흰쥐에서 반복적 스트레스가 GABA성 신경전달에 미치는 영향

김창수,이종범,성형모,시현석,김진성,박형배,정성덕,하정희 大韓神經精神醫學會 2000 신경정신의학 Vol.39 No.1

연구목적 : 일반적으로 생체는 각종 스트레스에 반응하여 적응(adaptation) 기전이 나타난다. 이러한 적응 기전과 관련하여 포유동물의 중요한 억제성 신경전달계인 GABA성 신경전달계의 스트레스성 자극에 대한 반응양상에 대한 연구가 진행되고 있다. 신경전달의 변화양상은 부하된 스트레스의 종류 및 기간에 따라 다양하게 나타나므로 더욱 상세한 연구를 필요로 한다. 방 법 : 본 연구에서는 하루 2시간의 부동화 스트레스를 반복적으로 2주간 부하한 흰쥐의 뇌조직에서 GABA성 신경전달의 변화 양상을 관찰하고자 하였다. 수용체 결합반응을 통하여 GABA 수용체의 각 component별 배위자들의 각 수용체에 대한 결합 반응의 변화 양상을 정상 대조군와 비교하여 보고, 유의한 변화가 있는 부분의 상세한 검정을 위하여 포화결합반응을 통하여 친화력(Kd) 및 최대결합력(Bmax)에서의 변화양상을 검색하였다. 뿐만 아니라 GABA성 신경전달에 관여하는 중요한 신경전달체인 내인성 GABA의 함량과 대사효소인 GABA transaminase의 활성도의 변화양상도 검색하였다. 결 과 : 연구결과 반복적인 부동화 스트레스는 GABA 수용체 효현제인 [³H]muscimol의 흰쥐 뇌조직 GABA 수용체에 대한 결합도에는 별다른 영향을 미치지 않았다. 대뇌피질조직의 GABA 대사효소인 GABA transaminase 활성도를 감소시켰으나, 피질조직외의 다른 조직의 GABA transaminase 활성도나 내인성 GABA의 농도에는 별다른 영향을 미치지 않았다. 반복적인 부동화 스트레스를 부하한 흰쥐의 대뇌피질, 해마 및 시상하부 조직에서는 [³H]flunitrazepam의 수용체 결합도가 감소하였으며, 포화결합 반응결과 수용체의 수가 감소하였음을 확인하였다. 결 론 : 이상의 결과를 요약하면, 흰쥐에게 부하한 반복적인 부동화 스트레스는 뇌조직의 benzodiazepine 수용체의 수를 감소시킴으로써 GABA성 신경전달을 감소시키며, 이러한 GABA성 신경전달계의 변환은 생체에서 스트레스로 인한 행동장애의 한 기전이 될 수 있을 것으로 생각되었다. Objectives : Changes of GABAergic neurotransmission in response to the application of different types of environmental stress have been the subject of research for over two decades. However, the nature of the changes induced by stress appear to show a dependent phenomena on the type and duration of stressor agent employed. Methods : For this reason, this study was performed to observe the effects of repeated stress on the radioligands binding to GABAA/benzodiazepine receptors of discrete brain regions. The author also examined the activity of GABA transaminase and the concentration of endogenous GABA. Male Sprague-Dawley rats, weighing 150-200g were forced to suffer an immobilization stress for 2 hours during 14 consecutive days. Results : Repeated immobilization stress decreased the binding of [3H]flunitrazepam on the benzodiazepine receptor in the cortex, hippocampus and hypothalamus. Saturation experiments followed by scatchard analyses of the results showed decreased density of benzodiazepine receptor and the affinity remained unchanged. Repeated immobilization stress did not affect the binding of [3H]muscimol on the GABAA receptor, the activity of GABA transaminase, and the concentration of endogenous GABA in the brain regions. Conclusions : From these results, it can be concluded that repeated immobilization stress modulated GABAergic neurotransmission via downregulation of the benzodiazepine receptor in the brain.

