Effects of Maternal Ethanol Consumption and Buspirone Treatment on 5-HT_(1A) and 5-HT_(2A) Receptors in Offspring

Kim, Jung-Ae,Gilespie, Roberta A.,Druse, Mary J. 영남대학교 약품개발연구소 1998 영남대학교 약품개발연구소 연구업적집 Vol.8 No.-

In utero ethanol exposure results in a decreased concentration of serotonin (5-HT) in brain regions containing the cell bodies of 5-HT neurons and their cortical projections. The concentration of 5-HT reuptake sites is also reduced in several brain areas. The present study extended prior work by evaluating the effects of chronic maternal ethanol consumption and maternal buspirone treatment on 5-HT_(1A) and 5-HT_(2A) receptors in mutiple brain areas of offspring. Receptors were quantitated early in postnatal development and at an age when the 5-HT networks are normally well-established. Because fetal 5-HT functions as an essential neurotrophic factor, these studies also determined whether treatment of pregnant rats with buspirone, a 5-HT_(1A) agonist, could overcome the effects of the fetal 5-HT deficit and prevent ethanol-associated receptor abnormalities. The results demonstrated that in utero ethanol exposure significantly alters the binding of 0.1 nM [³H]-8-hydroxy-dipropylaminotetralin to 5-HT_(1A) receptors in developing animals. Ethanol impaired the development of 5-HT_(1A) receptors in the frontal cortex, parietal cortex, and lateral septum; these receptors did not undergo the normal developmental increase between postnatal days 19 and 35. The dentate gyrus was also sensitive to the effects of in utero ethanol exposure. 5-HT_(1A) receptors were increased in this region at 19 days. Maternal buspirone treatment prevented the ethanol-associated abnormalities in 5-HT_(1A) receptors in the dentate gyrus, frontal cortex, and lateral septum. Neither maternal ethanol consumption nor buspirone treatment altered the binding of 2 nM [³H]ketanserin to 5-HT_(2A) receptors in the ventral dentate gyrus,dorsal raphe, parietal and frontal cortexes, striatum, substantis nigra, or nucleus accumbens.

생애초기 저산소증으로 인한 신경행동기형

이성수,이서울,강동원,김진수,김동구 대한신경과학회 1998 대한신경과학회지 Vol.16 No.2

정신분열병 환자에서 도파민 D₃ 및 세로토닌 5-HT2a 수용체 유전자의 유전적 다형성 분석

김윤원(Yoon Won Kim),한진영(Jin-Yeong Han),김성환(Seong Hwan Kim),한홍무(Hong Moo Hahn),최병무(Byeong Moo Choe) 대한생물치료정신의학회 2002 생물치료정신의학 Vol.8 No.2

