• Applied Microscopy
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• 제22권2호
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• pp.30-45
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• 1992

An experimental study on the acute irradiation effects on the substantia nigra of head-irradiated rats were carried out. Rats anesthetized with sodium thiopental, were exposed only on their head areas with a single dose of 3,000 rads or 6,000 rads, respectively. Radiation was produced by Mitsubishi linear accelerator at the speed of 200 rads/min. Aminals were sacrificed on 6 hours, 2 days and 6 days following irradiations. By the perfusion fixation through the heart, rats were fixed with 1% glutaraldehyde-1% paraformaldehyde solution. Two hours later, brains were exposed and immersed in the same fixatives over night. Tissue blocks from subtantia nigra were punched out, and they were refixed in the 2% osmium tetroxide solution. Blocks were dehydrated through alcohol series, and embedded in the araldite mixture. Ultrathin sections were stained with uranyl acetate and lead citrate solutions, From the ultrastructural study, following results were made: 1. Six hours after irradiation, severe depletion of synaptic vesicles was occurred in the many axon terminals of the nigral neuropil. 2. Dramatical decrease of lysosomes and dense granules was observed. 3. Two days following irradiation, alterations of ribosomes, granular endoplasmic reticula, mitochondria, etc, were noticed. 4. Many of the malformations were seen to be repaired on the 6th day. 5. Above results were interpreted as follows. At the acute stage of heavy irradiation, neurotransmitters in the substantia nigra are released severely. But they are recovered within 6 days. It is concluded that acute head-irradiation may result severe disturbance of nigral motor control function during the first few days.

• 한국수정란이식학회지
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• 제25권4호
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• pp.237-245
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• 2010

Only an optimum number of viable spermatozoa in a frozen-thawed insemination dose can ensure conception at artificial insemination (AI). We report here the percentages of normal, abnormal and viable spermatozoa present in the frozen-thawed semen of 20 Black Bengal bucks used for commercial AI. Bucks in this experiment were of 19.3~46.1 months old and 25~42 kg body weight. Four semen straws (0.25 ml) from each buck were collected for evaluation of their kinetic parameters. Scrotal circumference was measured by using a scrotal tape, sperm motility was estimated on eye estimation and sperm concentration was determined by using a haemocytometer. Sperm morphology was studied in paraformaldehyde fixed spermatozoa under differential interference contrast (DIC) microscope. To determine the proportion of live (plasma membrane intact) spermatozoa, semen was stained with SYBR-14 and propidium iodide and examined under fluorescent microscope. Scrotal circumference, post-thaw sperm motility, sperm concentration per insemination dose and proportion of normal spermatozoa were $21.5{\pm}0.7\;cm$, $43.5 {\pm}5.4%$, $83.5{\pm}6.7$ million and $88.3{\pm}4.1%$, respectively. The percentages of spermatozoa with head shape and acrosome abnormalities were lower ($2.7{\pm}1.1$ and $1.4{\pm}1.3$, respectively), whereas higher percentages of abnormalities ($7.0{\pm}1.8$) were observed in mid piece and tail portion. The proportion of live spermatozoa was $28.5{\pm}5.4$. It is concluded that although a good number of morphologically normal spermatozoa are present in the insemination dose, the proportion of live spermatozoa is low, which warrants further improvements of buck semen freezing procedures to ensure good quality at AI.

• Applied Microscopy
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• 제26권4호
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• pp.431-446
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• 1996

This experiment was performed to study the ultrastructural changes of the juxtaglomerular cell of mice following subcutaneous injection of heavy metallic agents. Male mice were divided into normal and experimental groups. The mice were subcutaneouly injected with $HgCl_2$ (2mg, 5mg or 10 mg/Kg/BW) or with $K_{2}Cr_{2}O_7$(5 mg, 10 mg or 20 mg/Kg/BW). Mice were sacrificed on 6 hours, 3 days and 14 days after the injection. Kidneys were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture. The sections were cut on a LKB-V ultratome, and ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX II electron microscope. The results were as follow: 1. Juxtaglomerular cell of the experimental groups showed some alterations, especially in the structures of protein synthesis including dilations and degradations of granular endoplasmic reticula, atrophy of Golgi complex, and numerous free ribosomes in the cytoplasm. 2. Juxtaglomerular cells treated groups showed a number of vacuoles, protogranules and some myelin figures in the cytoplasm, especially in the earlier groups. 3. Juxtaglomerular cells of treated groups, contained a large number of secretory granules showing variable electron densities and pleomorphism in later groups (2 weeks). From the above results, it was concluded that, the mercuric chloride or potassium bichromate induces acute renin release from juxtaglomerular cells of the mice, but many juxtaglomerular cells may secrete prematured secretory granules, or the synthetic system of the cell can not perform normal function.

