• 한국임상수의학회지
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• 제30권6호
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• pp.442-448
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• 2013

개 정액 동결을 위한 glucose가 첨가된 glycerol-free TRIS 희석액을 개발하기 위해 glycerol-free TRIS내 알맞은 glucose의 양과 0.3 M glucose가 첨가된 glycerol-free TRIS내 정자의 적정 노출시간을 조사하였다. 여섯 마리의 수캐의 사출액을 0.04 M glucose가 첨가된 glycerol-free TRIS내에서 $4^{\circ}C$까지 100분 동안 냉각한 후 서로 다른 glucose농도 (0 M, 0,04 M, 0.1 M, 0.2 M, 0.3 M)의 glycerol-free TRIS에서 30분 동안 냉각하여 동결하였다. $37^{\circ}C$에서 25 초 동안 융해한 후 정자의 막 고유성과 첨단체 고유성을 검사하였다. 부가적으로 0.3 M glucose가 첨가된 glycerol-free TRIS내 정자의 적정 노출시간에 따른 정자의 동결 후 운동성, 생존성, DNA 고유성을 확인하였다. 막 고유성과 첨단체 고유성은 각각 6-carboxyfluoresceindiacetate(6-CFDA)/propidium iodide(PI) fluorescent staining와 Pisum sativum agglutinin conjugated-fluorescein isothiocyanate 방법에 의해 검사하였다. DNA 고유성은 terminal deoxynucleotidyl transferase dUTP nick end labeling로 염색하여 flow cytometry로 검사하였다. 0.2 M 또는 0.3 M glucose가 첨가된 glycerol-free TRIS에서 동결된 정자가 낮은 농도의 glucose가 첨가된 희석액에서 동결된 정자보다 막 고유성이 높게 나타났으며(p<0.05), 첨단체 고유성은 0.3 M 군에서 높게 나타났다(p<0.05). 운동성은 50 분 군에서 높게 나타났으나(p<0.05), DNA fragmentation index는 노출시간에 따라 차이가 없었다. 본 연구 결과 개정자가 0.3 M glucose가 첨가된 glycerol-free TRIS에서 $4^{\circ}C$, 50 분간 냉각 후 동결과 융해 후 더 높은 생존성을 나타냈다.

• Applied Microscopy
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• 제7권1호
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.407-417
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• 1999

Objective: Mammalian follicle cells are the most important somatic cells which help oocytes grow, mature and ovulate and thus are believed to provide oocytes with various functional and structural components. In the present study we have examined whether cumulus or granulosa cells might playa role in establishing the plasma membrane structure of mouse oocytes during meiotic maturation. Design: In particular the differential resistances of mouse oocytes against chymotrypsin treatment were examined following culture with or without cumulus or granulosa cells, or in these cell-conditioned media. Results: When mouse denuded oocytes, freed from their surrounding cumulus cells, were cultured in vitro for $17{\sim}18hr$ and then treated with 1% chymotrypsin, half of the oocytes underwent degeneration within 37.5 min ($t_{50}=37.5{\pm}7.5min$) after the treatment. In contrast cumulus-enclosed oocytes showed $t_{50}=207.0$. Similarly, when oocytes were co-cultured with cumulus cells which were not associated with the oocytes but present in the same medium, the $t_{50}$ of co-cultured oocytes was $177.5{\pm}13.1min$. Furthermore, when oocytes were cultured in the cumulus cell-conditioned medium, $t_{50}$ of these oocytes was $190.0{\pm}10.8min$ whereas $t_{50}$ of the oocytes cultured in M16 alone was $25.5{\pm}2.9min$. Granulosa cell-conditioned medium also increased the resistance of oocytes against chymotrypsin treatment such that $t_{50}$ of oocytes cultured in granulosa cell-conditioned medium was $152.5{\pm}19.0min$ while that of oocytes cultured in M16 alone was $70.0{\pm}8.2min$. To see what molecular components of follicle cell-conditioned medium are involved in the above effects, the granulosa cell-conditioned medium was separated into two fractions by using Microcon-10 membrane filter having a 10 kDa cut-off range. When denuded oocytes were cultured in medium containing the retentate, $t_{50}$ of the oocytes was $70.0{\pm}10.5min$. In contrast, $t_{50}$ of the denuded oocytes cultured in medium containing the filtrate was $142.0{\pm}26.5min$. $T_{50}$ of denuded oocytes cultured in medium containing both retentate and filtrate was $188.0{\pm}13.6min$. However, $t_{50}$ of denuded oocytes cultured in M16 alone was $70.0{\pm}11.0min$ and that of oocytes cultured in whole granulosa cell-conditioned medium was $156.0{\pm}27.9min$. When surface membrane proteins of oocytes were electrophoretically analyzed, no difference was found between the protein profiles of oocytes cultured in M16 alone and of those cultured in the filtrate. Conclusions: Based upon these results, it is concluded that mouse follicle cells secrete a factor(s) which enhance the resistance of mouse oocytes against a proteolytic enzyme treatment. The factor appears to be a small molecules having a molecular weight less than 10 kDa.

