연령에 따른 흰쥐 망막의 기저복합체의 변화에 대한 미세구조적 연구

고정식 순천향대학교 1989 논문집 Vol.12 No.1

본 실험은 연령에 따른 흰쥐 망막 기저복합체의 미세구조적 변화를 관찰하기 위하여 시행하였다. 실험동물로는 생후 3개월, 12개월 및 30개월된 Fisher 344계 흰쥐를 각군에 4마리씩 사용하였다. 각 동물은 4% chloral hydrate 마취하에 안구를 적출하였다. 적출된 안구에서 초자체를 제거한 후, 망막후부중 일정부위의 망막을 절취하였다. 절취된 조직은 2.5% glutaraldehyde-1.5% paraformaldehyde고정액(0.1 M phosphate buffer, pH7.3) 에 1차 고정한 후, 1% osmium tetroxide 액(0.1 M phosphate buffer, pH 7.3) 에 2차 고정하였다. 고정된 조직은 탈수과정을 거쳐 araldite에 포매한 후 LKB-V ultratome으로 60-70nm 두께의 얇은 절편을 만들었다. 각 절편은 uranyl acetate와 lead citrate로 대조염색을 시행한 후 JEM 100CXⅡ 전자현미경으로 관찰하였다. 기저복합체를 구성하는 5 층의 두께 측정은 각군당 50 매씩의 사진에서 비스듬히 절단된 기저복합체로 인한 오차를 줄이기 위해, 각 사진에서 얇은 부분을 측정하였다. 결과는 다음과 같다. 1. 망막 기저복합체의 두께는 어린 흰쥐의 것은 474nm, 젊은 흰쥐의 것은 525nm 및 늙은 흰쥐의 것은 704nm로서 연령증가에 따라 현저히 두꺼워졌다. 2. 맥락막의 모세혈관 기저막의 두께는 어린 흰쥐의 것이 121nmduTsmsep 비하여 젊은 흰쥐와 늙은 흰쥐의 것은 각각 66nm와 54nm로서 연령이 증가함에 따라 얇아졌다. 3. 외측 교원섬유층의 두께는 어린 흰쥐의 것은 132nm, 젊은 흰쥐의 것은 139nm 및 늙은 흰쥐의 것은 232nm로서 연령이 증가함에 따라 두꺼워졌다. 4. 탄력섬유층의 두께는 어린 흰쥐의 것이 91nm였고, 젊은 흰쥐와 늙은 흰쥐의 것은 각각 147nm와 142nm로서 연령이 증가함에 따라 두꺼워졌다. 5. 내측 교원섬유층의 두께는 어린 흰쥐의 것이 87nm였고, 젊은 흰쥐와 늙은 흰쥐의 것은 각각 106nm 와 119nm로서 연령이 증가함에 따라 두꺼워졌다. 6. 색소상피 기저막의 두께는 어린 흰쥐의 것이 43nm였고, 젊은 흰쥐와 늙은 흰쥐의 것은 각각 67nm 와 157nm로서 현저히 두꺼워졌다. 7. 늙은 흰쥐의 경우, 색소상피 세포와 기저막 사이에 미세한 섬유상 또는 과립상물질<기저침전물>이 침전되어 있는 경우가 자주 관찰되었다. Morphological difference of the Bruch's membrane at different age groups have been studied in young (three months-old), adult(twelve months-old) and old(thirty months-old) Fisher strain 344 rats. Pieces of the tissues taken from the posterior regions of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH7.3) and 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH7.3), embedded in araldite mixture. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100 CX II electron microscope. The mean thickness of each of the five zones of the Bruch's membrane were determined by measuring the thinnest portion of Bruch's membrane, and by taking the average of 50 readings from electron micrographs at different ages. Observed results were as follows: 1. The thickness of the Bruch's membrane is increased during aging: 474 nm in young rats, 525 nm in the adult, and 704 nm in the old. 2. The thickness of the basal lamina of the choriocapillary endothelium is decreased during againg: 121 nm in young rats, 66 nm in the adult, and 54 nm in the old. 3. The thickness of the outer collagenous zone is increased during aging: 132 nm in young rats, 139 nm in the adult, and 232 nm in the old. 4. The thickness of the elastic zone is increased during aging: 91 nm in young rats, 147 nm in the adult, and 142 nm in the old. 5. The thickness of the inner collagenous zone is increased during aging: 87 nm in young rats, 106 nm in the adult, and 119 nm in the old. 6. The thickness of the basal lamina of the pigment epithelium is increased during aging: 43 nm in young rats, 67 nm in the adult, and 157 nm in the old. 7. Accumulaton of finely filamentous or granule-like materials(basal deposit) between basal foldings of pigment epithelium and above its thickened basal lamina is more frequently observed in old rats.

