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The Effect of Oocyte Activation on Development of Porcine Cloned Embryos

  • Kim, Y.S.;Lee, S. L.;Park, G. J.;S. Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.124-124
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    • 2003
  • The successful development of embryos cloned by nuclear transfer (NT)have been dependent on a wide range of known factors including cell cycle of donor and recipient ooplast, oocyte quality, NT procedure and oocyte activation. The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from 26 mm follicles of slaughterhouse ovaries and cultured for 22 h in NCSU #23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 g/mL LH, 0.5 g/mL FSH and 10 ng/mL EGF. The COCs were further cultured for an additional 22 h in the same medium at $39{\cird}C$ in an atmosphere of 5% $CO_2$ in air, without hormonal supplements. Primary cultures of fibroblasts isolated from a female fetus on day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM +0.5% FCS for 5 days in order to arrest the cells in G0/Gl. After enucleation, oocytes were reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 KV/cm, 30 sec) in 0.28 M mannitol containing 0.01 mM $CaCl_2$ and 0.01 mM $MgCl_2$. Eggs were then divided into three treatment groups, control (without further treatment, Group 1), eggs cultured in 10 g/ml cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 I drops of NCSU#23 supplemented with 4mg/ml BSA (essentially fatty acid free) until day 7 at $39{\circ}C$ in a humidified atmosphere of 5% $CO_2$. On day 4 the culture were fed by adding 20 I NCSU #23 supplemented with 10% FBS. Development rates into blastocysts were significantly higher (P<0.05) in Group 3 embryos compared to Group 1 controls ($27.6 \mu 2.7% vs. 20.1 \mu 4.1%$, respectively), but rates did not differ in Group 2 compared to control ($23.8 \mu 5.7%$). Total cell number in Group 3 blastocysts was however significantly higher (P<0.05) than in Groups 1 and 2 ($44.6 \mu 2.4 vs. 19.9 \mu 1.9 and 21.9 \mu 2.1$, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos.

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A Study on the Vegetables Mentioned in the Bible (성서에 언급된 채소류에 관한 연구)

  • Woo, Ja-Won
    • Journal of the Korean Society of Food Culture
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    • v.22 no.1
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    • pp.1-9
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    • 2007
  • This paper is intended to study what kinds of the vegetables are mentioned in the Bible and how they were used in those days. While one hundred and twenty-eight different plants are mentioned in the Bible, there are today 2,384 plant species in modem Israel, most of which have been introduced in recent centuries. These plants obviously did not exist there in biblical times and were only recently introduced from Australia and South America, respectively. This article will study only the vegetables mentioned in the Bible and known to have existed in the old and new testament times. Since the first book devoted exclusively to biblical botany was that of Levinus Lemmens in 1566, the modem systematic study of biblical plants, began with F. Hasselquist, a student of Linnaeus, the founder of modem botany. In 1928, Immanuel Loew approached the subject differently, reviewing all known data pertaining to biblical plants. His work not only discussed biblical plants, but also plants in later Jewish literature, particularly the Talmud. The British scholar G. E. Post provided a broad field study of modem plants in Palestine, Syria, Lebanon, and Jordan. More recent major treatments of the subject include those of A. and H. Moldenke (1952), M. Zohary (1982), N. Hareuveni (1984), and Y. J. Choi(1996). Today, articles on specific biblical plants listed in the Bible can be found in any number of encyclopedias. This study attempts to provide a synthesis of the work of a number of scholars who studied the vegetables and plants mentioned in the Bible. As a preliminary study on the culture of food in the biblical period, this study has focused on the identity and features of the vegetables of the Bible. In only a limited number of instances, because of the paucity of the informations and the broad and generic descriptions of the plants, we can't be certain about the identification of the vegetables named in the Bible. In many instances the traditions established by the Greek, Aramaic, and English translations are helpful, although sometimes they are misleading. This paper subdivides the vegetables into broad areas, the general vegetables and the flavoring herbs. Vegetables formed very important part of the diet in the biblical times. Two main types were used: those whose nutritious seeds could be easily stored and those which were eaten freshly gathered from gardens. Pulse seeds provided a useful source of vegetable protein, while fresh green vegetables were vitamin rich. Pulses could be eaten boiled, or their dried seeds could be ground up into flour and then made into nutritious soups. Fresh vegetables were eaten either raw or lightly cooked, usually by boiling in water. The general vegetables in the Bible are herbs(garden rocket), cucumber(snake cucumber), watermelon, leeks, chicory, and onions. Also the flavoring herbs in the Bible are rue, dill, cummin, black cummin, frankincense, cinnamon, cassia, myrrh, black mustard, coriander, mint, saffron, ginger grass, syrian hyssop, aloes(eagle wood), manna which have the flavor, aroma, and medical values.