연구목적 : 도파민 D₃ 수용체 유전자(DRD₃)와 세로토닌 5-HT2a 수용체 유전자(5-HT2aR)의 유전적 다형성 분석을 하여 정신분열병과 이들 유전자와의 상관성 여부를 확인하고자 유전학적 연구를 시행하였다. 방법 : 각각 100명의 정신분열병 환자 및 정상 대조군을 대상으로 말초혈로부터 genomic DNA를 분리하여 DRD₃와 5-HT2aR 유전자에 각각 특이 primer를 사용하여 중합효소연쇄반응(polymerase chain reaction : PCR)을 실시하였다. 증폭한 유전자를 각기 제한효소 절편분석반응을 시행하여 유전자의 유전적 다형성 분석을 시행하였으며 이렇게 분류된 유전자형을 환자군과 정상 대조군에서 상호 비교하기 위하여 통계 분석하였다. 결과 : 증폭한 DRD₃ 유전자를 MscI 제한효소로 절단하여 절편분석을 하였을 때 304bp의 band를 보인 군(-/-), 304bp, 206bp 및 98bp의 세 개의 band를 보인 군(+/-), 그리고 206bp와 98bp의 두 개 band를 보인 군(+/+)이 각각 정신분열병 환자군에서 47명,45명, 8명이었고 정상 대조군에서는 61명, 34명, 5명으로서 두 군 간에 통계적으로 유의한 차이는 없었다(p=0.l32). 5-HT2aR 유전자의 MspI 제한효소 다형성 분석에서도 372bp(-/-), 372bp, 216bp와 156bp(+/-), 그리고 216bp와 156bp(+/+)를 보이는 경우가 정신분열병 환자군과 정상 대조군에서 각각 33명, 48명, 19명과 35명, 48명 17명으로 두 군 사이에 통계적 유의성을 보이지 않았다(p=0.918). 결론 : 본 연구결과는 한국인에 있어서 DRD₃ 유전자와 5-HT2aR 유전자는 정신분열병의 원인이 아닐 가능성을 제시한다. Background : Previous family, twin and adoption studies suggested that genetic factors playa major role in the etiology of schizophrenia. However, the mode of inheritance, like that of other common disorders, is complex and it is generally believed that more than one gene is involved in the genetics of schizophrenia. In the present study, the authors examined 100 unrelated Korean schizophrenics and 100 Korean normal controls, all belonging to a single ethnic population, to determine any association between genetic polymorphisms in the dopamine D₃ receptor(DRD₃) gene and the 5-hydroxytryptamine type 2a receptor(5-HT2aR) gene and schizophrenia. Methods : High molecular weight genomic DNA was extracted from whole blood of 100 schizophrenics and 100age- and sex-matched healthy controls. The Mscl polymorphism of DRD₃ gene and the Mspl of 5-HT2aR gene were typed by restriction enzyme digestion after amplification of genomic DNA by polymerase chain reaction. Genotypic distributions were analyzed using the χ² test. Results : With regard to Mscl/DRD₃ gene polymorphism, the presence of 304bp band denoted a (-/-) homozygote, 3 bands at 304bp, 206bp, and 98bp, a (+/-) heterozygote, and 2 bands at 206bp and 98bp, a (+/+) homozygote. In Mspl/5-HT2aR gene polymorphism, a (-/-) homozygote with a 372bp, a (+/-) heterozygote with 3 bands of 372bp, 216bp, and 156bp, a (+/+) homozygote with 2 bands of 216bp and 156bp were identified. No significant positive association between either the DRD₃ genotype or the 5-HT2aR genotype and schizophrenia was observed. Conclusion : Significant association was not observed between schizophrenia and any of the two polymorphisms at the DRD₃ gene and the 5-HT2aR gene. The results suggest that the DRD₃ and 5-HT2aR polymorphisms may not be involved in the genetic susceptibility to the Korean schizophrenics.

Exercise exerts an anxiolytic effect against repeated restraint stress through 5-HT_2A-mediated suppression of the adenosine A_2A receptor in the basolateral amygdala

Leem, Yea-Hyun,Jang, Jee-Hun,Park, Jin-Sun,Kim, Hee-Sun Elsevier 2019 Psychoneuroendocrinology Vol.108 No.-

<P><B>Abstract</B></P> <P>Repeated or chronic stressful stimuli induce emotion- and mood-related abnormalities, such as anxiety and depression. Conversely, regular exercise exerts protective effects. Here, we found that exercise recovered anxiety-like behaviors, as measured using the open field and elevated plus maze tests in an anxiety mouse model. In addition to behavioral improvement, exercise enhanced the synaptic density of the 5-hydroxytryptamine 2A receptor (5-HT<SUB>2A</SUB>R), but not the 5-HT<SUB>1A</SUB>R in the basolateral amygdala (BLA) region in this mouse model. Furthermore, global treatment with a selective 5-HT<SUB>2A</SUB>R antagonist (MDL11930) generated an anxiety phenotype. Thus, synaptic recruitment of 5-HT<SUB>2A</SUB>R in BLA neurons may mediate the anxiolytic effects of exercise. The exercise regimen also reduced adenosine A<SUB>2A</SUB> receptor (A<SUB>2A</SUB>R)-mediated protein kinase A (PKA) activation, and the anxiolytic effect of the exercise was blunted by local activation of A<SUB>2A</SUB>R within the BLA using CGS21680, a selective A<SUB>2A</SUB>R agonist. Particularly, A<SUB>2A</SUB>R-mediated PKA activity was shown to be dependent on 5-HT<SUB>2A</SUB>R signaling in the BLA. These results imply that repeated stress upregulates A<SUB>2A</SUB>R-mediated adenosine signaling to facilitate PKA activation, whereas regular exercise inhibits A<SUB>2A</SUB>R function by increasing 5-HT<SUB>2A</SUB>R in the BLA. Accordingly, this integrated modulation of 5-HT and adenosine signaling, via 5-HT<SUB>2A</SUB>R and A<SUB>2A</SUB>R respectively, may be a mechanism underlying the anxiolytic effect of regular exercise.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Exercise alleviated repeated-stress-induced anxiety-like behaviors. </LI> <LI> Exercise enhanced 5HT<SUB>2A</SUB>R levels and reduced A<SUB>2A</SUB>R levels in the BLA. </LI> <LI> Synaptic A<SUB>2A</SUB>R and PKA activity were modulated by 5HT<SUB>2A</SUB>Rs in the BLA. </LI> <LI> Synaptic 5HT<SUB>2A</SUB>R and A<SUB>2A</SUB>R are crucial for the anxiolytic effect of exercise. </LI> </UL> </P>

