흰쥐의 삼차신경절에서 Transient receptor potential ankyrin 1 (TRPA1)의 발현 특성에 관한 연구

백상규,나연경,김윤숙,Paik, Sang-Kyoo,Na, Yeon-Kyung,Kim, Yun-Sook 한국현미경학회 2012 Applied microscopy Vol.42 No.1

Transient receptor potential ankyrin 1 (TRPA1)은 $17^{\circ}C$보다 낮은 유해한 온도 및 자극적인 화합물에 의해 활성화되며 통각을 조절한다. 그러나 TRPA1에 의한 통각정보가 어떻게 처리되는지에 대한 정보는 많이 알려져 있지 않다. 본 연구에서는 흰쥐의 삼차신경절에서 TRPA1을 발현하는 신경세포의 특성을 규명하기 위해서, 면역형광기법을 사용하여 TRPA1을 발현하는 신경세포에서 다른 통각수용기들에서 발현되며, 특징적인 기능을 수행하는 수용기인 transient receptor potential vanilloid 1 (TRPV1)와 $P2X_3$와의 발현양상을 조사하였다. TRPA1을 발현하는 신경세포에서 열감각수용기이며, 통각표지자인 TRPV1과의 공존을 조사해 본 결과, TRPA1 면역양성 신경세포 중에서 58.8% (328/558)가 TRPV1을 동시에 발현하였으며, 41.2% (230/558)가 TRPA1만 발현하고 TRPV1을 발현하지 않았다. TRPA1을 발현하는 신경세포 중 TRPV1을 동시에 발현하는 신경세포는 대부분 작거나 중간크기였다. 또한 TRPA1과 조직의 손상, 그리고 염증 시 분비되는 ATP와 결합하는 $P2X_3$와의 공존을 조사해 본 결과, TRPA1 면역양성 신경세포 중에서 26.1% (310/1186)의 신경세포에서 $P2X_3$을 동시에 발현하였으며, 73.9% (876/1186)의 신경세포에서 TRPA1만 발현하였다. TRPA1을 발현하는 신경세포 중 $P2X_3$을 동시에 발현하는 신경세포는 대부분 작거나 중간크기였다. 이러한 결과는 TRPA1을 발현하는 신경세포가 TRPV1 또는 $P2X_3$를 동시에 발현함으로써 동일한 신경세포가 구강안면영역에서의 냉통각 및 열통각을 조절할 뿐 아니라, 냉통각 및 염증성동통을 동시에 전달하는 등 하나의 신경세포가 여러 가지 통각의 전달에 관여하는 것을 시사한다. Transient receptor potential ankyrin 1 (TRPA1), responding to noxious cold (${\leq}17^{\circ}C$) and pungent compounds, is implicated in nociception, but little is known about the coexpression of TRPA1 and other channels or receptors involved in the nociception in craniofacial regions. To address this issue, we characterized the TRPA1-immunopositive (+) neurons in the rat trigeminal ganglion (TG) and investigated their colocalization with other proteins known to be expressed in nociceptive neurons, such as transient receptor potential vanilloid (TRPV1) and $P2X_3$ receptor, using light microscopic immunofluorescence labeling method with TRPA1 and TRPV1 or $P2X_3$ antisera. The majority of TRPA1+ neurons costained for TRPV1 (TRPV1+/TRPA1+; 58.8%, 328/558) and 41.2% only expressed TRPA1 but not TRPV1. The TRPV1+/TRPA1+ neurons were small and medium sized. In addition, we investigated the colocalization of TRPA1 with $P2X_3$, a nonselective cation channel activated by ATP that may be released in the extracellular space as a result of tissue damage and inflammation. Among all TRPA1+ TG neurons, 26.1% (310/1186) costained for $P2X_3$, whereas 73.9% (876/1186) of TRPA1+ neurons did not coexpress $P2X_3$. $P2X_3$+/TRPA1+ neurons were predominantly small and medium sized. These results suggest that TRPA1+ neurons coexpressing TRPV1 or $P2X_3$ are involved in specific roles in the transmission and processing of orofacial nociceptive information by noxious cold, heat, and inflammation.