동서양 종교와 철학의 기초 - '죽음'의 문제를 바라보는 두 눈 -

  • Yu, Heun-U
    • Journal of the Daesoon Academy of Sciences
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    • v.16
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    • pp.69-85
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    • 2003
  • Death has been one of the basic objects in philosophy and religion. Why do people feel afraid about death? Maybe it is because they don't know the situation after death or because they have a wrong idea about it. Someone may think that he/she doesn't care the situation. But this attitude is wrong. We must understand that our attitude about life is shaped by our knowledge of the situation after death. A recurrent theme about death in popular thought is the idea that death is mysterious. As we have seen, it is difficult to formulate a satisfactory philosophical analysis of the concept of death. If it is impossible to analyse the concept of death, then it is impossible to explain precisely what we mean when we say that something dies. It might be said therefore that, in virtue of this fact, death is mysterious. Of course, death is not distinctively mysterious - all other unanalysable concepts are equally mysterious in this way. Reflection on death gives rise to a variety of philosophical questions. One of the deepest of these is a question about the nature of death. Typically, philosophers interpret this question as a call for an analysis or definition of the concept of death. Plato, for example, proposed to define death as the separation of soul from body. However, this definition is not acceptable to those who think that there are no souls. It is also unacceptable to anyone who thinks that plants and lower animals have no souls, but can nonetheless die. Others have defined death simply as the cessation of life. This too is problematic, since an organism that goes into suspended animation ceases to live, but may not actually die. The eastern philosophers proposed to define death as the nonduality of life and death. Taoists, for example, do not believe in the Wheel of Life of the Buddhists nor in the Heaven or Hell of Christianity. Taoists view existence as glorious. The whole Universe, they teach, is a marvelous, vibrant Unity wherein everything, visible and invisible, pulses with energy and changes. As being develops through the experience of existence, its vessels are swept onwards by the mighty stream of the eternal TAO to other forms of expression and activity. Man does not die; he merely extends into new fields. Taoists teach that the end of a person is the return to the Ultimate Reality. "Life is uncertain - Death is certain": This is a well known saying in Buddhism. Knowing very well that death is certain and it is a natural phenomenon that everyone has to face, we should not be afraid of death. Yet, instinctively, all of us fear death because we do not know how to think of its inevitability. We like to cling to our life and body and so develop too much craving and attachment.

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Analysis of Nutrient Content by Digestion Phase of Legumes using an In Vitro Digestion Model (In Vitro Digestion Model을 활용한 두류 소화 단계별 영양성분 변화 분석)

  • Da Bin Lee;Kyeong A Jang;In Seon Hwang;Min Sook Kang;Mi-Kyung Seo;Haeng Ran Kim;Seon Mi Yoo
    • The Korean Journal of Food And Nutrition
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    • v.36 no.5
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    • pp.368-378
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    • 2023
  • Changes in contents of free sugars, amino acids, and fatty acids of legumes were analyzed for each phase of in vitro digestion. In addition, contents of resistant starch in raw and digested pulses were compared. Soybeans, kidney beans, cowpeas, and chickpeas were analyzed. An in vitro digestion model was used to analyze contents of nutrients using LC-MS and GC-MS. Stachyose in kidneybean, cowpea, and chickpea increased as the digestion phase progressed. In four types of legumes, raffinose slightly decreased or showed no significant difference between the Oral phase and the BBMV phase. Content of glucose, a monosaccharide, increased during the BBMV phase. During the digestion phase, levels of free amino acids and free fatty acids also increased. Content of resistant starch was reduced compared to that in the raw material. It was 0.01g/100 g food in soybean, 1.06 g/100 g food in red kidney bean, 0.77g/ 100g food in cowpea, and 0.76 g/100 g food in chickpea. It was confirmed that nutrients in the in vitro digestion model were liberated at each digestion phase with changes in the content of resistant starch. These results are expected to be used as fundamental data for obtaining bioavailability of nutrients.

