• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.885-888
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• 2000

Electroantennogram(EAG) can be applied to detect odorants since insects have highly specialized olfactory receptors inside their antennae. The characteristics of EAG parameters were investigated for the quantitative measurement of a general odorant using ammonia as a model odorant. The antennae of male silkworm moth, Bombyx mori, were used for the EAG. The electical signal curves generated from a pair of antennae originating from the same silkworm moth were never identical; however, they exhibited a typical type I or type II characteristic curve pattern for every pair of antennae. The correlation between the EAG parameters and the ammonia concentration was analyzed for the type I and type II antennae. The stability of each parameter was also investigated for each type of antenna. The results show the possibility of the quantitative measurement of general odorants using the EAG technique.

• Reproductive and Developmental Biology
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• 제31권1호
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• pp.15-20
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• 2007

가축의 번식과 수요를 조절할 수 있는 생물학적 자극 통제 수단으로 새로운 돼지 페르몬성 냄새 물질를 탐색하고자, 기질 분자로서 2-cyclohexyloxytetrahydrofurane (A), 2-phenoxytetrahydrofurane (B) 유도체들의 설명인자인 물리화학 파라미터와 돼지 페로몬의 수용체 (pOBP)에 대한 결합 친화력 상수($p[Od.]_{50}$) 간의 2D-QSAR 모델을 유도하고 검토하였다. 2D-QSAR 모델은 결합 친화력 상수를 약 96.4% 설명하는 매우 양호한 모델($r^{2}=0.964$)로서 분자내 치환기의 소수성 (SL) 상수에 관한 적정값이 $(SL)_{opt.}=1.418$일 때 가장 높은 결합 친화력을 나타냄을 알았다. 그러므로 분자내 치환기의 소수성 인자가 결합 친화력 상수에 가장 큰 영향을 미치는 중요한 요소이었다.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.117-122
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• 2010

To search the potent pig pheromonal odorants through receptor-based approach methods, molecular dockings between 680 Flavomets as substrate molecule and pig odorants binding proteins OBP (1HQP) and PBP (1GM6) as receptor, and QSPR (quantitative structure-property relationship) analyses from physico-chemical parameters of Flavomets and their docking scores (DS) were performed and discussed quantitatively. From the basis on the findings, the optimal value $(MSA)_{opt.}=407.595\;{\AA}^2$ of MSA (molecular surface area; ${\AA}$), and RB (number of rotational bond) had the Flavomets will be able to increase DS. Therefore, it is expected that the stearyl alcohol from DS and H-bond type between substrate and receptor would be shows the character as potent pig pheromonal odorant.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.928-933
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• 2007

Cameleon is a genetically engineered $Ca^{2+}$ sensing molecule consisting of two variants of the green fluorescent protein (GFP), calmodulin and calmodulin-binding protein, M13. HEK-293 cells stably expressing three types of cameleons, yellow cameleon-2, cameleon-3er, and cameleon-2nu, were constructed, and the expression and localization of these cameleons were confirmed by fluorescent imaging. Among the cameleons, the yellow cameleon-2 was selected for analyzing the change in $Ca^{2+}$ induced by the olfactory receptor-mediated signal transduction, because it is localized in the cytosol and binds to cytosolic $Ca^{2+}$ ions. Cells stably expressing yellow cameleon-2 were transfected with each of the test olfactory receptor genes, odr-10 and 17, and the expression of the olfactory receptor genes were examined using immunocytochenmical methods and RT-PCR. Stimulating each olfactory receptor with its specific odorant caused an increase in the intracellular $Ca^{2+}$ level, which was measured using yellow cameleon-2. These results demonstrate that yellow cameleon-2 can be conveniently used to examine the function of the olfactory receptors expressed in heterologous cells.