발효, 세포배양, 생물공정 Saccharomyces cerevisiae KNU5377의 NaCl에 대한 적응이 고온내성과 알코올발효에 미치는 영향

백상규 ( Sang Kyoo Paik ),윤혜선 ( Hae Sun Yun ),사금희 ( Keum Hee Sa ),김일섭 ( II Sup Kim ),이인구 ( In Koo Rhee ),박희동 ( Heui Dong Perk ),유춘발 ( Choon Bal Yu ),진익렬 ( Ingn Yol Jin ) 한국미생물생명공학회 2003 한국미생물·생명공학회지 Vol.31 No.1

삼차신경중간핵에서 저역치기계자극수용기 유래 들신경섬유 종말의 연접양상

안형준,백상규,배용철,최종훈,김종열,Ahn, Hyoung-Joon,Paik, Sang-Kyoo,Bae, Yong-Chul,Choi, Jong-Hoon,Kim, Chong-Youl 대한안면통증구강내과학회 2005 Journal of Oral Medicine and Pain Vol.30 No.1

In order to evaluate the mechanism of transmission as well as processing of sensory information originating from low-threshold mechanoreceptor in oral and maxillofacial region at primary synaptic region of trigeminal nervous system, vibrissa afferent fibers of adult cat were labeled with intra-axonal HRP injection. Serial sections containing labeled boutons were obtained from the piece of trigeminal interpolar nucleus. Under electron microscope, total 30 labeled boutons were observed, and ultrastructural characteristics, frequency of occurence, synaptic organizations of vibrissa afferent terminals were analysed. The results were as follows: 1. Labeled boutons contained clear, spherical synaptic vesicles with diameter of 45$\sim$55nm. They formed asymmetrical synapse with dendrites showing definite postsynaptic density, larger synaptic cleft, multiple synaptic structures at various regions. With unlabeled axon terminals(p-ending) containing polymorphic synaptic vesicles, they formed symmetrical synapse showing indefinite postsynaptic density and narrower synaptic area. 2. Each labeled bouton formed 1 to 15 synapses, the average of 4.77$\pm$3.37 contacts per labeled bouton, with adjacent neuronal profiles. Relatively complex synaptic organization, which formed synapses with more than 5 neuronal profiles, was observed in a large number(46.7%, n=14) of labeled boutons. 3. Axo-somatic synapse was not observed. The number of axo-dendritic synapse was 1.83$\pm$1.37 per labeled bouton. Majority(85.0%) of axo-dendritic synapses were formed with dendritic shafts, nonprimary dendrites(n=47, 1.57$\pm$1.38/1 bouton), however, synapses formed with primary dendrites(n=6, 0.20$\pm$0.41/1 bouton) or dendritic spines(n=2, 0.07$\pm$0.25/1 bouton) were rare. 4. 76.7%(n=23) of labeled boutons formed axo-axonic synapse (2.93$\pm$2.36/1 bouton) with p-endings containing pleomorphic vesicles. Synaptic triad, in which p-endings formed synapses with labeled boutons and dendrites adjacent to the labeled boutons simultaneoulsy, were also observed in 60.0%(n=18) of labeled boutons. From the above results, vibrissa afferent terminals of adult cat showed distinctive synaptic organization in the trigeminal interpolar nucleus, thus, suggests their correlation with the function of the trigeminal interpolaris nucleus, which participates in processing of complex sensory information such as two-point discrimination and motivational-affective action. Further studies on physiologic functions such as quantitative analysis on ultrastructures of afferent terminals and nerve transmitters participating in presynaptic inhibition are required.

Cryptococcus laurentii Y-23의 glucoamylase와 exopolygalacturonase의 동시발효에 미치는 유도기질의 영향

김창화(Chang-Hwa Kim),백상규(Sang-Kyoo Paik),윤혜선(Hye-Sun Yun),진익렬(Ingnyol Jin),유춘발(Choon-Bal Yu) 한국식품과학회 2000 한국식품과학회지 Vol.32 No.4

흰쥐 삼차신경감각핵에서 $P2X_3$와 TRPV1의 공존에 관한 연구

문용석,류창현,조이슬,김홍태,박매자,백상규,문제일,김윤숙,배용철,Moon, Yong-Suk,Ryoo, Chang-Hyun,Cho, Yi-Sul,Kim, Hong-Tae,Park, Mae-Ja,Paik, Sang-Kyoo,Moon, Che-Il,Kim, Yun-Sook,Bae, Yong-Chul 한국현미경학회 2008 Applied microscopy Vol.38 No.3

