study was carried out to examine in vitro first whether the storage proteins, which the fat bodies of last larvae from Hyphantria cunea secrete into haemolymph, can be uptaked by the fat body cells of prepupa and then how the uptaked storage proteins can be accumulated in the fat body cells, if uptaken. The fat bodies which had been isolated from last instar larvae were cultured in 1 ml of Grace's insect medium containing $50{\mu}l$ of $^{3}H$-leucine (5.0 mCi/mol, Dupont) at $28{\pm}2^{\circ}C$ for 6 hrs. After the homogenates of the cultured fat bodies were centrifuged at 10,000 rpm for 10 minutes, the proteins included in the supernatant were separated by polyacrylamide gel electrophoreses (non-SDS, 6%). The next treatment of the electrophoresed gel was followed by rinsing. A storage protein band of several bands in the rinsed gel was sliced off. With elution of sliced storage protein bands in Tris-glycine buffer, the purification of radioactive storage proteins from fat bodies was finished. After the purified radioactive storage proteins were added in Grace's insect midis containing fat bodies of the prepupae, they were cultured for the randomly following minutes given as 3, 5, 7, 10, 15, 20 and 30 and for the randomly following hours given as 1, 2, 3 and 4 respectively. The double fixations of the cultured fat bodies in aldehyde and $OsO_4$, were followed by preparation of ultrathin sections from Epon-Araldite blocks through dehydration and embedding. The electron microscope autoradiographic treatment of all prepared sections were performed by the dipping method (Kim et al., 1987). The finally prepared specimens were examined with electron microscope. The fat body cells of the prepupa could be found to uptake the storage preteins of the last instar larvae, which were included in the culture medium, mostly by formation of coated vesicles. The in vitro uptake of the storage proteins actively occurred by 30 minutes after the addition of purified storage proteins in the culture medium. After culture for 7 minutes with the storage proteins, the uptaked radioactive storage proteins labelled a number of lysosomal granules. After culture for 20 minutes with the storage proteins, the radioactive storage proteins were finally incorporated and accumulated in lipid droplets and protein granules. The frequency in the fat body cell of radiolabelled lipid droplets occurs approximately 60%, while the frequency, in which the radiolabelled protein granules occurs in a fat body cell, is approximately 40%.
조정숙,김관선,김우갑,Cho, Jeong-Sook,Kim, Kwan-Seon,Kim, Woo-Kap 한국현미경학회 1991 Applied microscopy Vol.21 No.2
Ventral nephrocytes in the larva of the Lucilia illustris comprise ellipsoid cells situated onto the salivary glands. The cells are $60{\sim}100{\mu}m$ in diameter. Junctional complex beneath the basement membrane hold the plasma membrane in a even contour. Intracellular channels from the juntion complex are well developed at the cortex part of the cell. Coated vesicles pinched off from the channels seems to be connected with the ${\alpha}$-vacuoles via the tubular elements, which is regared as selective absorption system from the hemolymph. Two nuclei are sometimes observed in the medulla part of the cell. Ventral nephrocytes contain well-developed rough endoplasmic reticulum and Golgi complex, and numerous mitochondria. These cellular organelles synthesize lysosome. The lysosome not only digest some cell organells but also seems to be related with the ${\beta}$-vacuoles.
Melanophores of Pungitius sinensis and Pungitius kaibarae(Gasterosteidae, Plsces)
Youn, Chang-Ho,Lee, Sang-Dae,Kim, Kwan-Seon 한국어류학회 2002 韓國魚類學會誌 Vol.14 No.2
산란기와 비산란기로 구별하여 가시고기와 잔가시고기의 피부와 등지느러미 가시막에 있는 멜라닌색소포의 분포와 모양을 조사하였다. 비산란기의 가시고기와 잔가시고기는 큰 차이를 보이지 않았지만 가시고기의 등지느러미 가시막은 투명한 반면, 잔가시고기의 가시막은 흑색을 보였다. 산란기의 가시고기는 옅은 갈색으로의 체색 변화만 일으켰지만, 잔가시고기의 피부는 짙은 흑색으로 큰 변화를 보였다. 그러나 외형으로 나타나는 체색과는 달리 멜라닌 색소포의 수와 그 내부의 멜라노솜 수는 큰 차이를 보이지 않았다. 조면소포체와 골지체, 그리고 멜라닌세포의 분포도 두 종간에 차이가 나타나지 않았다. 특히 잔가시고기의 피부는 산란기와 비산란기에 큰 색깔 차이를 보임에도 불구하고 미세구조적인 소견에서는 큰 차이를 보이지 않았다. 옅은 색깔을 띠는 피부나 가시막의 멜라노솜은 구형으로 관찰되었으며, 짙은 흑색을 띠는 피부나 가시막의 멜라노솜은 불가사리 모양으로 세포질 돌기가 심하게 분지되어 있었다. 따라서 육안으로 나타나는 피부나 가시막의 색깔은 멜라노솜의 양보다는 그 모양이나 분포에 영향을 받는 것으로 나타났다. The distribution and structure of melanophores in the skin and membrane of the dorsal spines of the Pungitius sinensis and P. kaibarae were studied during breeding and non-breeding seasons. In the non-breeding season, the two fishes showed little difference except that the membrane of the dorsal spines of P. sinensis was transparent, while that of P. kaibarae was black. During the breeding season there were great differences between the two fishes in skin color: the skin of P. sinensis was light brown, while that of P. kaibarae was black. There was however, little differences between the two fishes in the number of melanophores and melanosomes of the skin. The distribution of the rough endoplasmic reticulum and Golgi complex, and the number of melanocytes also showed little difference in both species, especially both in breeding and non- breeding season in P. kaibarae. The melanophores in light-colored skin and in the membrane of the dorsal spines of P. sinensis were round, whereas the melanophores in the black-colored skin and in membrane of the dorsal spines of P. kaibarae had many projections of the cell processes, shaped like a starfish. Therefore, it is suggested that the color of the skin and the membrane of the dorsal spines depend not on the amount of melanosomes present but on the distribution of the melanosomes within the melanophores.