• 대한생식의학회:학술대회논문집
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• 대한불임학회 1999년도 제38차 추계 학술대회
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• pp.105-106
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• 1999

• The Korean Journal of Physiology and Pharmacology
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• 제3권3호
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• pp.339-350
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• 1999

The present study was attempted to examine the effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on catecholamine (CA) secretion evoked by cholinergic stimulation, membrane depolarization and calcium mobilization from the isolated perfused rat adrenal gland. The perfusion of PACAP (10 nM) into an adrenal vein for 60 min produced a great inhibition in CA secretion evoked by ACh $(5.32{\times}10^{-3}\;M),$ high $K^+\;(5.6{\times}10^{-2}\;M),$ DMPP $(10^{-4}\;M\;for\;2\;min),$ McN-A-343 $(10^{-4}\;M\;for\;2\;min),$ cyclopiazonic acid $(10^{-5}\;M\;for\;4\;min)$ and Bay-K-8644 $(10^{-5}\;M\;for\;4\;min).$ Also, in the presence of neuropeptide (NPY), which is known to be co-localized with norepinephrine in peripheral sympathetic nerves, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were also significantly depressed. However, in adrenal glands preloaded with PACAP (10 nM) under the presence of VIP antagonist $[(Lys^1,\;Pro^{2.5},\;Arg^{3.4},\;Tyr^6)-VIP\;(3\;{\mu}M)]$ for 20 min, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were not altered greatly in comparison to the case of PACAP-treatment only. Taken together, these results suggest that PACAP causes the marked inhibition of CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization, indicating that this effect may be mediated by inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenomedullary chromaffin cells.

• Asian-Australasian Journal of Animal Sciences
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• 제32권12호
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• pp.1844-1853
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• 2019

Objective: We investigated how pituitary adenylate cyclase-activating polypeptide (PACAP) affects embryonic development during pre-in vitro maturation (pre-IVM) using porcine oocytes isolated from small follicles. Methods: We divided the follicles into the experimental groups by size (SF, small follicles; MF, medium follicles) and treated with and without PACAP and cultured for 18 hours (PreSF[-]PACAP; without PACAP, Pre-SF[+]PACAP; with PACAP) before undergoing IVM. The gene expression related to extracellular matrix formation (amphiregulin, epiregulin, and hyaluronan synthase 2 [HAS2]) and apoptosis (Bcl-2-associated X [BAX], B-cell lymphoma 2, and cysteine-aspartic acid protease 3) was investigated after maturation. The impact on developmental competence was assessed by the cleavage and blastocyst rate and total cell number of blastocysts in embryos generated from parthenogenesis (PA) and in vitro fertilization (IVF). Results: Cleavage rates in the Pre-SF(+)PACAP after PA were significantly higher than SF and Pre-SF(-)PACAP (p<0.05). The cleavage rates between MF and Pre- SF(+)PACAP groups yielded no notable differences after IVF. Pre-SF(+)PACAP displayed the higher rate of blastocyst formation and greater total cell number than SF and Pre-SF(-)PACAP (p<0.05). Cumulus cells showed significant upregulation of HAS2 mRNA in the Pre-SF(+)PACAP compared to the SF (p<0.05). In comparison to other groups, the Pre-SF(+)PACAP group displayed a downregulation in mRNA expression of BAX in matured oocytes (p<0.05). Conclusion: The PACAP treatment during pre-IVM improved the developmental potential of porcine oocytes derived from SF by regulating cumulus expansion and apoptosis of oocytes.

• 한국미생물·생명공학회지
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• 제38권4호
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• pp.349-361
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• 2010