• 대한수의학회지
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• 제39권3호
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• pp.407-416
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• 1999

This study was carried out to investigate the NPY-immunohistochemical characteristics of the olfactory bulb in the striped field mouse(Apodemus agrarius). The animals were anesthesized with thiopental sodium and perfused with 4% paraformaldehyde through left ventricle and aorta. Brains were removed and tranfered 10%, 20% and 30% sucrose. Sections were then cut on a cryostat into $40{\mu}m$-thick. The tissue immunostained with avidin-biotinylated complex method. The main olfactory bulb consisted of seven circumferential laminae : an olfactory nerve fiber layer, a glomerular layer with glomeruli surrounding by periglomerular cells, an external plexiform layer having granule and tufted cells, a mitral cell layer, a narrow internal plexiform layer, a granule cell layer forming several cell rows and a layer of white matter. The accessory olfactory bulb had four layers : an olfactory or vomeronasal nerve fiber layer, a glomerular layer consisting of small glomeruli, a mixed layer not distinguishing the external plexiform/mitral cell/granule cell layers and a granule cell layer. Most of NPY-immunoreactive(NPY-IR) neurons in main olfactory bulb were localized in the deeper portion of granule cell layer, white matter and anterior olfactory nucleus. In addition, some NPY-IR neurons were identified in the external plexiform layer. The shape of NPY-IR neurons of all olfactory bulb were predominant round or oval, sometime multipolar in shape. And most NPY-IR processes were parallel to long axis of white matter. In accessory olfactory bulb, NPY-IR neurons were not found in all region.

• Journal of Acupuncture Research
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• 제22권4호
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• pp.55-63
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• 2005

목적 : 만성 염증성 질환에 대한 전침효과를 알아보기 위해 complete Freund's adjuvant (CA) 유발 관절염 모델에서 염증관련 단백질의 변화를 살펴보았다. 방법 : Sprague-Dawley계 흰쥐의 족부에 CFA를 주사한 다음 3일 간격으로 2 Hz, 15 Hz 및 120 Hz 전침 자극을 주며 부종 형성여부를 plethysmometer로 측정하여 판정하였으며 30일 째 거퇴관절을 취하여 4% paraformaldehyde에 고정하고 EDTA용액에서 탈회시켜 파라핀연속 절편을 얻어 $NF-{\kappa}B$를 비롯한 5종의 염증관련 단백질의 발현을 면역조직화학적으로 살펴보았다. 결과 : 관절연골내 면역반응 중 연골기질은 반응이 없거나 약하고 연골세포는 $NF-{\kappa}Bp65,\;I-{\kappa}B{\alpha},\;iNOS$반응이 강하며 특히 유리연골층에서 더 현저하였으나 염증 및 전침자극에 따른 변화는 없었다. 관절낭에서 면역반응을 살펴보면 염증유발시 활액세포의 면역반응세포는 $I-{\kappa}B{\alpha}$가 감소한 반면 iNOS, $IL-1{\beta}$는 증가하며 특히 iNOS 증가가 현저하였으며 전침자극에 의해 iNOS가 감소하였다. 활액막조직에서 모든 면역반응이 증가하며 특히 $NF-{\kappa}Bp65,\;I-{\kappa}B{\alpha},\;iNOS$ 반응이 현저한데 전침자극에 의해 $IL-1{\beta}$를 제외한 모든 반응이 감소하였다. 결론 : 만성 염증성 동물모델의 거퇴관절 내 염증관련 단백질은 관절연골보다 관절낭에서 큰 변화를 보이며 전침처치에 의해 이들 단백질 발현이 억제되는 것으로 보아 전침이 만성 염증성 질환에 효과적임을 알 수 있다.