• The Korean Journal of Physiology
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• 제7권1호
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• pp.23-31
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• 1973

Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

• 농업과학연구
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• 제9권1호
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• pp.284-302
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• 1982

식량(食糧)으로서 단백질자원(蛋白質資源)의 개발문제(開發問題)는 동남(東南)아시아에 있어서 특(特)히 중요(重要)하며 한국(韓國)에 있어서도 섭취량(攝取量)이 부족(不足)한 현상(現狀)이다. 이 문제(問題)를 개선(改善)하는 방법으로 특(特)히 영양가(營養價)가 높은 동물성(動物性) 단백질(蛋白質)의 증산(增産)을 목적(目的)으로 한국재래산양(韓國在來山羊)의 사육(飼育)에 관(關)해서 연구(硏究)를 행하였다. 이 실험(實驗)은 우선 한국고유(韓國固有)의 재래산양(在來山羊)을 육자원(肉資源)으로 개발(開發)할 가치(價値)가 있는가를 확인(確認)하기 위해서 소형(小型) 제(第)1위법(胃法)으로 탄수화물(炭水化物)과 휘발성지방산(揮發性脂肪酸)의 흡수(吸收)를 실험(實驗)하였고 한편 in vivo법(法)에 의해서 제(第)1위(胃)의 아미노산 흡수(吸收)에 관해서 실험(實驗)을 했다. 이와 같은 실험(實驗) 결과(結果)에서 한국재래산양(韓國在來山羊)의 사육(飼育) 개선(改善)에 관해서 요약(要約)하면 다음과 같다. 1. 소형(小型) 제(第)1위법(胃法)에 의해서 재래산양(在來山羊)의 영양흡수(營養吸收)에 관하여 실험(實驗)한 결과(結果)는 영양소(營養素)를 주입(注入) 한 후 0.5, 1, 2시간(時間)의 간격(間隔)으로 측정(測定)한 V F A의 흡수율(吸收率)이 푸로피온산 70~86%, 초산 74~87%, 낙산 76~89%였다. 유기물(有機物)의 흡수율(吸收率)은 0.5%의 에칠 알콜이 29~89%였으며, 0.1M의 젖산이 12~27% 흡수(吸收)되었다. 2. 제(第)1위내(胃內)에 유리(遊離) L형(型) 아미노산을 투여(投與)하고 그 흡수(吸收)를 측정(測定)한 결과(結果)는 제(第)1위(胃) 정맥(靜脈)의 아미노산 함량(含量)에 있어서 전요(全要)아미노산구(區)에 비하여 메치오닌 결핍구(缺乏區)의 흡수율(吸收率)이 적었고 메치오닌 3배구(倍區)와 요소첨가구(尿素添加區)의 아미노산 흡수율(吸收率)이 높고 특(特)히 메치오닌을 결핍(缺乏)시켜도 점막층(粘膜層)의 함량(含量)에는 변화(變化)가 없었다. 3. 제(第)1위내(胃內)에 근육층(筋肉層)의 아미노산 흡수(吸收)도 점막(粘膜)과 같은 경향(傾向)을 보였으며 메치오닌 결핍(缺乏)에 의한 영향은 없고, 메치오닌을 3배(倍)로 첨가(添加)하면 전요(全要)아미노산구(區)에 비(比)하여 메치오닌과 글루타민의 함량이 147.44배(倍) 이상으로 증가(增加)되며 또 요소첨가구(尿素添加區)는 글루타민, 프롤린, 발린의 흡수율(吸收率)이 증가(增加)되었다. 4. 제(第)1위(胃) 정맥혈장(靜脈血漿) 중의 아미노산 농도(濃度)는 제(第)1위(胃) 점막(粘膜) 중이나 근육(筋肉) 중에서 보다 일반적(一般的)으로 낮으며 메치오닌의 결핍(缺乏)에 의해서 아미노산의 흡수율(吸收率)이 저하(低下)되고 메치오닌 3배구(倍區)나 요소첨가구(尿素添加區)는 아미노산 흡수(吸收)를 증가(增加)시키는 등 아미노산 조성분(組成分)에 따라서 그 흡수상태(吸收狀態)에 변화가 생긴다. 5. 요소첨가구(尿素添加區)는 제(第)1위(胃) 근육층(筋肉層)의 암모니아태질소(態窒素)와 아미노태질소(態窒素) 및 요소(尿素)의 함량(含量)을 증가(增加)시켰다. 재래산양(在來山羊) 제(第)1위(胃)안을 완전(完全)하게 깨끗이 할 수 없으므로 남아있는 세균(細菌)을 조사(調査)하였으며 암모니아태질소(態窒素)의 변화(變化)는 이러한 조건(條件)의 영향을 받았다. 6. 아미노산의 흡수차(吸收差)에 의한 제(第)1위(胃) 조직(組織)의 변화(變化)를 일반(一般) 현미경(顯微鏡)과 형광(螢光) 현미경(顯微鏡)으로 관찰(觀察)한 결과(結果)는 메치오닌 3배구(倍區)에 있어서 점막(粘膜)의 단층엔주상피(單層円柱上皮)와 고유층(固有層)이 약간 엷어졌으며 요소첨가구(尿素添加區)는 두껍게 변화(變化)하였는데 점막하조직(粘膜下組織)이나 근층(筋層)에 있어서는 각(各) 구간(區間)에 큰 차이(差異)가 인정(認定)되지 않았다.