Serotonin contracts the rat mesenteric artery by inhibiting 4-aminopyridine-sensitive Kv channels via the 5-HT2A receptor and Src tyrosine kinase

성동준,노현주,김재곤,박상웅,김보경,조하나,배영민 생화학분자생물학회 2013 Experimental and molecular medicine Vol.45 No.12

Serotonin (5-hydroxytryptamine (5-HT)) is a neurotransmitter that regulates a variety of functions in the nervous,gastrointestinal and cardiovascular systems. Despite such importance, 5-HT signaling pathways are not entirely clear. We demonstrated previously that 4-aminopyridine (4-AP)-sensitive voltage-gated Kþ (Kv) channels determine the resting membrane potential of arterial smooth muscle cells and that the Kv channels are inhibited by 5-HT, which depolarizes the membranes. Therefore, we hypothesized that 5-HT contracts arteries by inhibiting Kv channels. Here we studied 5-HT signaling and the detailed role of Kv currents in rat mesenteric arteries using patch-clamp and isometric tension measurements. Our data showed that inhibiting 4-AP-sensitive Kv channels contracted arterial rings, whereas inhibiting Ca2þ-activated Kþ, inward rectifier Kþ and ATP-sensitive Kþ channels had little effect on arterial contraction, indicating a central role of Kv channels in the regulation of resting arterial tone. 5-HT-induced arterial contraction decreased significantly in the presence of high KCl or the voltage-gated Ca2þ channel (VGCC) inhibitor nifedipine, indicating that membrane depolarization and the consequent activation of VGCCs mediate the 5-HT-induced vasoconstriction. The effects of 5-HT on Kv currents and arterial contraction were markedly prevented by the 5-HT2A receptor antagonists ketanserin and spiperone. Consistently, a-methyl 5-HT, a 5-HT2receptor agonist, mimicked the 5-HT action on Kv channels. Pretreatment with a Src tyrosine kinase inhibitor, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, prevented both the 5-HT-mediated vasoconstriction and Kv current inhibition. Our data suggest that 4-AP-sensitive Kv channels are the primary regulator of the resting tone in rat mesenteric arteries. 5-HT constricts the arteries by inhibiting Kv channels via the 5-HT2A receptor and Src tyrosine kinase pathway.