• 대한치과의사협회지
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• 제10권5호
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• pp.307-316
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• 1972

신생 Ball/c Strain 종 웅성백서 28두를 사용하고, 28두중 14두는 실험군에, 나머지 14두는 대조군에 각각 배당하였다. 실험군과 대조군은 동물희생시간(30분, 1시간, 4시간, 8시간, 12시간, 24시간 48시간)에 따라서 7군으로 구분하고, 각군에 2두식 배정하였다. 실험군은 백서를 Bell Jar내에 넣고, 여기에 창소를 서서히 넣어 Westinghouse Oxygen Analyzer로 30내지 40ppm이 되도록 하고 15분 동안 방치한 후, 이를 밖으로 제거하여 신선한 공기를 5분동안 마시게 하였다. 이런 과정을 반복하여 결국 55분 동안에 45분간 무 산소 상태에 있게 하였다. 악하선의 소편을 2% Paraformaldehyde(0.1M Cacodylate fuffer) 용액에 4시간동안 고정하였고, 탈수후 전현 표본포리법에 따라 Epoxy유지를 혼합하여 이몰하였다. LKB Ultratome으로 1μ의 연속절편을 만들었고 야기시킨후 신생백서의 악하선세포에 세포학적변화를 보고저하였고, 기결과를 요약하면 다음과 같다. 1. 신생 백서에 산소결핍을 야기시켰을때에 나타나는 초기의 세포학적변화는 이를 이르킨 후 1시간과 4시간사이가 절정에 달하였다. 이실험에서 유해적 영향은 공포화하고 점액성 및 장액성세포의 핵이 농축변성하며 장액성분필과기의 방출이 지연되어 세포질첨단에 축적되었다. 이들의 회복은 적어도 4시후에 시작되과, 12시간에는 세포질의 대부분, mrgl 핵의 용태가 정상으로 되돌아간다. 24시간에는 실험군과 대조군간에 있어서 야간의 차이는 인정되나 선소엽은 아직 초기에 볼수있었든 퇴행성변화의 잔여물을 함유하고 있었다. 2. 위에 요약한 본연구의 결과는 노인들이 보고한 산소결핍증이 악하선에 있어서도 단백질합성의 억제현상을 나타냈음을 세포학적으로 확증한 것이라하겠다.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.51-58
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• 1998

본 연구는 체외수정 프로그램에 참여하는 환자에 있어서 난자회수 이틀째의 자궁내막의 발달상태를 알아보기 위하여 pinopode의 발달상태, 에스트로젠 및 프로제스테론 수용체의 발현을 관찰하였다. 생검한 자궁내 막 조직 을 양분하여, 절반은 전사전자 현미경 (scanning electron microscope)으로 pinopode를 관찰하기 위하여 2.5% glutaraldehyde와 2% paraformaldehyde로 고정하였고, 나머지 절반은 dating 및 스테로이드 수용체의 면역조직화학적 측정 (immunocytochemistry)을 위하여 10% formalin으로 고정하였다. 모두 12명의 환자중 8명에서 pinopode가 관찰되었으며, pinopode 발달이 관찰되지 않은 환자들은 hCG 주사를 맞는 날의 estradiol (E2)의 혈중농도가 600 pg/mL이하로 낮았다. 본 연구의 결과로부터 자궁내막의 발달상태를 알아보기 위해서는 지금까지 일반적으로 사용되어 오던 dating이나 스테로이드 수용체의 면역조직화학적 측정법 이외에도 pinopode를 관찰함으로써 조금 더 정확한 진단을 할 수 있으리라고 사료되며, pinopode의 발달은 E2의 혈중농도와 관계가 있을 것으로 추정된다.