Effect of Gamma-Aminobutyric Acid on the Gustatory Nucleus Tractus Solitarius in Rats

Kim, Mi-Won,Park, Ha-Ok,Pahng, Mong-Sook,Park, Sang-Won,Kim, Sun-Hun,Jung, Ji-Yeon,Jeong, Yeon-Jin,Kim, Won-Jae Korean Academy of Oral Biology and the UCLA Dental 2005 International Journal of Oral Biology Vol.30 No.3

Gamma-aminobutyric acid (GABA) is known as an inhibitory neurotransmitter in the neurons of the central nervous system. However, its detailed action mechanisms in the rostral gustatory zone of the nucleus tractus solitarius (rNTS) have not been established. The present study was aimed to investigate the distribution, role and action mechanisms of GABA in rNTS. Membrane potentials were recorded by whole cell recordings in isolated brain slices of the rat medulla. Superfusion of GABA resulted in a concentration-dependent reduction in input resistance in the neurons in rNTS. The change in input resistance ws accompanied by response to a depolarizing pulse were diminished by GABA. Superfusion of the slices with either GABA_(A) agonist, muscimol, GABA_(B) agonist, baclofen or GABA_(C) agonist, TACA, decreased input resistance and reduced the nerve activity in association with membrane hyperpolarization. It is suggested that inhibitory signals play a role in sensory processing by the rNTS, in that GABA actions occur through activation of GABA_(A), GABA_(B) and GABA_(C) receptor. These results suggest that GABA has an inhibitory effect on the rNTS through an activation of GABA_(A), GABA_(B) and GABA_(C) receptors and that the GABAergic inhibition probably plays an important role in sensory processing by the rNTS.

Immunocytochemical Localization of GABA Transporter-3 in the Guinea Pig Retina

이은진(Eun-Jin Lee),오수자(Su-Ja Oh),정진웅(Jin-Woong Chung),천명훈(Myung-Hoon Chun) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.2

망막을 포함하는 중추신경계통에서 억제성 신경전달물질로 작용하고 있는 GABA의 분비와 흡수에 GABA 운송물질이 중요한 역할을 하며, 이 물질은 최소한 3개의 아형으로 구분되고 있다. 이중 GABA 운송물질-1은 주로 신경세포에, 그리고 GABA 운송물질-3은 일부의 신경세포에 분포하되 주로 부챗살아교세포에 존재하는 것으로 흰쥐 및 토끼 망막에서 보고 된 바 있다. 그러나 설치류에 속하면서 비교적 많은 수의 원뿔세포를 함유하고 있는 기니픽 망막에서는 보고된 바 없다. 이 연구에서는 기니픽 망막에서 GABA 운송물질-3를 동정하고 이를 함유한 세포의 신경생화학적 특성을 GABA 운송물 질-3, GABA 합성효소 65, 및 CD15 항체를 이용한 면역세포화학법으로 관찰하였다. GABA 운송물질-3 면역반응은 속핵층에 위치한 일부의 무축삭세포체에서 관찰되었고, 이들 세포의 돌기는 신경절세포 층 가까이의 속얼기층에서 밀집하여 분포하였다. 모든 GABA 운송물질-3 표지세포는 GABA 합성효소65에 면역반응을 나 타내었다. 또한, GABA 운송물질-3 표지 무축삭세포 중 67%는 CD15 면역반응을 나타내었다. 이상의 결과에서 기니픽 망막에서 GABA 운송물질-3는 속얼기층에서 A17 무축삭세포를 통해 암전도로를 조절할 것으 로 생각된다. The cellular localization of the GABA transporter-3 (GAT-3) was examined in the guinea pig retina by immunocytochemistry, using antisera against GAT-3. GAT-3 immunoreactivity was localized to cell bodies in the inner nuclear layer, and labeled processes were densely distributed in the inner plexiform layer (IPL) close to the ganglion cell layer. All GAT-3 labeled cells exhibited GAD65 immunoreactivity. In addition, 67% of GAT-3 labeled amacrine cells showed carbohydrate epitope CD15 immunoreactivity. These results indicate that GAT-3 is involved in modulating the rod pathway in the IPL of the guinea pig retina via presumptive A17 amacrine cells.