• 한국임상수의학회지
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• 제24권4호
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• pp.577-583
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• 2007

It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

• International Journal of Oral Biology
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• 제33권1호
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• pp.33-43
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• 2008

In the process of bone remodeling, mineral phase of bone is dissolved by osteoclasts, resulting in elevation of calcium concentration in micro-environment. This study was performed to explore the effect of high extracellular calcium ($Ca{^{2+}}_e$) on mineralized nodule formation and on the expression of progressive ankylosis (Ank), plasma cell membrane glycoprotein-1 (PC-1) and osteopontin by primary cultured mouse calvarial cells. Osteoblastic differentiation and mineralized nodule formation was induced by culture of mouse calvarial cells in osteoblast differentiation medium containing ascorbic acid and ${\beta}$-glycerophosphate. Although Ank, PC-1 and osteopontin are well known inhibitors of mineralization, expression of these genes were induced at the later stage of osteoblast differentiation during when expression of osteocalcin, a late marker gene of osteoblast differentiation, was induced and mineralization was actively progressing. High $Ca{^{2+}}_e$(10 mM) treatment highly enhanced mRNA expression of Ank, PC-1 and osteopontin in the late stage of osteoblast differentiation but not in the early stage. Inhibition of p44/42 MAPK activation but not that of protein kinase C suppressed high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin. When high $Ca{^{2+}}_e$(5 mM or 10 mM) was present in culture medium during when mineral deposition was actively progressing, matrix calcifiation was significantly increased by high $Ca{^{2+}}_e$. This stimulatory effect was abolished by pyrophosphate (5 mM) or levamisole (0.1-0.5 mM), an alkaline phosphatase inhibitor. In addition, probenecid (2mM), an inhibitor of Ank, suppressed matrix calcification in both control and high $Ca{^{2+}}_{e^-}$treated group, suggesting the possible role of Ank in matrix calcification by osteoblasts. Taken together, these results showed that high $Ca{^{2+}}_e$ stimulates expression of Ank, PC-1 and osteopontin as well as matrix calcification in late differentiation stage of osteoblasts and that p44/42 MAPK activation is involved in high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin.

• 생명과학회지
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• 제20권4호
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• pp.467-473
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• 2010

갱글리오시드는 포유동물 세포막의 중요한 구성요소로서 세포와 세포 혹은 세포와 단백질간의 상호작용을 포함한 다양한 면역학적 역할을 수행하고 있다. 이 연구는 NIH-미니돼지의 간과 심장을 인간에게 이식할려고 할 때 예측되어지는 거부 반응과 관련된 구성성분들 중 시알산을 함유하고 있는 스핑고당지질인 갱글리오시드에 대해 조사하였다. 얇은막크로마토그래피와 면역조직화학적분석을 실시한 결과 NIH-미니돼지의 간은 갱글리오시드의 발현이 심장보다 높게 나타났다. 갱글리오시드 GD3, GD1a, GD1b, GT1b는 간과 심장의 두 기관에서 발견되었다. 그러나 GQ1b는 간에서만 발견되었고 심장에서는 검출되지 않았다. 이러한 결과는 갱글리오시드의 발현양상은 간과 심장에서 조직특이적이라는 것을 의미한다. 한편, GM3를 포함한 다른 갱글리오 시리즈인 갱글리오시드들은 NIH-미니돼지의 간과 심장에서 검출되어지지 않았다. 이와 같은 연구결과로부터 갱글리오시드는 미니돼지의 장기중 특히, 간과 심장의 이종장기이식과 관련된 면역거부반응에서 어떤 역할을 수행하고 있다고 여겨진다.