본 총설에서는 아미노산의 공업적 생산균인 Corynebacterium glutamicum의 탄소 대사 및 이와 관련된 총체적 조절 메커니즘에 대한 최근의 연구를 정리하였다. C. glutamicum의 산업적 발효을 위한 기질로서 사용되는 당밀은 주로 sucrose, glucose, fructose로 이루어져 있으며, 이들 당은 phosphotransferase system을 통해서 수송된다. C. glutamicum의 탄소 대사 특징은 glucose가 다른 당이나 유기산 등과 함께 존재할 때, glucose와 이러한 탄소원 들을 동시에 대사한다. 그러나 glucose/glutamate 혹은 glucose/ethanol 등의 혼합물에서 는 탄소원의 순차적 이용으로 인해 나타나는 diauxic growth 현상을 나타내며, 이러한 carbon catabolite repression(CCR) 현상은 E. coli나 B. subtilis 등에서 알려진 것과는 다른 독특한 분자적 메커니즘과 조절 circuits을 가지고 있음이 밝혀지고 있다. C. glutamicum의 CRP homologue인 GlxR은 acetate 대사를 포함하여 glycolysis, gluconeogenesis 및 TCA cycle 등을 포함하는 중심탄소대사 조절 뿐만 아니라, 다양한 세포 기능의 조절에 관여하는 총체적 조절 단백질로서의 역할이 제시되고 있다. C. glutamicum의 adenylate cyclase(AC)는 막과 결합된 class IIIAC 로서, 막 단백질의 특성상 아직 규명되어 있지 않은 세포 외부의 환경 변화에 대응하여 세포 내의 cAMP합성 수준을 조절할 수 있는 sensor로 추정할 수 있다. 특히 C. glutamicum의 경우 배지내 glucose 를 비롯한 탄소원과 cAMP 농도와의 관련성이 E. coli에서 알려진 교과서적 지식과는 상반되게 변화하는 경향을 보이고 있어, cAMP signaling에 의한 세포 내 regulatory network 등은 향후 풀어야 할 의문으로 남아있다. 탄소대사 조절의 최상위에 존재하며 global 조절자인 GlxRcAMP 복합체 이외에도 차상위 전사조절 단백질로서 RamB, RamA, SugR 등이 존재하여 다양한 탄소대사를 조절한다. 최근 들어서는 새로운 탄소원으로서 대두되고 있는 biomass 관련 기질들을 이용할 수 있는 C. glutamucum 균주 구축을 통하여 이용 기질의 범위를 확대시키고자 하는 연구 및 탄소 대사와 관련하여 L-lysine의 발효 수율 혹은 생산성을 향상시키고자 하는 다양한 분자적 균주 육종 연구 등이 수행되고 있다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1984년도 학술대회지
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• pp.107-111
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• 1984

쥐의 뇌에서 추출한 입자상 아데닐산 고리화효소와 입자상 구아닐산효소의 활동성에 미치는 몇 가지 긴세노시드들의 효과를 조사하였다. $Rb_{2},\;Rb_{1}$, Rc 및 Re들과 같은 약간의 진세노사이드들이 두 효소들의 활동성을 상반적으로 변화시키는 것을 관찰하였다. 아데닐산 고리화효소와 구아닐산 고리화효소의 활동성에 미치는 GMP 및 AMP의 조절작용을 조사하였다. 긴세노시드 Rd로 방해된 아데닐산고리화효소는 GMP를 첨가함에 따라서 활성화되었다. 마찬가지로. 긴세노시드 $Rb_{2}$로 방해된 아데닐산 고리화효소도 GMP에 의해서 활성화되었다. 다른 한편, 긴세노시드 Rc로 활성화된 구아닐산 고리화효소는 AMP 또는 GMP를 첨가함에 따라서 방해되었다. 진세노사이드 $Rb_{2}$ 도파민 사이에는 아데닐산 고리화효소 계상의 수용체들에의 결함에 있어서 경쟁적이라는 것을 알았다. 이 결과는 긴세노시드 $Rb_{2}$가 세포의 D-1 도파민 수용체에 특이하게 결합한다는 것을 말해 준다.

• The Korean Journal of Physiology and Pharmacology
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• 제25권3호
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• pp.227-237
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• 2021

Carbon monoxide (CO) is a cardioprotectant and potential cardiovascular therapeutic agent. Human cardiac fibroblasts (HCFs) are important determinants of myocardial structure and function. Large-conductance Ca2+-activated K+ (BK) channel is a potential therapeutic target for cardiovascular disease. We investigated whether CO modulates BK channels and the signaling pathways in HCFs using whole-cell mode patch-clamp recordings. CO-releasing molecules (CORMs; CORM-2 and CORM-3) significantly increased the amplitudes of BK currents (IBK). The CO-induced stimulating effects on IBK were blocked by pre-treatment with specific nitric oxide synthase (NOS) blockers (L-NG-monomethyl arginine citrate and L-NG-nitroarginine methyl ester). 8-bromo-cyclic GMP increased IBK. KT5823 (inhibits PKG) or ODQ (inhibits soluble guanylate cyclase) blocked the CO-stimulating effect on IBK. Moreover, 8-bromo-cyclic AMP also increased IBK, and pre-treatment with KT5720 (inhibits PKA) or SQ22536 (inhibits adenylate cyclase) blocked the CO effect. Pre-treatment with N-ethylmaleimide (a thiol-alkylating reagent) also blocked the CO effect on IBK, and DL-dithiothreitol (a reducing agent) reversed the CO effect. These data suggest that CO activates IBK through NO via the NOS and through the PKG, PKA, and S-nitrosylation pathways.