Mono sodium glutamate (MSG) 발효 GABA의 수면유도 효과

김승섭(Seung-Seop Kim),오성호(Sung-Ho Oh),정명훈(Myoung-Hoon Jeong),조석철(Seok-Cheol Cho),국무창(Moo-Chang Kook),이석호(Seok-Ho Lee),변유량(Yu-Ryang Pyun),이현용(Hyeon-Yong Lee) 한국식품과학회 2010 한국식품과학회지 Vol.42 No.2

미배아 발효로부터 얻은 GABA 시료를 이용하여 양성대조군으로 사용된 우유와 doxylamine succinate, 음성대조군으로 사용된 caffeine, 일반대조군으로 사용된 증류수와 함께 실험군인 GABA 시료를 농도별로 투여하여 실험을 수행하였다. 멜라토닌 함량을 측정한 결과, 대조군의 2.607±0.410 pg/㎖에 비해 GABA 60 ㎎/㎖ 투여군은 3.243±0.154 pg/㎖, GABA 120 ㎎/㎖ 투여군은 3.425±0.182 pg/㎖, 우유에 녹인 GABA 120 ㎎/㎖ 투여군은 3.464±0.205 pg/㎖으로 나타났다. 반면에 GABA 시료의 섭취 기간에 따른 멜라토닌 호르몬의 누적효과는 유의성이 없었다. 세로토닌 함량을 측정한 결과 대조군의 4.73±0.67 ng/mL에 비해 GABA 60 ㎎/㎖ 투여군은 4.71±1.22 ng/㎖, GABA 120 ㎎/㎖ 투여군은 5.37±0.96 ng/㎖, 우유에 녹인 GABA 120 ㎎/㎖ 투여군은 6.34±0.59 pg/㎖으로 대조군의 평균값과 같거나 비교적 높은경향을 보였으며 농도별로 유의성이 있음을 확인하였다. 특히, GABA 120 ㎎/㎖ 투여시 멜라토닌과 세로토닌 모두 시판되는 수면보조제보다 높은 것으로 나타났다. Relatively large amounts of GABA can be produced by the fermentation of rice bran. Therefore, this study was conducted to investigate the effects of GABA on the secretion of melatonin and serotonin for the development of a sleep inductive compound. The secretion levels of melatonin and serotonin from mice were found to be 3.425±0.182 pg/㎖ and 5.37±0.963 ng/㎖, respectively, in response to feeding 120 ㎎/㎖ of GABA while they were 2.607±0.41 pg/㎖ in the control. The secretion of both melatonin and serotonin was increased up to the 13.51% and 34.99%, respectively, when compared to the negative control. However, the feeding of milk alone did not have a great effect on the melatonin and serotonin secretions. Conversely, feeding of milk with GABA enhanced the secretion of serotonin. The amounts of both melatonin and serotonin secreted increased with respect to the increase in GABA concentrations during feeding. Interestingly, the induction level of melatonin was relatively higher than that of serotonin in response to feeding 120 ㎎/㎖ of GABA. This is the first study to report that GABA has an ability to induce sleep related hormones in mice; therefore, it has the potential for use as a natural sleep aid.

보리 잎과 옥수수 수염의 혼합과 유산균 발효를 이용한 γ-aminobutyric acid 생산 증진

김형주,윤영걸 한국유기농업학회 2017 韓國有機農業學會誌 Vol.25 No.1

GABA는 glutamic acid decarboxylase에 의해서 L-glutamic acid가 탈탄산화되어 생합성된 비단백질 아미노산이다. GABA는 식물에서 스트레스에 대항한 대응반응으로 생성된다. 사 람의 중추신경계에서는 주요 억제성 신경전달물질 중 하나로 항고혈압, 항당뇨 효능이 있 다고 알려져 있다. 본 연구에서 우리는 보리 잎과 옥수수 수염을 유산균과 함께 발효함으 로써 GABA 생성을 증진시키고자 하였다. 보리 잎과 옥수수 수염을 다양한 무게 비율로 조 합하여 혼합하였고, 30℃에서 48시간 동안 배양기 안에서 L. plantarium과 함께 발효시켰다. 발효된 혼합물을 열수 추출한 후, thin layer chromatography와 GABase assay를 이용하여 GABA의 생산을 분석하였다. 우리는 9:1 혼합발효추출물이 다른 비율의 추출물 보다 GABA 함량이 높은 것을 확인하였는데 이것은 혼합과 발효기술이 보리 잎과 옥수수 수염 내 GABA 양 증진에 효과가 있음을 의미한다. 또한 몇 가지 생리활성을 분석한 결과 혼합 발효추출물의 항산화 효능이 비발효 추출물에 비하여 증진되었고 세포독성은 나타나지 않 음을 확인하였다. 이러한 결과는 보리 잎과 옥수수 수염의 조합과 이것을 유산균과 함께 발효시키는 방법이 고함량의 GABA와 증진된 생리 활성을 지닌 기능성 식품으로서의 개발 가능성이 있음을 의미한다. γ-aminobutyric acid (GABA) is a non-proteinogenic amino acid biosynthesized through decarboxylation of L-glutamic acid by glutamic acid decarboxylase. GABA is believed to play a role in defense against stress in plants. In humans, it is known as one of the major inhibitory neurotransmitters in the central nervous system, exerting anti-hypertensive and anti-diabetic effects. In this report, we wanted to enhance the GABA production from the barley leaf and corn silk by culturing them with lactic acid bacteria (LAB). The barley leaf and corn silk were mixed with various weight combinations and were fermented with Lactobacillus plantarum in an incubator at 30℃ for 48 h. After extracting the fermented mixture with hot water, we evaluated the GABA production by thin layer chromatography and GABase assay. We found that the fermented mixture of the barley leaf and corn silk in a nine to one ratio contained a higher level of GABA than other ratios, meaning that the intermixture and fermentation technique was effective in increasing the GABA content. We also tested several biological activities of the fermented extracts and found that the extracts of the fermented mixture showed improved antioxidant activities than the non-fermented extracts and no indication of cytotoxicity. These results suggest that our approach on combining the barley leaf and corn silk and its fermentation with LAB could lead to the possibility of the development of functional foods with high levels of GABA content and improved biological activities.