Mitomycin C가 생쥐 콩팥소체의 미세구조에 미치는 영향

고정식,안의태,박경호,박대균 순천향대학교 교수학습개발센터 2004 Journal of Soonchunhyang Medical Science Vol.10 No.3

The experiment was performed to study the morphological responses of the renal glomeruli of the mice after administration of mitomycin C. Healthy adult mice weighing about 25gm each were divided into normal and experimental groups. Mitomycin-C(400㎍/kg) was injected subcutaneously to the animals every other day, and animals were sacrificed at 4 days or 6 days following the first injections. Pieces of tissues were taken from renal corticies, prefixed with 2.5% glutaraldegyde- 1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.), and followed by post-fixation with 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH 7.3). Fixed blocks were dehydrated with alcohol and acetone, embedded in araldite mixture, cut with LKB V-ultrotome. The ultratihin sections stained with urany1 acetate and lead cirtrate were observed with a JEM 100CX-Ⅱ electron microscope. In the fourth and seventh day following the first injection of mitomycin C, components of the renal glomeruli of the mice are looked compact since they were filled with the widened the mesangium, and showed narrowing lumen of glomerular capillaries and of urinary spaces. The electron densities of the lamina rara interna and the lamina rara externa were similar to the density of the lamina densa and the expanded lamina rara interna were often seen. and the cisternae of the rough endoplasmic reticulum of the podocytes were slightly swollen, whereas flattened cisternae of the Golgi complexes were frequently observed. From the above results, it was suggested that although small amount of mitomycin C(400ug/Kg) may give damages on the fine structure of the renal glomeruli of the mice.