• 한국환경농학회지
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• 제40권1호
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• pp.1-12
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• 2021

BACKGROUND: G protein-coupled receptors (GPCRs) are widely distributed in various organisms. Insect GPCRs shown as in vertebrate GPCRs are membrane receptors that coordinate or involve in various physiological processes such as learning/memory, development, locomotion, circadian rhythm, reproduction, etc. This study aimed to identify GPCRs expressed in fat body and compare the expression pattern of GPCRs in different temperature conditions. METHODS AND RESULTS: To identify GPCRs genes and compare their expression in different temperature conditions, total RNAs of fat body in Plutella xylostella larva were extracted and the transcriptomes have been analyzed via next generation sequencing method. From the fat body transcriptomes, genes that belong to GPCR Family A, B, and F were identified such as opsin, gonadotropin-releasing hormone receptor, neuropeptide F (NPF) receptor, muthuselah (Mth), diuretic hormone receptor, frizzled, etc. Under low temperature, expressions of GPCRs such as C-C chemokine receptor (CCR), opsin, prolactin-releasing peptide receptor, substance K receptor, Mth-like receptor, diuretic hormone receptor, frizzled and stan were higher than those at 25℃. They are involved in immunity, feeding, movement, odorant recognition, diuresis, and development. In contrast to the control (25℃), at high temperature GPCRs including CCR, gonadotropin-releasing hormone receptor, moody, NPF receptor, neuropeptide B1 receptor, frizzled and stan revealed higher expression whose biological functions are related to immunity, blood-brain barrier formation, feeding, learning, and reproduction. CONCLUSION: Transcriptome of fat body can provide understanding the pools of GPCRs. Identifications of fat body GPCRs may contribute to develop new targets for the control of insect pests.

• Reproductive and Developmental Biology
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• 제29권1호
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• pp.43-48
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• 2005

돼지 웅성 페르몬인 5a-androst-16-en-3-one을 대체할 수 있는 활성 분자를 탐색하여 가축의 생산과 수요를 조절하기 위한 생물학적 자극 통제 수단으로 활용하고자 냄새 분자로서 2-cyclohexyloxytetrahydrofurane (A) 및 2-phenoxytetrahydrofurane (B) 유도체들의 구조 변화와 수용체인 porcine odorant binding protein (pOBP)에 대한 결합친화력 상수(p[Od.]/sub 50/) 사이의 정량적인 구조-활성관계에 관한 분자 HQSAR 모델XI을 유도하였다. 냄새 분자 중에서 cyclohexyl-치환체(A)가 phenyl-치환체(B)보다 높은 결합 친화력을 나타내었으며(A>B) 모델XI은 분자 조각크기(5∼8), 홀로그램 길이(97 bin)의 키랄성(chirality)조건에서 예측성(q²=0.916)과 상관성(r²=0.988)이 매우 양호하였다. 기여도(contribution map)로부터 냄새 분자의 결합 친화력 상수에 기여하는 부분은 2-oxyfuryl group의 C3 및 C5 원자인 반면에 cyclohexyl 고리상 tert-butyl group의 중심 탄소 원자와 furyl group의 C4 원자 부분은 기여하지 않았다.

• 한국동물생명공학회지
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• 제36권4호
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• pp.292-298
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• 2021

Olfactory receptors (OR) are primarily responsible for the detection of odorant molecules. We previously demonstrated that OR7D4, an OR for androstenone, is expressed in the vomeronasal organ and olfactory epithelium tissue of stallions. Recently, the expression of OR1I1 in the human testes was reported and the possible roles of OR1I1 in the testicular cells were suggested. The objectives of this study were 1) to explore the expression of OR7D4 and OR1I1 in stallion testes, and 2) to define the specific localization of OR7D4 and OR1I1 in the testicular tissues. Stallion testicular tissue samples were used for this study. Western blot was performed to confirm the cross-reactivity of OR7D4 and OR1I1 antibody with stallion testicular tissue samples. OR7D4 and OR1I1 gene expressions were investigated using reverse transcription-polymerase chain reaction (RT-PCR) in stallion testes. Immunofluorescence was performed to investigate the expression of OR7D4 and OR1I1 in stallion testicular tissues. The protein bands for OR7D4 and OR1I1 from the testes were observed at approximately 38 kDa and 43 kDa, respectively. The mRNA of OR7D4 and OR1I1 were detected in stallion testes. Immunolabeling of OR7D4 and OR1I1 in the cytoplasm of both spermatogonia and Leydig cells was observed. In conclusion, androstenone and another odorant chemical, which is recognized by OR1I1, may play an important role in stallion testes.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.169-174
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• 2006