GABA_(A)α6 유전자 다형성에 따른 알코올 금단 증상의 발현 차이

한덕현,최정은,이병용,김영훈,김혜원,이혜경 大韓神經精神醫學會 2005 신경정신의학 Vol.44 No.2

Introduction : The gamma-aminobutyric acid type A (GABA_(A)) receptor is an important pharmacological target of alcohol.The phamacological characteristics of the receptor are largely determined by its subunit composition, Compared with all othera subtypes, the α 6- containing receptors are more sensitive to GABA and less sensitive to benzodiazepines. The purpose of thisstudy was to address a role for GABA_(A) α 6 receptor subunit gene in the development of alcohol dependence. The differentialmanifestation of alcohol withdrawal symptoms related to GABA_(A) α 6 polymorphism in patients treating with benzodiazepineswas also examined.Methods : Eighty-seven inpatients with alcohol dependence, and sixty healthy controls were evaluated using CIWA-Ar scale.Each patient was genotyped for GABA_(A) α 6 subunit. Association between GABA_(A) α 6 polymorphism and severity of Withd-rawal symptom were determined.Results : No significant difference was found in GABA_(A) α 6 receptor genetic type and allelic distribution between thealcohol dependent and control subject. Tremor was more severe in CC than TT type. TT type had higher degree of anxiety,agitation and headache than CC type. The GABA_(A) α 6 C allele increased the average score of tremor significantly, and T alleleincreased that of agitation.Conclusion : The results suggested that GABA_(A) α 6 genetic polymorphism was not associated with alcohol dependenceand with severity of alcohol withdrawal symptoms. But in benzodiazepine treated patients, GABA_(A) α 6 polymorphism andallelic type show the difference in severity of each withdrawal symptom. These differences of sevehty are partly responsiblefor the unique pharmacological properties associated with the GABA_(A) α 6 subunit.

Naringenin modulates GABA mediated response in a sexdependent manner in substantia gelatinosa neurons of trigeminal subnucleus caudalis in immature mice

박선아,Nguyen Thao Thi Phuong,박수정,한성규 대한약리학회 2024 The Korean Journal of Physiology & Pharmacology Vol.28 No.1