연령이 증가함에 따른 흰쥐 망막의 막대세포, 원뿔세포 및 바깥얼기층의 미세구조 변화

고정식,한종원,양남길,안의태 대한해부학회 1992 Anatomy & Cell Biology Vol.25 No.3

발꿈치힘줄이 절단된 흰쥐의 가자미근과 장단지근내의 모세혈관과 신경섬유의 형태학적 연구

고정식,최기수,양남길,안의태,박경호,김진국 대한해부학회 1995 Anatomy & Cell Biology Vol.28 No.1

연령에 따른 흰쥐 망막의 변화에 대한 형태학적 연구

고정식 순천향대학교 1989 논문집 Vol.12 No.1

본 실험은 연령에 따른 흰쥐망막의 형태학적 변화를 관찰하기 위하여 시행하였다. 실험동물로는 어린쥐(생후 3개월), 젊은쥐(생후 12개월) 및 늙은쥐(생후 30개월)로 구분하여 Fisher 344계 흰쥐를 사용하였으며 각 군에 4마리씩 사용하였다. 실험동물은 4% chloral hydrate로 마취한 상태에서 안구를 적출하였으며, 적출된 안구에서 초자체를 제거한 후 망막후부중 일정부위의 망막을 절취하였다. 절취된 조직은 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH7.3)액에 1차 고정한 후, 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH 7.3)액에 2차 고정하였다. 고정된 조직은 탈수과정을 거쳐 araldite mixture에 포매한 후 LKB-V ultratome으로 1㎛ 두께의 절편을 작성하여 toluidine blue 액으로 염색한 후 광학현미경으로 관찰하였다. 망막을 구성하는 각층의 두께는 미측계(micrometer)로 계측하였으며 외망상층과 간상체·추상체층에 출현하는 외핵층세포핵의 수는 망막길이 3.7mm에 출현하는 핵의 개수를 헤아렸다. 결과는 다음과 같다. 1. 어린흰쥐의 망막두께는 256㎛였으며 젊은흰쥐와 늙은흰쥐의 것은 각각 253㎛와 198㎛였다. 2. 망막을 구성하고 있는 8층 가운데서 색소상피층과 신경섬유층을 제외하고는 모두 연령이 증가함에 따라 그 두께가 얇아졌는데, 특히 외핵층의 두께가 가장 심하게 감소되었다. 3. 간상체·추상체층에 출현하는 외핵층의 세포핵은 어린 흰쥐의 경우 망막길이 3.7mm 당 1.1개였으며, 증가함에 따라 그 출현빈도가 뚜렷하게 증가되었다. 4. 외망상층에 출현하는 외핵층의 세포핵의 수는 어린흰쥐의 경우 망막길이 3.7mm 당 8.2개였으며, 젊은흰쥐는 27.8개, 늙은흰쥐는 65.7개로서 연령이 증가함에 따라 그 출현빈도가 뚜렷하게 증가되었다. To study the age-related morphological differences of retinae of rats, retinae of young(3 months-old), adult(12 months-old) and old(30 months-old) Fisher strain 344 rats were dissected out under anesthesia. Specific area from the posterior portion of retina of each rat was fixed in 2.5% glutaradehyde-1.5% paraformaldehyde, and postfixed in 1% osmium tetroxide. Fixed tissues were embedded in the araldite mixture, and 1 ㎛ thin sections with LKB-B ultratome. The sections were stained in toluidine blue solution. Under light microscope, thickness of retinal layers were compared with micrometer, while the ectopic nuclei from outer nuclear layer appearing in the rod and cone layer and in the outer plexiform layer were counted within the 3.7 mm-length of the retinae. The results were as follow: 1. Thickness of retinae of rats show differences according to their age. In the young rat, mean thickness of retina was 256㎛, while that of adult rat was 253㎛ and that of old rat was 198㎛, respectively. 2. Of the 8 retinal layers, thickness of each layer, except the pigment epithelium was thinned out. Thinning was most significant in the outer nuclear layer. 3. Ectopic nuclei scattered in the rod and cone layer were 1.1 in young rats, 3.5 in adult rats, 16.2 in old rats per each 3.7 mm-length of retinal sections, respectively. Data show the significant increase of ectopic nuclei in the rod and cone layer according to the aging of the rat. 4. Ectopic nuclei in the outer plexiform layer were 8.2 in young rats, 27.8 in adult rats, and 65.7 in old rats per each 3.7 mm-length of retinal samples, respectively. It shows also significant age-related increase of ectopic nuclei in the outer plexiform layer.