• Molecules and Cells
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• 제26권2호
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• pp.181-185
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• 2008

The effects of calcitonin gene-related peptide (CGRP) on pacemaker currents in cultured interstitial cells of Cajal (ICC) from the mouse small intestine were investigated using the whole-cell patch clamp technique at $30^{\circ}C$. Under voltage clamping at a holding potential of -70 mV, CGRP decreased the amplitude and frequency of pacemaker currents and activated outward resting currents. These effects were blocked by intracellular $GDP{\beta}S$, a G-protein inhibitor and glibenclamide, a specific ATP-sensitive $K^+$ channels blocker. During current clamping, CGRP hyperpolarized the membrane and this effect was antagonized by glibenclamide. Pretreatment with SQ-22536 (an adenylate cyclase inhibitor) or naproxen (a cyclooxygenase inhibitor) did not block the CGRP-induced effects, whereas pretreatment with ODQ (a guanylate cyclase inhibitor) or L-NAME (an inhibitor of nitric oxide synthase) did. In conclusion, CGRP inhibits pacemaker currents in ICC by generating nitric oxide via G-protein activation and so activating ATP-sensitive $K^+$ channels. Nitric oxide- and guanylate cyclase-dependent pathways are involved in these effects.

• 대한수의학회지
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• 제22권1호
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• pp.1-8
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• 1982

The effect of acetylcholine, oxytocin and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) on cyclic nucleotide levels in estrogen-primed rabbit whole uterus were studied in the presence and absence of 1-methyl-3-isobutyl xanthine (MIX), a phosphodiestrase inhibitor, and indomethacin, a prostagandin inhibitor. In the absence of MIX, acetylcholine increased guanosine 3', 5'-cyclic monophosphate (cGMP), but had no effect on adenosine 3', 5'-cyclic monophosphate (cAMP) levels. In contrast, oxytocin had no influence on cGMP, but decreased cAMP levels. $PGF_{2{\alpha}}$ increased cGMP and decreased cAMP levels. MIX increased both cAMP and cGMP levels. Oxytocin and $PGF_{2{\alpha}}$ further increased cGMP levels, indicating activation of guanylate cyclase activity. The ratio of cAMP/cGMP was decreased by uterine stinulants both in presence and absence of MIX. Indomethacin elevated cAMP and cGMP revels. The effects of uterine stimulants in the presence of indomethacin on cyclic nucleotide levels were varied from tissue to tisse. In general, oxytocin decreased cGMP and $PGF_{2{\alpha}}$ increased cAMP/cGMP levels, but the effects were statisically nonsignicficant. The cAMP/cGMP ratio was increased by uterine stimulant in the presence of indomethacin. In conclusion, uterine stimulants eased cAMP/cGMP ratio which indicates that the uterine stimulants have opposing effects on adenylate cyclase and guanylate cyclase activities. The endometrium plays a role in the regulation of cyclic nucleotide levels and uterine contraction by means of PG synthesis. Indomethacin has an unknown activities besides both of PG synthetase and phosphodiesterase inhibitions.

• Journal of Ginseng Research
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• 제21권3호
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• pp.141-146
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• 1997

The role of cAMP in the differentiation process of F9 cells induced by ginsenosides was examined by performing transient transfixion assay with CRE-luciferase reporter plasmid, GR thansactivation assay with GRE-luciferase activity with or without treatment of CAMP and forskolin, an activator of adenylate cyclase, and protein klnase A assay in the presence of ginsenosides. Ginsenosides had no effect on CRE-transactivation activity, whereas retinoic acid induced the activity. When cAMP or forskolin was treated with ginsenosides, GRE-luciferase activity was further augumented by them. In addition, ginsenosides induced protein kinase A activity in the presence of cAMP. These results suggest that ginsenosides activate cAMP-dependent protein kinase A which, in turn, increase GR activity in F9 cells.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.292-292
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• 1994

Catecholamines acting through ${\beta}$-adrenergic receptors regulate a wide range of metabolic activities in mammalian tissue. Of the various receptors coupled to adenylate cyclase, the ${\beta}$-adrenergic receptors are the most extensively characterized and have been purified from both nonmammal ian and mammal inn sources. However, most studies of the molecular properties of ${\beta}$-adrenergic receptors have been confined to peripheral tissues. Less progress has been achieved in characterizing the brain ${\beta}$-adrenergic receptor The goal of the present study was, therefore, to purify and characterize the neurotransmitter receptor proteins. To achieve this goal, the following stepwise experiments were performed. At first, the membrane-bound ${\beta}$-adrenergic receptors were. solubi1ized from brain tissue. Secondly, conditions for affinity chromatography were determined to purify the solubilized receptors effectively. Finally, the large-scale purification was performed and the characteristics of the purified ${\beta}$-adrenergic receptor were examined.