The substantia gelatinosa (SG) within the trigeminal subnucleus caudalis (Vc) is recognized as a pivotal site of integrating and modulating afferent fiberscarrying orofacial nociceptive information. Although naringenin (4',5,7-thrihydroxy -flavanone), a natural bioflavonoid, has been proven to possess various biological effects in the central nervous system (CNS), the activity of naringenin at the orofacial nociceptive site has not been reported yet. In this study, we explored the influence of naringenin on GABA response in SG neurons of Vc using whole-cell patch-clamp technique. The application of GABA in a bath induced two forms of GABA responses: slow and fast. Naringenin enhanced both amplitude and area under curve (AUC) of GABA-mediated responses in 57% (12/21) of tested neurons while decreasing both parameters in 33% (7/21) of neurons. The enhancing or suppressing effect of narin -genin on GABA response have been observed, with enhancement occurring when the GABA response was slow, and suppression when it was fast. Furthermore, both the enhancement of slower GABA responses and the suppression of faster GABA responses by naringenin were concentration dependent. Interestingly, the nature of GABA response was also found to be sex-dependent. A majority of SG neurons from juvenile female mice exhibited slower GABA responses, whereas those from juvenile males predominantly displayed faster GABA responses. Taken together, this study indicates that naringenin plays a partial role in modulating orofacial nociception and may hold promise as a therapeutic target for treating orofacial pain, with effects that vary according to sex. he substantia gelatinosa (SG) within the trigeminal subnucleus caudalis (Vc) is recognized as a pivotal site of integrating and modulating afferent fiberscarrying orofacial nociceptive information. Although naringenin (4',5,7-thrihydroxy -flavanone), a natural bioflavonoid, has been proven to possess various biological effects in the central nervous system (CNS), the activity of naringenin at the orofacial nociceptive site has not been reported yet. In this study, we explored the influence of naringenin on GABA response in SG neurons of Vc using whole-cell patch-clamp technique. The application of GABA in a bath induced two forms of GABA responses: slow and fast. Naringenin enhanced both amplitude and area under curve (AUC) of GABA-mediated responses in 57% (12/21) of tested neurons while decreasing both parameters in 33% (7/21) of neurons. The enhancing or suppressing effect of narin -genin on GABA response have been observed, with enhancement occurring when the GABA response was slow, and suppression when it was fast. Furthermore, both the enhancement of slower GABA responses and the suppression of faster GABA responses by naringenin were concentration dependent. Interestingly, the nature of GABA response was also found to be sex-dependent. A majority of SG neurons from juvenile female mice exhibited slower GABA responses, whereas those from juvenile males predominantly displayed faster GABA responses. Taken together, this study indicates that naringenin plays a partial role in modulating orofacial nociception and may hold promise as a therapeutic target for treating orofacial pain, with effects that vary according to sex.

보리 잎과 옥수수 수염의 혼합과 유산균 발효를 이용한 γ-aminobutyric acid 생산 증진

김형주,윤영걸,Kim, Hyung-Joo,Yoon, Young-Geol 한국유기농업학회 2017 韓國有機農業學會誌 Vol.25 No.1

GABA는 glutamic acid decarboxylase에 의해서 L-glutamic acid가 탈탄산화되어 생합성된 비단백질 아미노산이다. GABA는 식물에서 스트레스에 대항한 대응반응으로 생성된다. 사람의 중추신경계에서는 주요 억제성 신경전달물질 중 하나로 항고혈압, 항당뇨 효능이 있다고 알려져 있다. 본 연구에서 우리는 보리 잎과 옥수수 수염을 유산균과 함께 발효함으로써 GABA 생성을 증진시키고자 하였다. 보리 잎과 옥수수 수염을 다양한 무게 비율로 조합하여 혼합하였고, $30^{\circ}C$에서 48시간 동안 배양기 안에서 L. plantarium과 함께 발효시켰다. 발효된 혼합물을 열수 추출한 후, thin layer chromatography와 GABase assay를 이용하여 GABA의 생산을 분석하였다. 우리는 9:1 혼합발효추출물이 다른 비율의 추출물 보다 GABA 함량이 높은 것을 확인하였는데 이것은 혼합과 발효기술이 보리 잎과 옥수수 수염 내 GABA 양 증진에 효과가 있음을 의미한다. 또한 몇 가지 생리활성을 분석한 결과 혼합발효추출물의 항산화 효능이 비발효 추출물에 비하여 증진되었고 세포독성은 나타나지 않음을 확인하였다. 이러한 결과는 보리 잎과 옥수수 수염의 조합과 이것을 유산균과 함께 발효시키는 방법이 고함량의 GABA와 증진된 생리 활성을 지닌 기능성 식품으로서의 개발 가능성이 있음을 의미한다. ${\gamma}$-aminobutyric acid (GABA) is a non-proteinogenic amino acid biosynthesized through decarboxylation of L-glutamic acid by glutamic acid decarboxylase. GABA is believed to play a role in defense against stress in plants. In humans, it is known as one of the major inhibitory neurotransmitters in the central nervous system, exerting anti-hypertensive and anti-diabetic effects. In this report, we wanted to enhance the GABA production from the barley leaf and corn silk by culturing them with lactic acid bacteria (LAB). The barley leaf and corn silk were mixed with various weight combinations and were fermented with Lactobacillus plantarum in an incubator at $30^{\circ}C$ for 48 h. After extracting the fermented mixture with hot water, we evaluated the GABA production by thin layer chromatography and GABase assay. We found that the fermented mixture of the barley leaf and corn silk in a nine to one ratio contained a higher level of GABA than other ratios, meaning that the intermixture and fermentation technique was effective in increasing the GABA content. We also tested several biological activities of the fermented extracts and found that the extracts of the fermented mixture showed improved antioxidant activities than the non-fermented extracts and no indication of cytotoxicity. These results suggest that our approach on combining the barley leaf and corn silk and its fermentation with LAB could lead to the possibility of the development of functional foods with high levels of GABA content and improved biological activities.