과량의 방사선에 머리부분이 노출된 흰쥐 망막의 속핵층과 신경절세포의 미세구조변화

고정식,최숙경,양남길,안의태,박경호 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.2

이 실험은 머리부분이 많은 양이 방사선에 노출되었을 때, 망막 속핵층과 신경세포의 미세구조적 반응을 관찰하기 위하여 시행하였다. 실험동물로는 체중200-250g의 흰쥐를 사용하였으며, 마취상태에서 머리부분에만 방사선을 조사였다. 방사선조사는 선형가속기(Mitsubishi Linear Accelerator, ML-4MV)를 이용하였으며 1분당 200rad씩 연속 조사하여 3,000rad 또는 6,000rad를 조사한 후 6시간, 2일, 6일에 희생시켰다. 각 동물은 1% glutaradehyde- 1% paraformaldehyde액으로 관류고정한 다음, 안구를 적출하여 톱니둘레를 따라 절단한 후 망막의 일정부위에서 조직을 떼어내었다. 떼어낸 조직은 2.5% glutaradehyde- 1.5% paraformaldehyde 액에 일차고정한 다음 1% osmium tetroxide액에 이차 고정하였다. 고정된 조직은 수세와 탈수과정을 거쳐 araldite혼합액에 포매한 다음 얇은 절편을 작성하여 uranyl acetate와 lead citrate로 염색한 후 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 무축삭신경원은 세포질의 전자밀도가 높은 어두운 세포(type 3)와 전자밀도가 낮은 밝은 세포로 구분되었으며, 밝은 세포는 핵의 윤곽이 매끄럽고 둥근 세포(type 1)와 핵막의 함입현상이 뚜렷한 세포(type 2)로 구분되었다. 2. 3,000rad의 X-선을 조사한 후 6시간군의 경우, 속핵층을 구성하는 세포들과 신경절세포에서는 속핵층을 구성하는 세포들과 신경절세포에서는 사립체와 세포막이 파괴되는 모습이 관찰되었는데 2일군의 경우 그 경향이 가장 심했다. 3. 3,000rad의 X-선을 조사한 후 6일군의 경우, 부챗살아교세포의 영역이 넓어져서 전체적인 모습이 2일군의 것에 비하여 많이 회복되어 보였다. 4. 6,000rad의 X-선을 조사한 후 6일군의 경우, 속핵층을 구성하는 세포들이 기저복합체 근처에서 관찰되었다. 5. 6,000rad의 X-선을 조사한 후 6시간부터, 부챗살아교세포는 미세사가 많고 전자밀도가 높은 변성된 세포들이 자주 관찰되었다. 이상의 결과를 고찰해 보면 머리부분이 3,000rad의 X-선에 노출되면 망막의 속핵층을 구성하는 세포들과 신경절세포는 처음에는 새포질소기관들과 세포막이 손상을 받으나, 6일 후에는 부챗살아교세포의 영역이 넓어져서 전체적인 구조가 많이 회복된다. 그러나 6,000rad정도의 방사선에 노출되면 부챗살아교세포도 변성되어 속핵층과 신경절세포층의 구조가 변성된다고 생각된다. This experiment was performed to study the morphological responses of the cell of the inner nuclear layer and the ganglion cell of the rat retina following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000rads or 6,000rads of radiation in a single dose, respectively. The source was a Mitsubishi linear accelerator ML-4MV. The target to skin distance was 80cm, and the dose rate was 200 rads/min. The experimental groups were sacrificed on 6hour, 2 and 6 days after X-ray irradiations. Under anesthesia, 1% glutaraldehyde-1% paraformaldehye solution was perfused through the heart. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, and in 1% osmium tetroxide, and embedded in araldite mixture. Ultrathin sections were stained with uranyl acetate and lead citrate solutions and were observed with JEM 100CX-II electron microscope. The results were as follow: 1. Three type of amacrine cells were found, according to their electron densities and nuclear shape, i,e,dark amacrine cell with invaginated nucleus(type 3), light amacrine cell with round nucleus(type 1) and light amacrine cell wit invaginated nucleus(type 2). 2. Six hours after exposure to 3,000 rads of X-ray, morphological changes were observed in cytoplasm of the cells of the inner nuclear layer and the ganglion cell: i.e., edematous changes, fragmentation of mitochondria, breakdown of the plasma membrane, etc. 3. Six days after exposure to 3,000rads of X-ray, many of the malformations were seen to be repaired, except the territory of activated Muller cell. 4. Six days after exposure to 6.000 rads of X-ray, cells of the inner nuclear layer were observed near the Bruch's membrane. 5. Six days after exposure to 6.000 rads of X-ray, darkly degenerated Muller cells were frequently observed. From the above results it wluded that 3,000rads of head-irradiation induces damages to the cell organelles in the cells of the inner nuclear layer and the ganglion cell during the first few days, but many of the malformations can be repaired within six days. And 6,000rads of head irradiation leads to renaturation of the inner nuclear layer and the ganglion cell layer of the retina, since the most of the Muller cells are degenerated within a few days following irradiation.