• The Korean Journal of Physiology
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• 제23권2호
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• pp.329-337
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• 1989

Cardiac muscles show stimulation frequency-dependent tension changes i.e. Bowditch phenomenon and Woodworth phenomenon, the former is an increase of tension with the increase of stimulation frequency, whereas the latter is an increase of tension with a decrease of stimulation frequency. Bowditch phenomenon is seen in the range of frequency 1.0 cps and above, and Woodworth phenomenon below the frequency 1.0 cps in the most of mammalian cardiac atrium. To throw some light on the possible mechanism of both phenomena in rat atrium, influences of drugs affecting $Ca^{2+}$ influx through the plasma membrane $(verapamil,\;La^{3+},\;norepinephrine)$ and $Ca^{2+}$ release from sarcoplasmic reticulum (SR) on frequency-tension curve were studied. The results obtained are summarized as follows: 1) At low temperature $(27.5^{\circ}C)$, both Bowditch and Woodworth phenomenon were demonstrated. But Bowditch phenomenon disappeared at the temperature above $(32.5^{\circ}C)$. 2) At $(27.5^{\circ}C)$, in the presence of verapamil, a $Ca^{2+}$ channel blocker, a time course of change in the frequency-tension was studied. It was found that Bowditch phenomenon was affected before the Woodworth phenomenon, then the former was completely disappeared. At $(32.5^{\circ}C)$, where no Bow-ditch is seen in normal atrial muscle, Bowditch phenomenon was reappeared by an administration of norepinephrine suggesting again that slow inward current of such as $Ca^{2+}$ channel is closely related to Bowditch phenomenon. 3) At $27.5^{\circ}C$, in the presence of $La^{3+}$, although tensions were decreased at all stimulation frequencies, Bowditch and Woodworth phenomenon were still demonstrated. However in the presence of both $La^{3+}$ and verapamil, Bowditch phenomena was disappeared suggesting that $La^{3+}$ is less effective in blocking $Ca^{2+}$ channel than verapamil. 4) At $27.5^{\circ}C$, in the presence of ryanodine, an inhibitor of calcium release from SR, Woodworth phenomenon was disappeared, which was consistent with previous reports of others, suggesting that $Ca^{2+}$ release from SR is closely related to Woodworth phenomenon. From the above findings, it may be concluded that Bowditch phenomenon is dependent on the magnitude of $Ca^{2+}$ influx through slow channel and Woodworth phenomenon is dependent on the amount of $Ca^{2+}$ stored in SR.

• The Korean Journal of Physiology
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• 제1권1호
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• pp.57-66
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• 1967

Cold $(0^{\circ}C)$ or warm $(25^{\circ}C)$ fresh and sea water were flooded into the lungs of rabbits through tracheal canule. Respiratory arrest ensued in 19.5 minutes in the warm fresh water flooded rabbits and was the longest survival time among the experimental groups. The survival times in the other groups were: 2.32 minutes in cold fresh water group, 2.75 minutes in .warm sea water group, and 4.57 minutes in cold sea water group. Cardiac output was measured by means of T-1824 dilution technique after 2 or 3 minutes of flooding in 27 rabbits. Blood pressure was observed by mercury manometer throughout the survival time in 40 rabbits. The following results were obtained. 1. Cardiac output in the warm fresh water flooded and sea water flooded animal was smaller than that of control rabbits. In the cold fresh water flooded animal cardiac output was greater than that of the control animal. 2. Time constants of T-1824 dilution curve of experimental group were elongated than the normal curve. 3. Central blood volume showed an increase in the fresh water group, a decrease in cold sea water group and no change in warm sea water group. 4. In all of the experimental groups arterial blood Pressure showed an abrupt and great variations after flooding of lungs and lasted about 30 seconds. Thereafter, arterial pressure remained at a plateau level until the sudden fall to zero and this was almost coincided with the time of respiratory arrest. The Plateau level of arterial Pressure in fresh water group was about 10 mmHg higher than the control value, and it was lower than the control value in warm sea water group. In cold sea water group the plateau was made up by fluctuations around the control value. 5. Osmosis of water through the lung alveolar membrane occured in all animals. Fresh water caused hemodilution and sea water caused hemoconcentration. 6. In sea water flooded animal more volume of water was recovered through the tracheal canule than the volume injected into trachea. This was interpreted as the consequence of the shift of water from plasma to alveolar sac. 7. Relative freight of lung was greater in fresh water group than sea water group. In all animal lung edema ensued. 8. The mechanisms of cardiac output variations were discussed.