흰쥐의 적출 정관 수축성에 대한 γ-Aminobutyric Acid의 억제작용

안기영,권오철,하정희,이광윤,김원준 영남대학교 의과대학 1992 Yeungnam University Journal of Medicine Vol.9 No.2

GABA는 중추신경계의 대표적인 신경전달 물질로서 GABA 수용체 또는 GABA수용체에 작용하여 진정 작용, 항 불안 작용, 및 근이완 작용을 하는 것으로 알려져 있다. 근래에는 말초조직에도 GABA가 존재하며 신경조정인자 혹은 신경전달인자로서 작용한다는 보고가 있다. 정관에 대한 자율신경 지배는 아드레날린성, 콜린성과 비콜린성 비아드레날린성 신경섬유들이 분포하고 있으나, 종 혹은 부위에 따라 이들 구성요소들의 수축성에 대한 기여도가 다른 것으로 알려져 있다. 본 실험에서는 흰쥐의 전립선 부위 정관의 기본장력 및 전기장 유발 수축에 대한 교감 신경성, 부교감 신경성 및 퓨린성 효현제의 영향을 관찰하였으며, 정관의 흥분성 약물 및 전기장 유발 수축에 미치는 GABA및 GABA관련 약물의 영향을 관찰함으로서 정관의 수축운동에 대한 GABA의 작용기전을 규명해 보고자 흰쥐(Sprague-Dawley)의 전립선 부위 정관 절편을 적출 근편 실험조에 현수하고, 등척성 장럭을 측정하여 다음과 같은 결과를 얻었다. 1. GABA, muscimol및 baclofen은 전기장자극(0.2 Hz, 1 msec, 80 V)유발 수축을 농도 의존적으로 억제하였으며 그 효력은 GABA, baclofen 그리고 muscimol순이었다. 2. GABA의 억제작용은 GABA 수용체 길항제인 DAVA에 의하여 길항되었으나, GABA 수용체 길항제인 bicuculline에 의해서는 길항되지 않았다. 3. Baclofen의 억제작용은 DAVA에 의하여 길항되었으나, muscimol의 억제작용은 bicuculline에 의해서는 길항되지 않았다. 4. Norepinephrine과 ATP는 농도 의존적으로 정관의 수축력을 증가 시켰으나 acetylcholine은 영향을 미치지 못하였다. 5. GABA, baclofen 및 muscimol은 정관의 기본장력에는 영향을 미치지 않았으며, GABA는 norepinephrine과 ATP유발 수축에는 영향을 주지 못하였다. 6. 적출 정관을 20초간 전기장 자극을 가하였을 때 ATP수용체 탈감작제인 mATP를 전처치한 경우에는 FPC가 감소되었고, 신경원에서 catecholamine을 고갈시키는 reserpine을 전처치한 경우에는 STC가 감소되었다. 7. GABA는 mATP를 전처치한 군에서는 정관의 전기장 유발 수축에 영향을 주지 못하였 으나, reserpine을 전처치한 군에서는 전기장 유발 수축을 억제하였다. 이상의 결과로 보아 흰쥐정관의 전립선 부위에는 아드레날린성 신경전달기전과 퓨린성 신경전달기전이 동시에 작용하고 있으며, GABA는 주로 신경원의 GABA 수용체를 통하여 ATP의 유리를 억제하는 것으로 사료된다. GABA is an inhibitory neurotransmitter in central nervous system and produce sedative, antianxiety and muscle reaxing effects via GABA receptor or GABA receptor. Recently it is known that GABA is widely distributed throughout peripheral organs and may play a physiological role in certain organ. The vas deferens is innervated by species-difference. These study, therefore, was performed to investegate the mode and the mechanism of action of GABA on the norepiniphrine-, ATP- and electric stimulation-induced contraction of vas deferens of rat. Sprague-Dawley rats were sacrificed by cervical dislocation. The smooth muscle strips were isolated from the prostastic portion and were mounted in the isolated muscle bath. PSS in the bath was aerated with 95/5 % -O₂/CO₂ at 33℃. Muscle tensions were measured by isometric tension transducer and were recorded by biological recording system. 1. GABA, muscimol, a GABA agonist, and baclofen, a GABA agonist inhibited the electric field stimulation(EFS, 0.2Hz, ImSec, 80 V, monophasic square wave)-induced contraction with a rank order-of potency of GABA greater than baclofen greater than muscimol. 2. The inhibitory effect of GABA was antagonized by delta aminovaleric acid(DAVA), a GABA antagonist, but not by bicuculline, a GABA antagonist. 3. The inhibitory effect of baclofen was antagonized by DAVA, but the effect of muscimol was not antagonized by bicuculline. 4. Exogenous norepinephrine(NE) and ATP contracted muscle strip concentration dependently, but the effect of acetylcholine was negligible: and GABA did not affect the NE-and ATP-induced contractions. 5. GABA, baclofen and muscimol did not affect basal tone, and GABA did not affect the NE-and ATP-induced contractionsm 6. EFS-induced contraction was including 2 distinctable components. The first phasic component was inhibited by beta gamma-methylene ATP(mATP), a desensitizing agent of APT receptor and the Second tonic component was reduced by pretreatment of reserpine(3 ㎎/㎏, IP). 7. GABA inhibited the EFS-induced contraction of reserpinized strips, but not the mATP-treated strips. These results suggest that in the prostatic portion of the rat vas deferens, adrenergic and purinergic neurotransmissions are exist, and GABA inhibits the release of ATP via presynaptic GABA receptor on the excitatory neurons.