삼차신경계에서 $P2X_3$와 TRPV1 면역양성 일차들신경섬유는 통각정보의 전달에 중요한 역할을 한다. 본 연구에서는 삼차신경절 및 삼차신경꼬리핵에서 $P2X_3$와 TRPV1 면역양성 신경세포의 형태학적 특성 및 투사양식을 이해하기 위하여, 흰쥐 삼차신경절 및 삼차신경꼬리핵에서 $P2X_3$와 TRPV1에 대한 항체를 사용하여 형광면역염색법 및 형태계측학적인 기법을 시행하여 다음과 같은 결과를 얻었다. $P2X_3$ 면역양성 신경세포중 77.4%의 신경세포에서 (1,401/1,810) TRPV1이 동시에 발현되었으며, TRPV1 면역양성 신경세포중 51.9% (1,401/2,698)의 신경세포에서 $P2X_3$와 공존을 보였다. $P2X_3$와 TRPV1에 동시에 면역양성반응을 보이는 신경세포는 중간크기의 신경세포에서 주로 관찰되었으며, $P2X_3$ 혹은 TRPV1 면역양성 신경세포중 아주 작거나 큰 신경세포에서는 공존하지 않았다. 삼차신경꼬리핵에서 $P2X_3$ 면역양성 신경섬유 및 신경종말들은 제1층과 제2층에 분포하는데 주로 제2층의 안쪽부위에서 밀도가 높게 관찰되었으며, TRPV1 면역양성 신경섬유 및 신경종말들은 제1층과 제2층의 바깥쪽에서 밀도가 높게 관찰되었다. $P2X_3$와 TRPV1이 공존하는 신경섬유 및 신경종말들은 제2층의 안쪽과 바깥쪽의 경계부위에서 관찰되었다. 이러한 연구결과는 $P2X_3$와 TRPV1을 동시에 발현하는 신경세포는 구강안면영역에서 통각정보의 처리에 독특한 역할을 수행할 것이라는 것을 시사한다. Trigeminal primary afferents expressing $P2X_3$ or transient receptor potential vanilloid 1 (TRPV1) are involved in the transmission of nociceptive information. In order to characterize $P2X_3$- and TRPV1-immunopositive neurons in the trigeminal ganglion (TG) and trigeminal caudal nucleus (Vc), we performed immunofluorescence experiments using anti-$P2X_3$ and anti-TRPV1 antisera and a morphometric analysis. 77.4% (1,401/1.801) of all the $P2X_3$-postive neurons coexpressed TRPV1 and 51.9% (1,401/2,698) of all the THFV1-immunopositive neurons also costained for $P2X_3$ in the TG. Immunoreactivity for both $P2X_3$ and TRPV1 were present in medium-sized neurons but not in small- and large-sized neurons. $P2X_3$ and/or TRPV1-immunopositive fibers were observed in the primary afferents and their associated axons in the Vc. These fibers and terminals were distributed in the superficial lamina of Vc: $P2X_3$-immunopositive fibers and terminals were distributed in the lamina I and II, expecially in the inner part of lamina II (lamina IIi), whereas TRPV1-immunopositive ones were densely detected in the lamina I and outer part of lamina II (lamina IIo). Immunopositive fibers and terminals for both $P2X_3$ and TRPV1 were observed on the border between lamina IIi and IIo. These results suggest that terminals coexpressing $P2X_3$ and TRPV1 are involved in specific roles in the transmission and processing of orofacial nociceptive information.

고양이 삼차신경꼬리핵에서 저역치기계자극수용기 유래 들신경종말의 연접이전종말이 함유하는 신경전달물질에 대한 정량적 분석

김윤숙,문철주,조진현,배진영,나연경,복혜정,배용철,백상규,Kim, Yun-Sook,Mun, Cheol-Ju,Cho, Jin-Hyun,Bae, Jin-Young,Na, Yeon-Kyung,Bok, Hye-Jeong,Bae, Yong-Chul,Paik, Sang-Kyoo 한국현미경학회 2012 Applied microscopy Vol.42 No.1