• Applied Microscopy
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• 제15권2호
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• pp.98-110
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• 1985

This study was performed to clarify the morphological structures of the epithelia of the renal papilla, renal pelvis and ureter of the sheep (Ovis aries L.) through the light and scanning electron microscopes, Tissue specimens were taken from the renal papilla (common renal papilla and peripelvic column) and the renal pelvis (pelvis proper and pelvic pouch) of the kidney and the ureter. For the light microscopy, tissue blocks were fixed in 10 % neutral buffered formalin and embedded in paraffin wax, serially sectioned at a thickness of $6{\mu}m$. These sections were stained with hematoxylin-eosin and periodic acid-Schiff reaction. For the scanning electron microscopy, tissue blocks were prefixed in 1% glutaral-dehyde-1.5% paraformaldehyde solution and postfixed in 1% osmium tetroxide solution, dehydrated in graded alcohol, transferred to isoamyl acetate, and then dried by the critical point dryer (Polaron E 3000). These dried tissues were coated with gold and observed with a scanning electron microscope (JSM-35C), The results were as follows: The apex of the common renal papilla was lined with simple columnar epithelium having many microvilli on its luminal surface. Lateral portion of the papilla was lined with stratified epithelium $2{\sim}3$ layers thick, and its superficial cells were microvillar cells having many microvilli. The epithelium lining the peripelvic column was $1{\sim}2$ layers thick. The superficial layer was made of the microvillar cells, but a few microplica cells were appeared in the region near the pelvic pouch. The epithelium of the pelvic pouch was $1{\sim}2$ layered transitional type, and its superficial cells were microplica cells. The epithelia of the pelvis proper and ureter were $4{\sim}6$ layered transitional type, and their superficial cells were typical facet cells existing many round depressions and ridges of cell membranes of the luminal side.

• 대한수의학회지
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• 제42권1호
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• pp.7-20
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• 2002

This study deals with the distribution and morphological study of the neurons with the neuropeptide Y (NPY) in developing chick brain. The developing brains of Korean native chicks at embryonic days 8 (E8), E10, E12, E14, E16, E18, and E20 were used. The chicks were perfused with 4% paraformaldehyde through the left ventricle and aorta. The brains were removed and transferred into 30% sucrose, and then cut in a cryostat into $60{\mu}m$ in thickness. The sections were immunostained with free-floating and avidin-biotin complex (ABC) methods. The numerous neurons with NPY were first observed in nucleus rotundus of diencephalon at E8. In particular, neurons in nucleus rotundus had the well-developed processes. At E12, the neurons with NPY were distributed widely in diencephalon; nucleus septalis lateralis, medialis, nucleus magnocelluaris preopticus dorsalis, medialis, ventralis, nucleus preopticus medialis, nucleus dorsolateralis anterior thalami pars magnocellularis, and nucleus paraventricularis magnocellularis (paraventricular nucleus) except nucleus rotundus. From E12 to E20, positive neurons were oval-shaped, changed gradually into the spherical- and multipolar-shaped. The shapes of processes were also omnidirectional and the number of those were less than in telencephalon. As the chicks developed, the morphology of neurons with NPY showed the tendency to increase in their sizes and numbers.

• 농업과학연구
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• 제48권3호
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• pp.607-615
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• 2021

Cytokines are protein mediators that possess the ability to assist cell-to-cell communication in immune system-related activities. In general, pathogen endotoxins activate the release of inflammatory mediators, and with time, there is an increase in the cytokine levels in the body. Interleukin (IL)-6 mediates the acute-phase inflammatory response, and elevated IL-6 levels have been reported in peritoneal fluids of women with pelvic inflammation and endometriosis, thereby associating it with oocyte quality and infertility. To overcome subfertility or infertility in humans and animals, the present study was done to examine the effect of recombinant IL-6 on porcine oocytes matured in vitro and subsequently to determine the fertilization rate and embryo development. Porcine oocytes were incubated with varying concentrations of IL-6 (0 - 2 ㎍·mL^-1) for 44 h followed by in vitro fertilization and culturing of the oocytes. The oocytes or embryos were fixed with 3.7% paraformaldehyde (PFA) and stained with fluorescence dyes, and the meiotic spindle, chromosome organization, fertilization status and embryo development were subsequently assessed under a fluorescence microscope. We observed induction of an abnormal meiotic spindle alignment in the oocytes incubated with IL-6 compared to the control oocytes incubated without IL-6. Moreover, significantly decreased fertilization rates and embryo development were observed for oocytes incubated with IL-6 (p < 0.05). Thus, an increased IL-6 level during oocyte maturation could be associated with fertilization failure due to an aberrant chromosomal alignment and a disruption of the cortical granules. Taken together, our results indicate that successful assisted reproduction can be achieved by controlling the levels of inflammatory cytokines.