부신절제후의 위점막 점액세포의 미세구조

고정식,양남길,구광일,안의태,박경호 순천향대학교 1989 논문집 Vol.12 No.3

The experiment was conducted on the male Sprague-Dawley rats weighing about 250g each, to study the fine structural changes of the gastric mucus-secreting cells following bilateral adrenalectomy. Adrenalectomy was performed by dorsal approach method under ether anesthesia. Animals are fed ad libitum after the operation. The normal and the adrenalectomized rats were sacrificed on the 1st, 3rd, 5th and 7th day after the operation. For the electron microscopic observation, gastric mucosae were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde followed by post fixation in 1% osmium tetroxide. The ultrathin sections were contrasted with uranyl acetate and lead citrate, and examined with JEM 100CX Ⅱ electron microscope. Obtained results were follows: 1. The gastric surface mucous cells of adrenalectomized rat contained mucous granules of less electron density than those of the normal rat contains. And mucous granules of adrenalectomized rats are usually placed apart from the surface plasma membrane. 2. Goblet cells of adrenalectomized rat exhibited well formed microvilli as compared with those of normal rat. 3. Cell differentiation in the isthmic area was generally seem to be delayed following adrenalectomy. 4. Golgi complexes of goblet cells were influenced by adrenalectomy, showing flattening or disorganization of cisterns, non-typical vacuoles, scanty vesicles, etc. 5. Considering the above findings, adrenalectomy probably causes the dysfunction of the gastric mucus cell that results in delayed mucus formation and secretion.

Acriflavine-Guanosine 복합제 (AG60)의 조직내 분포에 대한 형광현미경적 연구

고정식,임수재,안의태,김진국,김상건,이경영,강종구,홍은경,정영신,유보림,한영복 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.1

X-선 조사를 받은 흰쥐망막 빛감각세포의 미세구조

고정식,박동분,양남길,안의태,박경호,김진국 대한해부학회 1993 Anatomy & Cell Biology Vol.26 No.3

집토끼의 갑상선내 Parafollicular Cells에 대한 전자현미경적 연구

고정식,양남길 순천향대학교 1981 논문집 Vol.4 No.2

In order to study the fine structure of the parafollicular cells in the rabbit thyroid gland, small b1ocks of the thyroid tissue were fixed in the mixture of 2% paraformaldehyde-2.5 % glutara1dehyde (phosphate buffer, pH 7.4) and post-fixed in 1 % osmium tetroxide(phosphate buffer, pH 7.4). The tissue were embedded in Epon 812, ultrathin-sectioned with the LKB-Ⅲ ultramicrotome. The sections contrasted with uranyl acetate and lead citrate were observed with a JEM 100B electron microscope. Observed results were as follow: 1. Of the rabbit parafollicular cells. the cells with electron-lucent cytoplasm (light cell) and with electron-dense cytoplasm (dark cell) can be distinguished. 2. The parafollicular cells of the rabbit were observed in the intra-. para-and inter-follicular positions. Inter-follicular cells occur as groups of a few cello. None of intra-follicular cells have contact with the follicular lumen. 3. The parafollicular cells have oval or round indented nuclei. They contain moderately developed cell organelles. such as Golgi complex, granular doplasmic reticulum, ribosomes, etc. 4. The membrane-bound granules are spherical with a diameter of 150∼380nm. The electron densities of the core texture are variable from the coarse granule with lower density to the homogeneous granule with higher dentsity.