Production of γ-amino Butyric Acid by Lactic Acid Bacteria in Skim Milk

Jin Young Cha(차진영),Young Rok Kim(김영록),Bo Ram Beck(백보람),Ji Hun Park(박지헌),Cher Won Hwang(황철원),Hyung Ki Do(도형기) 한국생명과학회 2018 생명과학회지 Vol.28 No.2

동해안 지역 수산발효식품과 수산물로부터 다양한 종류의 유산균들을 분리하여 감마아미노낙산(GABA) 활성을 위해 분석을 하였다. 박층크로마토그래피(TLC)를 이용하여 GABA를 생성하는 4개의 균주를 확보하였으며, 16S rRNA sequencing 분석 결과를 통해 FSFL0004, FSFL0005, FSFL0036 균주는 Lactobacillus (Lb.) brevis, 그리고 FGL0007은 Lactococcus (Lc.) lactis에 가장 유사한 것으로 확인하였다. Lb. brevis FSFL0004와 FSFL0005는 발효된 아귀로부터 분리되었고, Lb. brevis FSFL0036는 갈치 젓갈로부터 분리되었으며, Lc. lactis FGL0007 균주는 참가자미의 내장으로부터 분리되었다. 고속액체크로마토그래피(HPLC)를 사용한 정량분석결과를 보면, FSFL0004, FSFL0005, FSFL0036과 FGL0007에서 각각 10,754.37 μg/ml, 13,082.79 μg/ml, 12,290.19 μg/ml, 45.07 μg/ml의 GABA가 생성되었다. GABA가 풍부한 낙농제품의 발효 종균으로서 상용화 실험을 위해 1% MSG를 포함한 탈지방우유에 4개의 균주를 각각 접종하였다. TLC 결과를 보면 4개의 균주 모두가 GABA 생성을 보였다. HPLC 분석 결과를 보면, 4균주 중 Lc. lactis FGL0007이 가장 높은 GABA 생성(431.42 μg/ml)을 보였다. 본 연구의 내용은 GABA가 함유된 유제품 개발의 기반이 될 수 있을 것으로 생각한다. Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for γ-amino butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to 10,754.37 μg/ml, 13,082.79 μg/ml, 12,290.19 μg/ml, and 45.07 μg/ml respectively in 1% monosodium glutamate-supplemented methionyltRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of γ-amino butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production (431.42 μg/ml) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.