• 대한수의학회지
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• 제31권4호
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• pp.389-395
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• 1991

가토 적출 신동맥에 있어서 purinergic 신경 분포를 perivascular nerve stimulation을 통해서 확인하였다. 1. 가토 적출 신동맥은 perivascular nerve stimulation에 의해 수축성 반응을 나타내었다. 2. Perivascular nerve stimulation에 의한 수축현상은 자극의 크기와 자극의 빈도에 의존적이었으며, 내피세포의 유무에는 영향을 받지 않았다. 3. Perivascular nerve stimulation에 의한 수축현상은 prazosin$(1{\mu}M)$에 의해 강하게 억제되어졌으며, tetrodotoxin$(1{\mu}M)$ 또는 ${\alpha},{\beta}-methylene$ATP$(10{\mu}M)$에 의한 $P_{2x}-purinoceptor $ desensitization에 의해 완전히 수축현상이 사라졌다. 이와같은 결과는 가토 신동맥에는 purinergic신경이 분포되어 있으며, perivascular nerve stimulation에 의해 purinergic 신경전달물질이 norepinephrine과 함께 유리되어지는 것을 시사하고 있다.

• 대한약리학회지
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• 제30권2호
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• pp.181-190
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• 1994

In rat atrium the characteristics of purinergic receptors were investigated by observing the effects of some purinergic receptor agonists and antagonists on action potential and contractile force. The statistically significant effects of $ATP(10^{-6}{\sim}10^{-3}M)$ and adenosine $(10^{-6}{\sim}10^{-3}M)$ on normal action potential characteristics were a dose-dependent shortening of action potential duration $(APD_{90})$ by both agents and hyperpolarization by $ATP(10^{-4},10^{-3}M)$. $CAP(10^{-8}{\sim}10^{-4}M)$, an $A_1$ adenosine receptor agonist, shortened $(APD_{90})$ markedly in a dose-dependent manner and these effects were almost abolished by $DPCPX\;(10^{-6}\;M), an $A_1$, adenosine receptor antagonist, but not affected by $DMPX(2{\times}10^{-6}\;M)$, an $A_2$ adenosine receptor agonist. On the other hand, CGS $21680(10^{-7}{\sim}10^{-4}M)$, an $A_2$ adenosine receptor agonist, elicited a slight shortening of $(APD_{90})$ and these effects were inhibited by DPCPX but persisted in the presence of DPMX. Adenosine $(10^{-6}{\sim}10{\-4}\;M)$ decreased the basal contraction of atrial muscle in a dose-dependent manner and these effects were not inhibited by DMPX but by DPCPX. These results suggests that purinergic receptor agonists depress the cardiac activity by a short ening of action potential duration and this effect is mostly mediated by $A_1$ adenosine receptors in rat atrium.

• 대한의생명과학회지
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• 제13권4호
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• pp.333-339
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• 2007

Recently, much attention has been paid to human retinoblastoma since it provide a good model system for studying mechanisms underlying cell growth, differentiation, proliferation, and apoptosis, and for developing cancer therapy. However, until now it is unclear whether purinergic receptors are involved in the calcium mobilization in the retinoblastoma cells. In this regard, we measured possible purinergic signaling in WERI-Rb-1 cells using $Ca^{2+}$ imaging technique and RT-PCR method. ATP-induced $[Ca^{2+}]_i$ transients was maintained to about $90.7{\pm}1.0%$ of the control (n=48) even in the absence of extracellular calcium. The ATP-induced intracellular calcium response was only attained to $10.4{\pm}1.8%$ (n=55) of peak amplitude of the control after preincubation of 1 ${\mu}MU-73122$, a PLC inhibitor, but it was not affected by 1 ${\mu}MU-73343$, a inactive form of U-73122. And also ATP-induced $[Ca^{2+}]_i$ rise was almost attenuated by 20 ${\mu}M$ 2-APB, a putative $IP_3$ receptor inhibitor. Two subtypes of $IP_3$ receptor $(IP_{3-1}R,\;IP_{3-2}R)$ were identified by a RT-PCR method. These findings suggest that purinergic stimuli can cause calcium mobilization via $PLC-IP_3$ pathway after the activation of P2Y receptors in the retinoblastoma cells, which may play important roles in cell proliferation, differentiation, growth, and cell death.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1997년도 학술발표회
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• pp.16-16
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• 1997

