• Journal of Veterinary Science
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• 제25권5호
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• pp.63.1-63.12
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• 2024

Importance: Glutamic acid decarboxylase 67 (GAD67) is a gamma-aminobutyric acid (GABA) synthesis enzyme associated with the function of other neurotransmitter receptors, such as the N-methyl-D-aspartate (NMDA) receptor and cannabinoid receptor 1. However, the role of GAD67 in the development of different abused drug-induced reward behaviors remains unknown. In order to elucidate the mechanisms of substance use disorder, it is crucial to study changes in biomarkers within the brain's reward circuit induced by drug use. Objective: The study was designed to examine the effects of the downregulation of GAD67 expression in the dorsal striatum on reward behavior development. Methods: We evaluated the effects of GAD67 knockdown on depression-like behavior and anxiety using the forced swim test and elevated plus maze test in a mouse model. We further determined the effects of GAD67 knockdown on ketamine- and JWH-018-induced conditioned place preference (CPP). Results: Knockdown of GAD67 in the dorsal striatum of mice increased depression-like behavior, but it decreased anxiety. Moreover, the CPP score on the NMDA receptor antagonist ketamine was increased by GAD67 knockdown, whereas the administration of JWH-018, a cannabinoid receptor agonist, did not affect the CPP score in the GAD67 knockdown mice group compared with the control group. Conclusions and Relevance: These results suggest that striatal GAD67 reduces GABAergic neuronal activity and may cause ketamine-induced NMDA receptor inhibition. Consequently, GAD67 downregulation induces vulnerability to the drug reward behavior of ketamine.

• BMB Reports
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• 제38권5호
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• pp.595-601
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• 2005

In this study, we have isolated a rice (Oryza sativa L.) glutamate decarboxylase (RicGAD) clone from a root cDNA library, using a partial Arabidopsis thaliana GAD gene as a probe. The rice root cDNA library was constructed with mRNA, which had been derived from the roots of rice seedlings subjected to phosphorus deprivation. Nucleotide sequence analysis indicated that the RicGAD clone was 1,712 bp long, and harbors a complete open reading frame of 505 amino acids. The 505 amino acid sequence deduced from this RicGAD clone exhibited 67.7% and 61.9% identity with OsGAD1 (AB056060) and OsGAD2 (AB056061) in the database, respectively. The 505 amino acid sequence also exhibited 62.9, 64.1, and 64.2% identity to Arabidopsis GAD (U9937), Nicotiana tabacum GAD (AF020425), and Petunia hybrida GAD (L16797), respectively. The RicGAD was found to possess a highly conserved tryptophan residue, but lacks the lysine cluster at the C-proximal position, as well as other stretches of positively charged residues. The GAD sequence was expressed heterologously using the high copy number plasmid, pVUCH. Our activation analysis revealed that the maximal activation of the RicGAD occurred in the presence of both $Ca^{2+}$ and calmodulin. The GAD-encoded 56~58 kDa protein was identified via Western blot analysis, using an anti-GAD monoclonal antibody. The results of our RT-PCR analyses revealed that RicGAD is expressed predominantly in rice roots obtained from rice seedlings grown under phosphorus deprivation conditions, and in non-germinated brown rice, which is known to have a limited phosphorus bioavailability. These results indicate that RicGAD is a $Ca^{2+}$/calmodulin-dependent enzyme, and that RicGAD is expressed primarily under phosphate deprivation conditions.

• 동의생리병리학회지
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• 제28권6호
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• pp.614-620
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• 2014

Kainic acid (KA) is a excitatory agonist causing epileptic seizure and excitotoxicity in the hippocampus. Gastrodia Elata (GE) is known to have anti-convulsant and anti-oxidant effects. This study was investigated a possible role of GE in suppressing epileptic seizure using KA-induced epilepsy mouse model. Eight-week-old male C57BL/6 mice were administrated GE (50 or 500 mg/kg) once a day for 5 days, and then injected KA (30 mg/kg) intraperitoneally. Behavioral changes in mice by KA were evaluated for 90 minutes immediately after the KA administration. Six hours after the KA administration, their brains were harvested and the expressions of glutamate decarboxylase 67 (GAD-67) and K+-Cl- cotransporter 2 (KCC2) in the hippocampus of the mice were measured by immunohistochemistry.GE delayed the onset of epileptic seizure after KA administration, suppressed the severity of the seizure and decreased the number of severe seizures dose dependently. Moreover, GAD-67 and KCC2 expressions in the cornu ammonis (CA) 1 and CA3 of 500 mg/kg GE administrated mice were significantly increased compared to those in KA-treated mice.GAD-67 and KCC2 play an important role in regulating GABAergic system. Our results suggest that GE has anti-convulsant effect against KA-induced epileptic seizure through enhancing GABAergic system.