돼지 페르몬성 분자를 탐색하기 위하여 일련의 green odorant로서 기질 분자인 2-(Cyclohexyloxy)tetrahydrofurane 유도체들의 정량적인 구조와 수용체인 porcine odorant binding protein(pOBP) 사이의 결합 친화력 상수($p(Od)_{50}$)에 대한 비교 분자 유사성 지수 분석(CoMSIA)을 실행하였다. 가장 양호한 CoMSIA 모델(I-AI)은 기질 분자내 입체 중심의 절대 배열이 $I:\;C_{1'}(R),\;C_2(S)$인 분자를 atom based fit 정렬하였을 경우의 입체장 조건에서 유도되었으며 PLS 분석 결과, 예측성이 ${r^2}_{cv.}(q^2)=0.856)$ 그리고 적합성이 ${r^2}_{ncv.}=0.964)$ 이었다. 모델의 CoMSIA 등고도 상, pOBP와 냄새 분자 사이의 상호작용으로부터 가장 높은 결합 친화력을 나타내는 분자의 구조적 특정들을 이해할 수 있었다.

• BMB Reports
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• 제45권11호
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• pp.612-622
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• 2012

Olfactory receptors (ORs) detect volatile chemicals that lead to the initial perception of smell in the brain. The olfactory receptor (OR) is the first protein that recognizes odorants in the olfactory signal pathway and it is present in over 1,000 genes in mice. It is also the largest member of the G protein-coupled receptors (GPCRs). Most ORs are extensively expressed in the nasal olfactory epithelium where they perform the appropriate physiological functions that fit their location. However, recent whole-genome sequencing shows that ORs have been found outside of the olfactory system, suggesting that ORs may play an important role in the ectopic expression of non-chemosensory tissues. The ectopic expressions of ORs and their physiological functions have attracted more attention recently since MOR23 and testicular hOR17-4 have been found to be involved in skeletal muscle development, regeneration, and human sperm chemotaxis, respectively. When identifying additional expression profiles and functions of ORs in non-olfactory tissues, there are limitations posed by the small number of antibodies available for similar OR genes. This review presents the results of a research series that identifies ectopic expressions and functions of ORs in non-chemosensory tissues to provide insight into future research directions.

• 한국식품위생안전성학회지
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• 제2권2호
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• pp.75-81
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• 1987

Bifunctional monoterpene 유도체의 향의 유무는 분자내에 존재하는 두 개의 관능기인, proton donor(AH)와 proton acceptor(B)가 olfaction과 구조활성 상호작용(SAR)과 밀접한 관계가 있다. 일반적으로 Ohloff가설에 의하면, p-menthane monoterpene 분자내의 AH와 B의 입체배위적 최소거리가 3${\AA}$ 이하인 경우 향을 갖고, 3${\AA}$ 이상인 경우는 향을 갖지 않는다. Bifunctional pinanone, thujane, carane, carvomenthone 및 기타 menthone 유도체 등을 이용하여 이 가설을 확대 연구하였다. Bifunctional monoterpene인 (원문이미지참조) 등은 분자내에 각각 AH (OH 혹은 COOH)와 B (C=O)의 입체배위적 최소 거리가 항상 3${\AA}$ 이하여서 향을 가지며, 이들은 olfactory three point attachment에 의한 구조활성 상관관계를 가지는 것으로 사료된다. 상기 화합물의 proton donor인 OH, 혹은 COOH가 각각 acetylation이나 methylation되는 경우에는 proton donor로서의 기능 상실로 향이 사라지게 되었다.