Extracelluar ATP evokes many biological processes, including neuronal excitation and neurotransmitter secretion, through activation of purinergic P2 receptors. Although excitatory and inhibitory receptor-operated channels (ROC) and voltage-dependent calcium channels (VDCC) have been reported to be altered by acute and chronic exposure to ethanol, little is known of the ethanol effects on purinergic receptor-operated channels in neuronal cells.(omitted)

• BMB Reports
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• 제51권9호
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• pp.468-473
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• 2018

Purinergic receptor signaling is increasingly recognized as an important regulator of inflammation. The P2X family purinergic receptors P2X5 and P2X7 have both been implicated in bone biology, and it has been suggested recently that P2X5 may be a significant regulator of inflammatory bone loss. However, a role for P2X5 in periodontitis is unknown. The present study aimed to evaluate the functional role of P2X5 in ligature-induced periodontitis in mice. Five days after placement of ligature, analysis of alveolar bone revealed decreased bone loss in $P2rx5^{-/-}$ mice compared to $P2rx7^{-/-}$ and WT control mice. Gene expression analysis of the gingival tissue of ligated mice showed that IL1b, IL6, IL17a and Tnfsf11 expression levels were significantly reduced in $P2rx5^{-/-}$ compared to WT mice. These results suggest the P2X5 receptor may regulate bone loss related to periodontitis and it may thus be a novel therapeutic target in this oral disease.

• 대한수의학회지
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• 제37권3호
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• pp.533-540
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• 1997

To elucidate the effects of purinergic nerve on relaxation of pig coronary artery, the effects of ATP, 2-methylthio ATP and electrical perivascular nerve stimulation were investigated from physiograph on the isolated coronary artery of pig. The results btained were as follows; 1. The relaxative responses induced by perivascular nerve stimulation(20V, 0.5msec, 10sec) were the frequency(1~8Hz) dependent manner with phentolamine($10^{-5}M$) and atropine($10^{-6}M$) on isolated coronary artery of pig. 2. The relaxative responses induced. by adenosine($10^{-7}{\sim}5{\times}10^{-3}M$) or ATP($10^{-7}{\sim}5{\times}10^{-5}M$) on precontraction with histamine($10^{-5}M$) were the dose-dependent manner, but the contractile responses were often induced by ATP($10^{-4}M$ and $10^{-3}M$). 3. The relaxative responses induced by 2-methylthio ATP($2.5{\times}10^{-8}{\sim}2.5{\times}10^{-6}M$) on precontraction with histamine($10^{-5}M$) were the dose-dependent manner. 4. The relaxative response induced by 2-methylthio ATP($10^{-7}M$) on precontraction with histamine($10^{-5}M$) was completely blocked by the pretreatment with $P_{2Y}$-purinoceptor blocker, reactive blue 2($10^{-4}M$). 5. The neurogenic relaxative response induced by perivascular nerve stimulation(20V, 8Hz, 0.5msec, 10sec) was weakly inhibited by the pretreatment with ${\beta}$-adrenoceptor blocker, propranolol($10^{-5}M$) and blocked by the addition with $P_{2Y}$-purinoceptor blocker, reactive blue 2($10^{-4}M$). The results suggest that the purinergic nerve is innervated, and its relaxative response was mediated by $P_{2Y}$-purinoceptor on isolated coronary artery in pig.

• Journal of Yeungnam Medical Science
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• 제8권2호
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• pp.138-149
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• 1991

흰쥐(Sprague-Dawley)의 방광에서 적출한 배뇨근절편을 적출 근편 실험조에 현수하고, 등척성 장력 측정기를 사용하여 그 수축력을 묘기하였다. 배뇨근 절편은 전기장자극에 의해 수축하였으며, 이 수축반응은 콜린에스테라제 억제약물인 physostigmine에 의해 증가하였고 신경말단에서의 choline 재흡수를 방해하는 hemicholinium에 의해 억제되었으며, 신경절봉쇄약물인 hexamethonium에 의해서는 영향을 받지 않았으나 신경축색전도 억제제인 tetrodotoxin에 의해서 소실되었다. 이러한 전기장자극유발 수축은 콜린성 무스카린성 수용체봉쇄약물인 atropine에 의해 부분적으로 길항되었으며, atropine에 의해 길항되지 않는 부분은 ATP 탈감작에 의해 완전히 소실되었다. 배뇨근 절편은 콜린성 무스카린성 수용체 흥분제인 bethanechol과 퓨린성 수용체 흥분제인 ATP에 의하여 농도의존적 수축력 증가를 나타내었으며, 이중 bethanechol 유발수축은 ATP 탈감작에 의해 영향을 받지 않았고, ATP 유발수축은 tetrodotoxin에 의하여 영향을 받지 않았다. 이상의 결과로 보아 흰쥐의 적출배뇨근에는 흥분성 신경전달체계로서 퓨린성 수용체와 콜린성 수용체가 존재하며, 이들은 서로 영향을 미침이 없이 독자적으로 배뇨근 수축에 기여하고 있다고 사료된다.