• Korean Journal of Acupuncture
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• 제24권4호
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• pp.99-110
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• 2007

Objectives : Epilepsy is one of the most common serious brain disorders that affect people of all ages, and it is characterized by recurrent unprovoked seizures. We examined whether acupuncture can reduce both the incidence of seizures and hippocampal cell death in dentate gyrus (DG) using a mouse model of kainic acid (KA)-induced epilepsy. Methods : ICR mice ($20{\sim}25$ g) were given acupuncture once a day at acupoint HT8 (sobu) bilaterally during 2 days before KA injection. After an intracerebroventricular injection of 0.1${\mu}g$ of KA, acupuncture treatment was subsequently administered once more (total 3 times), and the degree of seizure was observed for 20 min. Three hours after injection, we confirmed the neural cell death using cresyl violet staining and silver impregnation staining, and determined the expressions of c-Fos and glutamate decarboxylase (GAD)-67 using immunohistochemistry techniques in the DG. Results : KA induced epileptic seizure, neural cell death, increased c-Fos expression and decreased GAD-67 expression in the DG. Acupuncture treatment at HT8 reduced the severity of the epileptic seizure and inhibited neural cell death from KA. In addition, acupuncture normalized the expressions of c-Fos and GAD-67 in the same areas. Conclusions : These results demonstrated that acupuncture treatment at HT8 may reduce the KA-induced epileptic seizure and neural cell death in the DG possibly by normalizing c-Fos expressions and the gamma-aminobutyric acid neurons.

• Clinical and Experimental Pediatrics
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• 제61권5호
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• pp.150-155
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• 2018

Purpose: Infantile spasms, also known as West syndrome, is an age-specific epileptic seizure. Most patients with this condition also exhibit delayed development. This study aimed to determine the effect of long-term prenatal stress on susceptibility to infantile spasms. Methods: We subjected pregnant rats to acute or chronic immobilization stress. Resulting offspring received N-methyl-D-aspartic acid (15 mg/kg, intraperitoneally) on postnatal day 15, and their behaviors were observed 75 minutes after injection. The expression of KCC2 and GAD67 was also determined using immunohistochemistry. Results: Exposure to long-term prenatal stress increased the frequency of spasms and decreased the latency to onset of spasms compared with offspring exposed to short-term prenatal stress. Expression of KCC2 and GAD67 also decreased in the group exposed to long-term prenatal stress compared with the group exposed to short-term prenatal stress. Conclusion: Our study suggests that exposure to long-term prenatal stress results in increased susceptibility to seizures.

• 한국양식학회지
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• 제3권1호
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• pp.65-77
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• 1990

피조개의 성장 및 적정 양식장의 개발에 관한 기초자료를 얻기 위하여 거제군 사곡리 앞바다에서 천연채묘한 치패를 사용하여 가덕도 및 남해의 피조개 양성장에서 성장비교 실험한 결과는 다음과 같다. 실험개시시의 각장 1.38$\pm$0.32cm와 전중 3.39$\pm$0.30g의 피조개 치패를 17개월간 양성한 결과, 각각의 성장은 가덕도 6.41$\pm$0.30cm, 67.58$\pm$9.63g, 남해 7.20$\pm$0.30cm, 97.33$\pm$11.45g으로 자라나, 남해가 가덕도에 비하여 피조개의 성장이 좋았다. 두 양성장의 피조개 비만도는 가덕도에서 8$\~$9월, 남해에서는 3$\~$5월에 최대치를 보였다. 실험기간중 수온 및 비중은 가덕도에서 $4.9\~27.1^{\circ}C$, 1.0147$\~$1.0259, 남해에서는 $4.5\~26.3^{\circ}C$, 1.0191$\~$1.0263으로, 특히 비중에 있어서 가덕도가 남해에 비하여 낮았다. 입도분석에 의한 저질의 왜도는 남해에서 positive skewness를 나타낸 반면, 가덕도에서는 negative skewness를 보여 가덕도 앞바다의 조류는 남해보다 강한 것으로 나타났다. 양성장간의 성장비교 및 환경 분석에 의한 남해 미조리 지선어장이 가덕도 장항 지선어장에 비하여 조건이 좋은 양성장인 것으로 판단된다.

• Journal of Korean Biological Nursing Science
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• 제22권4호
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• pp.279-287
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• 2020

Purpose: Some of the adolescent drinks more sugar-sweetened beverages. However, there is little evidence on the effect of eating behavior on emotional state and neurochemical changes under stress, especially on the levels of typical inhibitory neurotransmitters and gamma-aminobutyric acid. This article demonstrates that sucrose or saccharin drink reduces stress-related behavior responses and GABAergic deficits in adolescent rats. Methods: We randomly assigned 7-weeks-old Sprague-Dawley male rats to three groups: control group (Control), restraint stress only group (Stress), and restraint stress with unrestricted access to saccharin solution (Saccharin) and sucrose solution (Sucrose) as a positive control. We evaluated both anxious and depressive moods using an open field test and forced swim test, respectively. Using western blot analyses, the expression of a GABA-synthesizing enzyme, glutamate decarboxylase-67 (GAD67) and GABAergic markers, including calbindin and parvalbumin was assessed in the prefrontal cortex and the amygdala. Results: We found that both the drinks alleviated anxiety and depressive moods, induced significant attenuation in GAD67 level, and reduced calbindin level under stress in the prefrontal cortex and the amygdala. Conclusion: The results provide an understanding of the effect of sucrose or saccharin drink on stress-related responses. We propose the consumption of sweet drinks as a plausible strategy to alleviate stress-related alterations in adolescents.