이 연구에서는 삼차신경꼬리핵 제 3~4층에서 저역치기계자극정보를 전달하는 일차들신경섬유의 종말과 연접하는 연접이전종말(presynaptic ending; p-ending)들이 어떤 억제성 신경전달물질을 함유하는 지를 분석하고자 하였다. 이를 위해 전기생리학적으로 동정된 고양이콧수염유래 일차들신경종말을 단일 축삭내 HRP주입법으로 표식하였고, GABA와 glycine에 대한 항혈청으로 포매후금입자면역염색법을 시행한 후, 정량적 분석을 실시하였다. 표식종말과 연접하는 16개 p-ending들 중 8개(50%, 8/16) p-ending들은 GABA만을 함유하였으며, 나머지 8개(50%, 8/16) p-ending들은 GABA와 glycine 모두를 함유하는 집단으로 분류할 수 있었다. 또한, 이 두 집단의 p-ending 사이에는 유의한 평균체적의 차이가 보이지 않았으며, 각 p-ending이 함유하는 GABA와 glycine의 상대적 함량은 서로 달랐다. 이러한 결과들은 삼차신경꼬리핵에서 콧수염유래 일차들신경섬유에 의해 전달되는 저역치기계자극정보는 GABA 및 glycine에 의해 연접이전제어(presynaptic modulation)를 받으며, 그 연접이전제어는 각 일차들신경섬유의 종말마다 다르게 나타날 것 이라는 점을 제시한다. The goal of this study was to identify neurotransmitters in endings (p-endings) presynaptic to low-threshold mechanoreceptive vibrissa afferents in the laminae III/IV of cat trigeminal caudal nucleus (Vc). Rapidly-adapting vibrissa afferents were intra-axonally labeled after electrophysiological identification, and postembedding immunogold staining with antisera against ${\gamma}$-aminobutyric acid (GABA) and glycine was performed, followed by quantitative ultrastructural analysis of p-endings presynaptic to the labeled vibrissa afferent terminals. Sixteen p-endings, which are presynaptic to the HRP-labeled vibrissa afferent terminals, were analyzed in this study: Eight p-endings (50%, 8/16) were immunopositive to GABA but immunonegative to glycine (GABA+ p-ending), and remaining 8 p-endings (50%, 8/16) exhibited immunoreactivity to both GABA and glycine. Bouton volume of the p-endings was not significantly different between the two groups. However, the p-endings differed from each other in relative content of GABA and glycine. These findings suggest that low-threshold mechanoreceptive information conveyed through vibrissa afferent at Vc is presynaptically modulated by GABA and/or glycine, and that degree of presynaptic modulation may differ among each vibrissa afferent terminal.

흰쥐의 두힘살근 운동신경세포의 발달과정에 관한 형태학적 분석

김재현(Jae-Hyun Kim),박미화(Mi-Hwa Park),백상규(Sang-Kyoo Paik),마수경(Su-Kyung Ma),백상흠(Sang-Heum Baek),차두원(Duwon Cha) 대한구강악안면외과학회 2005 대한구강악안면외과학회지 Vol.31 No.2

To analyze the developmental changes in soma diameters of digastric motoneurons, wheat-germ agglutinin conjugated horseradish peroxidase(WGA-HRP) was injected into the digastric muscle and visualized the retrogradely HRP-labeled motoneurons through tungstate/tetramethylbenzidine(TMB) and following diaminobenzidine (DAB) reactions. The results obtained from Sprague-Dawley rats at postnatal days 1 (P1), 10 (P10) and 30 (P30) indicated as follows: firstly, soma diameters of digastric motoneurons showed unimodal distribution in all postnatal days examined; secondly, the period of P1 to P10 (period 1) showed about 2 times faster growth rate than that of P10 to P30 (period 2); thirdly, the smallest soma examined in each postnatal day exhibited slower growth rate with that of the largest one (increase ratio in soma diameters from P1 to P30, smallest vs. largest = 1.62 : 1.93); Finally, relative growth rates a day showed again that period 1 had faster growth rate than that of period 2. Consequently, developmental changes in soma diameters of digastric motoneurons resulted in very different growth rates between both periods. This implies that the growth of the soma is almost completing within P10 and thereafter growing slowly. The period 1 and 2 are corresponding to sucking and sucking/masticatory period, respectively. Therefore present study providing morphological changes in soma diameters of digastric motoneurons suggests that both periods and their different growth rates of the motoneurons in each period may closely be related with each other.