Effect of Joksamni combination on NADPH-diaphorase, neuronal Nitric Oxide Synthase, Neuropeptide Y and Vasoactive Intestinal Peptide in the cerebral cortex of Spontaneously Hypertensive Rat (족삼리(足三里) 배혈(配穴)에 따른 전침(電鍼)이 흰쥐 대뇌피질(大腦皮質)의 NADPH-diaphorase와 nNOS, NPY, VIP 신경세포(神經細胞)에 미치는 영향(影響))

  • Jung, In-gy;Lee, Jae-dong;Kim, Chang-hwan
    • Journal of Acupuncture Research
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    • v.20 no.5
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    • pp.118-132
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    • 2003
  • Objective: The aim of this study was to investigate the effects of Joksamni(ST36) combination on NAD PH-diaphorase, neuronal nitric oxide synthase(nNOS), neuropeptide Y(NPY) and vasoactive intestinal peptide (VIP) in the cerebral cortex of spontaneously hypertensive rat. Methods: The experimental groups were divided into four groups: Normal, Joksamni(ST36), Joksamni(ST36)+Eumneungcheon(SP9), and Joksamni(ST36)+Gokji(LI11). Needles were inserted into acupoints at the depth of 0.5cm with basic insertion method. Electroacupuncture was done under the condition of 2Hz electrical biphasic pulses with continuous rectangular wave lasting for 0.2ms until the muscles produced visible contractions. Such stimulation was applied continuously for 10 minutes, 1 time every 2 days for 10 sessions of treatments. Thereafter we evaluated changes in NADPH-d positive neurons histochemically and changes in nNOS, NPY and VIP positive neurons immunohistochemically. Results: The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in primary somatosensory cortex, visual cortex, auditory cortex, perirhinal cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group with the exception of primary somatosensory cortex. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in auditory cortex, perirhinal cortex, insular cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. The optical densities of NPY positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in primary motor cortex, primary somatosensory cortex, cingulate cortex as compared to the Joksamni (ST36) and Joksamni(ST36)+Eumneungcheon(SP9) groups. The optical densities of VIP positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group were significantly increased in all areas of cerebral cortex except for cingulate cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in auditory cortex, cingulate cortex, perirhinal cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. Conclusions: The result demonstrated that electroacupuncture on Joksamni(ST36) and its combination change the activities of the NO system and peptidergic system in the cerebral cortex of SHR and that acupoint combination is one of the important parameters for the effects.

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${T_2}weighted$- Half courier Echo Planar Imaging

  • 김치영;김휴정;안창범
    • Investigative Magnetic Resonance Imaging
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    • v.5 no.1
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    • pp.57-65
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    • 2001
  • Purpose : $T_2$-weighted half courier Echo Planar Imaging (T2HEPI) method is proposed to reduce measurement time of existing EPI by a factor of 2. In addition, high $T_2$ contrast is obtained for clinical applications. High resolution single-shot EPI images with $T_2$ contrast are obtained with $128{\times}128$ matrix size by the proposed method. Materials and methods : In order to reduce measurement time in EPI, half courier space is measured, and rest of half courier data is obtained by conjugate symmetric filling. Thus high resolution single shot EPI image with $128{\times}128$ matrix size is obtained with 64 echoes. By the arrangement of phase encoding gradients, high $T_2$ weighted images are obtained. The acquired data in k-space are shifted if there exists residual gradient field due to eddy current along phase encoding gradient, which results in a serious problem in the reconstructed image. The residual field is estimated by the correlation coefficient between the echo signal for dc and the corresponding reference data acquired during the pre-scan. Once the residual gradient field is properly estimated, it can be removed by the adjustment of initial phase encoding gradient field between $70^{\circ}$ and $180^{\circ}$ rf pulses. Results : The suggested T2EPl is implemented in a 1.0 Tela whole body MRI system. Experiments are done with the effective echo times of 72ms and 96ms with single shot acquisitions. High resolution($128{\times}128$) volunteer head images with high $T_2$ contrast are obtained in a single scan by the proposed method. Conclusion : Using the half courier technique, higher resolution EPI images are obtained with matrix size of $128{\times}128$ in a single scan. Furthermore $T_2$ contrast is controlled by the effective echo time. Since the suggested method can be implemented by software alone (pulse sequence and corresponding tuning and reconstruction algorithms) without addition of special hardware, it can be widely used in existing MRI systems.

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Hybrid Two-Dimensional Proton Spectroscopic Imaging of Pediatric Brain: Clinical Application (소아 뇌에서의 혼성 이차원 양성자자기공명분광법의 임상적 응용)