• 대한수의학회지
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• 제38권4호
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• pp.730-736
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• 1998

To elucidate whether or not innervation of purinergic nerve and functional action of $P_{2X}$-, $P_{2Y}$-purinoceptor, the neurogenic effects of perivascular nerve stimulation were investigated using by physiograph recording system in isolated coronary artery of pig. 1. The contractile responses induced by perivascular nerve stimulation (20V, 0.5msec, 20sec) were the frequency (2~64Hz) dependent manner. 2. The neurogenic contractile responses induced by perivascular nerve stimulation were inhibited by the pretreatment with either ATP or adenosine ($10^{-7}{\sim}10^{-4}M$). 3. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were increased by the pretreatment with reactive blue 2, but were not affected by the pretreatment with 8-phenyltheopylline ($10^{-5}M$). 4. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were inhibited by the desensitization of the P2X-purinoceptor using by treatment of $10^{-5}M$, -methylene ATP as 3 times over again. The accomplished present study on isolated coronary artery of pig suggest that purinergic nerve is innervated and that the neurogenic contractile response was mediated by activation of $P_{2X}$-purinoceptor and the neurogenic relaxative response was mediated by activation of both $P_1$ and $P_{2Y}$-purinoceptor.

• 미생물학회지
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• 제47권1호
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• pp.7-13
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• 2011

일반적으로 대장균을 숙주로 이용하여 고등생물 유래 막단백질을 발현시킬 경우 발현된 막단백질은 숙주 세포에 치명적인 독성을 보인다. 우리가 발현을 시도한 15개의 인간 막단백질 중에서 특히 퓨린수용체 $P2X_4$ 발현은 대장균에 강한 독성을 보였다. 이러한 독성의 원인을 알아보기 위해 hydroxylamine을 사용하여 하여 인간 $P2X_4$ 유전자를 돌연변이 시키고 독성이 약해진 돌연변이체를 선별하였다. 돌연변이체 단백질을 면역블랏으로 분석한 결과 야생형에 비해 모두 단백질의 크기가 작았다. 크기가 제법 큰 돌연변이 두 개를 골라 DNA 서열분석을 해보니 130번째, 또는 194번째 Trp 코돈이 종결코돈으로 바뀜으로써 두 번째 막통과 도메인이 사라진 truncated protein이라는 사실을 알았다. 이들 돌연변이체의 세포내 위치를 추적해보니 둘 다 세포막에 삽입되어 있지는 않았다. 이런 결과를 종합해 볼 때 $P2X_4$의 발현이 대장균에 독성을 보이기 위해서는 전체 단백질의 올바른 세포막 삽입이 중요함을 시사한다.

• 대한의생명과학회지
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• 제15권4호
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• pp.319-326
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• 2009

Extracellular ATP elicits diverse physiological effects by binding to the G-protein-coupled P2Y receptors on the plasma membrane. In addition to the short-term effects of extracellular nucleotides on cell functions, there is evidence that such purinergic signalling can have long-term effects on cell proliferation, differentiation and death. The 3T3-L1 cell line derived from mouse embryo is a well-established and commonly utilized in vitro model for adipocytes differentiation and function. However, the distributions and roles of P2Y subtypes are still unknown in the preadipocyte. In this study, we identified the distributions and roles of P2Y subtypes in preadipocyte using $Ca^{2+}$ imaging and realtime PCR. ATP increased the $[Ca^{2+}]_i$ in a concentration-dependent manner. ATP increased $Ca^{2+}$ in absence and/or presence of extracellular $Ca^{2+}$. Suramin, non-selective P2Y blocker, largely blocked the ATP-induced $Ca^{2+}$ response. U73122, a PLC inhibitor, completely inhibited $Ca^{2+}$ mobilization in 3T3-L1 cells. The mRNA expression by realtime PCR of P2Y subtypes was $P2Y_2:P2Y_5:P2Y_6=1.0:12.5:0.3$. In conclusion, we showed that $P2Y_5$ receptor is a dominant purinergic receptor in preadipocytes, and multiple P2Y receptors could involve in differentiation and migration via regulating of intracellular calcium concentration.