연두금파리 용화에 따른 타액선의 형태 변화

박의석,김관선문명진김우갑 한국통합생물학회 1992 동물학회지 Vol.35 No.3

화에 따른 연두금파리(Lucilia illustris Meigen) 타액선의 미세구조 변화와 타액선 내부에 함유된 분비물질의 형성과정 및 물질의 성분을 전자현미경 및 효소세포화학적 방법으로 관찰하였 다. 종령유충기의 타액선은 식도의 좌우에 위치한 한쌍의 반투명한 관으로 단층으로 배열된 편평형의 세포들로 이루어져 있었다. 세포질에는 조면소포체와 골지복합체가 풍부하였고, 세포내 물질의 생성이 매우 활발하였으며, 세포질내에 축적된 분비과립은 부분분비에 의해 배출되었다. 전용기의 타액선은 세포가 비후해짐에 따라 타액선의 내감이 줄어들며 타액선의 길이도 감소하였으며, 세포질내 조면소포체의 양적 감소와 대형분비과립의 형성이 현저하였다. 용기에는 대형의 자식성 액포들이 황범위하게 형성되어 있었으며 이는 유충시기의 타액선이 변태과정을 거치며 퇴화되는 것으로 사료되었다. 타액선 세포에 대한 acid phosphatase외 반응은 주로 내강면에 인접한 리소조옴에서 양성반응이 나타났으며, 종령유충 시기의 타액선 세포질내 지질성분은 주로 기저부에 존재하였으나, 용 형성 48시간 이후에는 관찰되지 않았다.

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%.

무당개구리(Bombina orientail Boulenger) 피부 과립선의 미세구조에 관한 연구

김기영,김관선,문명진,김우갑 한국통합생물학회 1989 동물학회지 Vol.32 No.2

무당개구리 피부과립선의 조직화학적 성질과 그 미세구조를 광학, 주사 및 투과형 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 무당개구리의 과립선은 진피에 위치한 단포상선으로서 선강재에 함유된 분비과립의 형태와 그 조직화학적 특성에 따라서 분비과립의 크기가 작은 I형과립선과, 분비과립이 큰 II형 과립선의 두 종류로 구분되었다. 이들은 공통적으로 점액선에 비해 선분비부가 잘 발달되어 있었으며, 내강이 없고 인접세포간의 세포경계가 관찰되지 않는 다핵체의 형태를 하고 있었다. 조직화학반응의 결과, 두 종류의 과립선은 Alcian blue에 전혀 반응하지 않았고, PAS반응에는 I형 과립선만이 약한 양성반응을 나타내었다. 또한 주사전자현미경으로 관찰한 과립선 개구부의 표면구조는 공통적으로 피부에 돌출된 기공과 흡사한 형태를 하고 있었는데, I형 과립선의 개구부는 미세한 망상구조를 가진 세포로 이루어져 있었고, II형 과립선은 비교적 큰 과립상의 표면구조를 가진 세포로 이루어져 있었다. 한편 과립선의 선분비부는 공통적으로 외부의 근상피세포층과 내부의 선상피세포층으로 되어 있었고, 선상피세포층내에 함유돈 각각의 분비과립들은 모두 조면소포체로부터 기원하였으며, 과립의 성숙정도에 따라서 다양한 형태적 변이가 관찰되었다. The histochemical characteristics of the cutaneous granular glands and its fine structure of the Korean fire bellied toad, Bombino orientalis Boulenger the only species inhabitated in Korea, were studied with light and electron microscopes. By the granular size and histochemical properties, the granular glands were subdivided into two types. The type I glands have small secretory granules (3.6 $\mu$ m), whereas the type II glands contain relatively larger granules (7.5 $\mu$ m). These two types of glands were simple alveolar gland, and they were commonly composed of inner glandular epithelial cells and outermost myoepithelial cells. By the histochemical reactions, these two secretory granules were not responsed to Alcian blue, but weakly positive reaction was observed in type I granular glands by the PAS reaction. And by the scanning electron microscopic observation, extemal morphology of the opening site of the gland duct was resembled to the stomata of the plant leaves. Especially the epithelial cells of the type I glands had fine reticular surface whereas, type II glands had large vesicular surface. These types of the glands conimonly had larger cavities than those of mucous glands, and forming the multinucleated unicellulr structure (syncytium) at the glandular epithelial layers. Secretory granules of these glands were originated from the rough endoplasmic reticulum of the glandular epithelial cells, and by the maturation level of the granule, morphological diversities appeared.

미국 흰불나방 유충과 용의 발생에 따른 전흉선(prothoracic gland) 의 호르몬 분필활성의 변화

이봉희,김지현,김관선,문명진,김창환,김우갑 한국곤충학회 1987 Korean journal of entomology Vol.17 No.2

An ultrastructural study on the prothoracic glands of fall-webworms (Hyphantria cunea Drury) was performed to reveal secretory activities of the ecdysone hormone in the five developmental stages of last-instar pupae, prepupae, pupae immediately after pupation, 3 -day-old pupae and diapausing pupae. The results obtained are as follows: The active prothoracic gland cells have well-developed smooth endoplasmic reticulum which produces the moderately electron-dense material which is presumed to be ecdysone hormone. The edysone, after being synthesized, is transported into vesicular structures formed at the marginal ends of the smooth tubular endoplasmic reticulum. Some vesicles produced by the separation of vesicular structures from the smooth tubular endoplasmic reticulum were incorporated into either other vesicles or smaller electron-lucent spheroids to form larger secretory electron-lucent spheroids. The spheroids containing the ecdysone are released into haemolymph. According to the ultrastructural characteristics associated with the secretory activities of the ecdysone, the prothoracic glands of the prepupae have high secretory activities, compared with the prothoracic glands of the last-instar larvae with the secretory activities which a few cells of their prothoracic glands have. But not only most of the prothoracic gland cells of the last-instar larve but also all the prothoracic gland cells of pupae just after pupation, 3-day-old pupae and diapausing pupae are inacitve in secretory activities. The inactive prothoracic gland cells have also more highly indented nuclei and more abundant glycogen granules to an extent of the stages of the pupae.