  • Sung Won Youn;Sang Kwon Lee;Yongmin Chang;No Hyuck Park;Jong Min Lee
    • Investigative Magnetic Resonance Imaging
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    • v.6 no.1
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    • pp.64-72
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    • 2002
  • Purpose : To introduce and demonstrate the advantages of the new hybrid two-dimensional (2D) proton spectroscopic imaging (SI) over the single voxel spectroscopy (SVS) and conventional 2D SI in the clinical application of spectroscopy for pediatric cerebral disease. Materials and Methods : Eighty-one hybrid 2D proton spectroscopic imaging was performed in 79 children (36 normal infants and children, 10 with hypoxic-ischemic injury, 20 with toxic-metabolic encephalopathy, seven with brain tumor, three with meningoencephalitis, one with neurofibromatosis, one with Sturge-Weber syndrome and one with lissencephaly) ranging in age from the third day of life to 15 years. In adult volunteers (n=5), all three techniques including hybrid 2D proton SI, SVS using PRESS sequence, and conventional 2D proton SI were performed. Both hybrid 2D proton SI and SVS using PRESS sequence were performed in clinical cases (n=). All measurements were performed with a 1.5-T scanner using standard head quadrature coil. The 16$\times$16 phase encoding steps were set on variable field of view (FOV) depending on the size of the brain. The hybrid volume of interest inside FOV was set as $75{\times}75{\times}15{\;}\textrm{mm}^3$ or smaller to get rid of unwanted fat signal. Point-resolved spectroscopy (TR/TE=1,500 msec/135 or 270msec) was employed with standard chemical shift selective saturation (CHESSI pulses for water suppression. The acquisition time and spectral quality of hybrid 2D proton SI were compared with those of SVS and conventional 2D proton SI. Results : The hybrid 2D proton SI was successfully conducted upon all patients.

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Studies on production of nuclear transplanted mouse embryos (핵이식 마우스 생산에 관한 연구)

  • Lee, Byeong-chun;Jo, Choong-ho;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.151-169
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    • 1993
  • The present study was carried out to investigate the best condition for nuclear-cytoplasm fusion and in vitro culture of nuclear transplanted embryos and to investigate the production of nuclear transplanted offsprings. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulses at output voltage of 1.0, 1.5 and 2.0 kV/cm for 100, 150 and $200{\mu}$ sec to induce cell fusion. 1. The culture of intact or zona cut 2-cell embryos in the medium supplemented with cytochalasin B($5{\mu}g/m{\ell}$) and colcemide($0.1{\mu}g/m{\ell}$)for 30 and 60 minutes did not affect the development to later stage. 2. The in vitro developmental rates of group A(a nucleus from one of the blastomeres was removed) and B(electrofusion of group A) were significantly lower than that of control group(p<0.01). 3. When nuclear transplanted embryos were submitted to electrofusion, the significantly higher fusion rates of 2-cell donor nuclei were achieved at the electric field strength of DC 1.5kV/cm for 100 and $150{\mu}$ sec, DC 2.0 kV/cm for $100{\sim}200{\mu}$ sec than DC 1.0 kV/cm for 100 and $150{\mu}$ sec(p<0.01). The significantly higher fusion rates of 4-cell donor nuclei were achieved at DC 2.0 kV/cm for 100 and $150{\mu}$ sec than DC 1.0kV/cm for $100{\sim}200{\mu}$ sec(p<0.01). These fusion rates in 8-cell donor nuclei were 88.7~99.3%. 4. The developmental potency to blastocyst in 2- and 4-cell donor nuclei was significantly higher in DC 1.0 and 2.0 kV/cm for $100{\sim}200{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group (p<0.01). The developmental potency to blastocyst in 8-cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}$ sec treated group than in DC 1.0 kV/cm for $100{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group(p<001). 5. The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 6. The success rate of nuclear injection into enucleated 2-cell embryos was significantly higher in 2-cell donor nuclei than in 4- or 8-cell donor nuclei(p<0.01). 7. The culture time taken for the nuclear transplanted 2-cell embryos to blastocyst stage was significantly longer in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 8. There was no significant difference in the developmental potency of nuclear transplanted embryos within the concentration of EGF at 0 to 15 ng per $m{\ell}$ of BMOC-3 solution. 9. The production rates of offspring after transfer of nuclear transplanted embryos to recipient mouse were significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01).

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Effect of $K^+-channel$ Blockers on the $A_1-adenosine$ Receptor-Coupled Regulation of Electrically-Evoked Norepinephrine Release in the Rat Hippocampus (흰쥐 해마에서 Norepinephrine 유리를 조절하는 $A_1-adenosine$ 수용체의 역할에 미치는 $K^+$ 통로 차단제의 영향)