• The Korean Journal of Physiology and Pharmacology
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• 제27권1호
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• pp.113-125
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• 2023

It has been reported that stressful events in early life influence behavior in adulthood and are associated with different psychiatric disorders, such as major depression, post-traumatic stress disorder, bipolar disorder, and anxiety disorder. Maternal separation (MS) is a representative animal model for reproducing childhood stress. It is used as an animal model for depression, and has well-known effects, such as increasing anxiety behavior and causing abnormalities in the hypothalamic-pituitary-adrenal (HPA) axis. This study investigated the effect of MS on anxiety or aggression-like behavior and the number of GABAergic neurons in the hippocampus. Mice were separated from their dams for four hours per day for 19 d from postnatal day two. Elevated plus maze (EPM) test, resident-intruder (RI) test, and counted glutamic acid decarboxylase 67 (GAD67) or parvalbumin (PV) positive cells in the hippocampus were executed using immunohistochemistry. The maternal segregation group exhibited increased anxiety and aggression in the EPM test and the RI test. GAD67-positive neurons were increased in the hippocampal regions we observed: dentate gyrus (DG), CA3, CA1, subiculum, presubiculum, and parasubiculum. PV-positive neurons were increased in the DG, CA3, presubiculum, and parasubiculum. Consistent with behavioral changes, corticosterone was increased in the MS group, suggesting that the behavioral changes induced by MS were expressed through the effect on the HPA axis. Altogether, MS alters anxiety and aggression levels, possibly through alteration of cytoarchitecture and output of the ventral hippocampus that induces the dysfunction of the HPA axis.

• Natural Product Sciences
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• 제18권2호
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• pp.67-75
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• 2012

Zizyphi Spinosi Semen (ZSS) have been widely used for the treatment of insomnia in Asia. This experiment was performed to investigate whether methanol extract of ZSS (MEZSS) has hypnotic effects through the ${\gamma}$-amino butyric acid (GABA)ergic systems. MEZSS inhibited the locomotor activity. MEZSS enhanced pentobarbital-induced sleep behaviors. However, MEZSS itself did not induce sleep at higher dose, similar to muscimol. On the other hand, both pentobarbital and MEZSS increased the non rapid eye move (NREM) sleep, especially reducing the -wave electroencephalogram (EEG) activity in REM sleep. MEZSS showed similar effects with muscimol on potentiating chloride influx induced by pentobarbital. MEZSS significantly increased GABAA receptors ${\gamma}$-subunit expression and slightly decreased ${\beta}$-subunit expression in hypothalamus and thalamus, showing that subunit-expression was similar to diazepam. In addition, MEZSS enhanced the expression of glutamic acid decarboxylase (GAD). In conclusion, it is suggested that MEZSS might augment pentobarbital-induced sleep behaviors through the modification of GABAergic systems.

• 대한화장품학회지
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• 제50권2호
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• pp.179-192
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• 2024

본 연구에서는 천연물 6 종 앵두, 으름, 살구, 야콘, 금귤 및 핑거라임을 활용하여 스트레스 완화에 의한 수면 개선 및 피부장벽 개선 효과를 검증하였다. 실험 결과, 6 종의 천연물 중 앵두, 핑거라임, 으름 및 금귤이 유의적으로 스트레스 호르몬인 cortisol의 생성을 억제하였다. 또한, 수면 조절에 관여하는 GABA 생성 효소인 GAD67의 발현은 앵두 정제수 추출물 및 핑거라임 50% 에탄올 추출물에서 유의한 증가 효과를 보였으며 앵두 용매별 추출물은 총 폴리페놀의 함량이 가장 높게 확인되었다. 스크리닝 결과 활성이 가장 높은 앵두 추출물을 활용하여 후속 실험을 진행하였다. 앵두 용매별 추출물 중, 30% 에탄올 추출물이 가장 높은 DPPH radical 소거 활성을 보였으며, 정제수 추출물은 GABA 생성 및 피부 장벽 인자인 filaggrin과 claudin-1의 발현을 가장 높게 증가시켰다. HPLC 분석을 통해 앵두 추출물의 주성분으로 quercitrin이 확인되었고 추출 용매별 quercitrin 함량은 30% 에탄올 > 정제수 > 70% 에탄올 > 50% 에탄올의 추출 용매 순으로 높은 함량을 보였다. Quercitrin은 cortisol의 생성을 농도의존적으로 억제하였으며, cortisol에 의해 감소되었던 GAD67 발현 및 GABA 생성을 유의적으로 증가시켰다. 연구를 통해 앵두 추출물이 스트레스를 완화시켜 수면 개선 및 피부 장벽 강화에 도움을 주는 화장품 소재로서 활용이 가능함을 입증하였다.