  • Choi, Bong-Kyu;Kim, Sang-Hoon
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.301-309
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    • 1996
  • Since it has been reported that the depolarization-induced NE release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the NE release has not been clearly elucidated yet. Therefore, it was attempted to clarify the participation of $K^+-channel$ in the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of NE release in this study. Slices from rat hippocampus were equilibrated with $^3H-norepinephrine$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $VCm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $1{\sim}30\;{\mu}M$, decreased the NE release in a dose-dependent manner, without affecting the basal rate of release. 4AP $(1{\sim}30{\mu}M)$, a specific A-type $K^+-channel$ blocker, increased the evoked NE release in a dose-related fashion, and the basal rate of release is increased by 10 and $30{\mu}M$. TEA $(1{\sim}10{\mu}\;M)$, a nonspecific $K^+-channel$ blocker, increased the evoked NE release in a dose-dependent manner without affecting basal release. The adenosine effects were significantly inhibites by 3 ${\mu}M$ 4AP and 10 mM TEA treatment. 4AP $(30{\mu}M)-$ and TEA (10 mM)-induced increments of evoked NE release were completely abolished in $Ca^{++} free, but these were recoverd in low $Ca^{++} medium. And the effects of $K^+-channel$ blockers in low $Ca^{++} medium were inhibites and abolishes by $Mg^{++} (4 mM) adding and TTX $(0.3{\mu}M)$ adding medium, respectively. These results suggest that the decrement of the evoked NE-release by $A_1-adenosine$ receptor is mediated by 4AP and TEA sensitive $K^+-channel$.

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The Role of Adenosine Receptors on Acetylcholine Release in the Rat Striatum

  • Kim, Do-Kyung;Kim, Hyeon-A;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.1-12
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    • 1997
  • As it has been reported that the depolarization induced acetylcholine (ACh) release is modulated by activation of presynaptic $A_1$ adenosine heteroreceptor and various evidence suggest that indicate the $A_2$ adenosine receptor is present in the striatum, this study was undertaken to delineate the role of adenosine receptors on the striatal ACh release. Slices from the rat striatum were equilibrated with $[^3H]$choline and then the release amount of the labelled product, $[^3H]$ACh, which was evoked by electrical stimulation (rectangular pulses, 3 Hz, 2 ms, 24 mA, $5\;Vcm^{-1}$, 2 min), was measured, and the influence of various agents on the evoked tritium outflow was investigated. And also, quantitative receptor autoradiography and drug-receptor binding assay were performed in order to confirm the presence and characteristics of $A_1$ and $A_2$ adenosine receptors in the rat striatum. Adenosine $(10{sim}100\;{mu}M)$ and $N^6$-cyclopentyladenosine (CPA, $1{sim}100\;{mu}M)$ decreased the $[^3H]$ACh release in a dose-dependent manner without changing the basal rate of release in the rat striatum. The reducing effects of ACh release by adenosine and CPA were abolished by 8-cyclopentyl-1,3-dipropy-Ixanthine (DPCPX, 2 ${mu}M$), a selective $A_1$, adenosine receptor antagonist, treatment. The effect of adenosine was potentiated markedly by 3,7-dimethyl-1-propargylxanthine (DMPX, 10 ${mu}M$), a specific $A_2$ adenosine receptor antagonist. 2-P-(2-carboxyethyl)phenethylamimo-5'-N- ethylcarboxamidoadenosine hydrochloride (CGS-21680C), in concentrations ranging from 0.01 to 10 ${mu}M$, a recently introduced potent $A_2$ adenosine receptor agonist, increased the $[^3H]$ACh release in a dose related fashion without changing the basal rate of release. These effects were completely abolished by DMPX $(10\;{mu}M)$. In autoradiograrhy experiments, $[^3H]$2-chloro-$N^6$-cyclopentyladenosine ($[^3H]$ CCPA) bindings were highly localized in the hippocampus and the cerebral cortex. Additionally, lower levels of binding were found in the striatum. However, $[^3H]$CGS-21680C bindings were highly localized in the striatal region with the greatest density of binding found in the caudate nucleus and putamen. Lower levels of binding were also found in the nucleus accumbens and olfactory tubercle. In drug-receptor binding assay, binding of $[^3H]$ CCPA to $A_1$ adenosine receptors of rat striatal membranes was inhibited by CPA ($K_i$ = 1.6 nM) and N-ethylcarboxamidoadenosine (NECA, $K_i$ = 12.9 nM), but not by CGS-21680C ($K_i$ = 2609.2 nM) and DMPX ($K_i$ = 19,386 nM). In contrast, $[^3H]$CGS-21680C binding to $A_2$ denosine receptors was inhibited by CGS-21680C ($K_i$ = 47.6 nM) and NECA ($K_i$ = 44.9 nM), but not by CPA ($K_i$ = 2099.2 nM) and DPCPX ($K_i$ = 19,207 nM). The results presented here suggest that both types of $A_1$ and $A_2$ adenosine heteroreceptors exist and play an important role in ACh release in the rat striatal cholinergic neurons.

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