5HT 2A/T102C 다형성과 정신분열병 및 임상아형과의 관련연구

홍현상,백인호,김정진,이창욱,이 철,도규영 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.4

목 적 : 가계연구, 입양연구, 쌍생아연구들은 정신분열병의 원인으로 유전학적 요소가 매우 중요하다는 것을 지적해 주고 있다. 그러나 정신분열병의 유전적인 복잡성으로 인하여 많은 연구들이 시행되었음에도 유전방식이나 원인 유전자를 밝히지는 못하였다. 최근에는 5HT₂a수용체가 비정형항정신병약물의 주된 작용부위하는 것이 밝혀지면서 5HT₂a 수용체의 유전자가 정신분열병의 원인과 관련이 있을 것이라는 관심이 늘어나게 되었다. 따라서 저자들은 한국인 정신분열병환자들을 대상으로 5HT₂a 수용체 T102C 다형성과의 association study를 시행하였다. 방 법 : 환자군은 강남성모병원에 입원했던 DSM-Ⅲ-R의 진단기준에 의한 정신분열병환자로 250명이었으며 평균연령은 30.1±9.3세 이었다. 정신분열병환자를 위한 양성증상과 음성증상척도의 종합척도를 기준으로 점수가 1이상인 경우를 양성군, -1이하인 경우를 음성군으로 하였다. 대조군은 정신과나 신경과 질환의 과거력이나 가족력이 없는 236명의 지원자로 구성하였으며 평균연령은 23.6±3.7세이었다. 다형성과 정신분열병 및 임상양상과 임상아형 사이의 유전적인 관련성의 분석에는 로지스틱 회귀분석과 일원 변량분석을 시행하였으며 통계 프로그램으로는 SPSS 7.5를 사용하였다. 말초혈액에서 DNA를 분리한후 중합효소 연쇄반응으로 증폭하였으며, 증폭산물을 제한효소인 MspI으로 처리하여 유전자형을 판별하였다. 결 과 : 5HT₂a 수용체 T102C 다형성의 대립유전자 빈도와 유전자형 분포는 환자군과 대조군 사이에 유의한 차이를 보여주지 못하였으며, 임상아형과도 유의한 관련이 없는 것으로 나타났다. 그러나 정신분열병환자를 위한 양성증상과 음성증상척도중 음성증상척도는 유전자형에 따른 유의한 차이를 나타내었다(F=3.828, df=2, p=0.023). 결 론 : 본 연구의 결과는 한국인에서 5HT₂a 수용체의 T102C 다형성이 정신분열병의 발생과 임상아형의 결정에는 관련이 없으나 음성증상의 발현에는 관련이 있음을 시사해 준다고 하겠다. Objectives : Family, twin and adoption studies indicate that genetic factors play a crucial role in the etiology of schizophrenia. However, mode of inheritance of schizophrenia is uncertain, and genes for schizophrenia have not yet been identified despite extensive studies due to the complexity of the genetics of schizophrenia. Currently, 5HT₂a receptor gene has attracted considerable interest as a susceptibility gene of schizophrenia since the 5HT₂a receptor has been known as one of the major target sites of atypical neuroleptics. We conducted an association study of T102C polymorphism in the 5HT₂a receptor gene in Korean schizophrenic patients using PCR-RELP method. Methods : Two hundred and fifty biologically unrelated schizophrenic patients meeting DSM-Ⅲ-R criteria from Kangnam St. Mary's Hospital affiliated with Catholic University of Korea were recruited for our study. The patient group consisted of 123 male and 127 female subjects, aged 30.1±9.3years. The controls were volunteers for DNA library of Kangnam St. Mary's Hospital without family history of psychiatric or neurologic illness. The control group consisted of 124 males and 112 females, aged 23.6±3.7years. Amplified genomic DNA was digested by MspI. The significance of genetic association of the polymorphism was estimated by the logistic regression analysis and ANOVA using SPSS 7.5. Results : The allele frequencies and the genotypic distribution of 5HT₂a receptor gene were not significantly different between the patient and control group. In addition the allele frequencies and the genotypes of 5HT₂a receptor gene were not significantly associated with subtype of schizophrenia. However, negative symptom score according to genotype show significant difference(F=3.828 df=2 p=0.023). Conclusion : It is suggested that even if the development and subtype of schizophrenia may not be associated with T102C polymorphism of 5HT₂a receptor in Korean population, T102C polymorphism may be associated with the severity of negative symptom.

한국인 및 중국 한족 정신분열병 환자의 5-HT2A 수용체 유전자 -1438A/G 다형성

이장호,이광철,이승부,오용인,최영근,조아랑,정주호,장환일 大韓神經精神醫學會 2005 신경정신의학 Vol.44 No.1

Objectives : The purpose of the present study was to investigate the association between -1438A/G polymorphism of 5-HT2A receptor gene and schizophrenia in Korean and Han Chinese population. Methods : A sample of 184 Korean patients with schizophrenia and 96 Korean healthy normal controls and 96 Han Chinese patients with schizophrenia and 96 Han-Chinese healthy normal controls were genotyped for a single nucleotide polymorphism with in 5-HT2A receptor gene (promoter region, A-1438G) by Msp I Resthction Fragment Length Polymorphism (RFLP). Results : There was no difference in allelic frequencies and genotype frequencies of -1438A/G polymorphism between Korean schizophrenics and controls (p=0.13) and Han Chinese schizophrenics and controls (p=0.40). Also, -1438A/G Poly-morphism did not show ethnical difference between Korean and Han Chinese controls. The Scale for the Assessment of Negative Symptoms (SANS) scores showed no significant differences between genotypes of -1438A/G polymorphism in both of Korean and Han Chinese schizophrenics. Conclusion : These results suggest that -1438A/G polymorphism of the 5-HT2A receptor gene is not causally related to the development of schizophrenia in Korean and Han Chinese population, and there no ethnic difference between Korean and Han Chinese population.

Structure-Activity Relationship and Evaluation of Phenethylamine and Tryptamine Derivatives for Affinity towards 5-Hydroxytryptamine Type 2A Receptor

( Shujie Wang ),( Anlin Zhu ),( Suresh Paudel ),( Choon-gon Jang ),( Yong Sup Lee ),( Kyeong-man Kim ) 한국응용약물학회 2023 Biomolecules & Therapeutics(구 응용약물학회지) Vol.31 No.2

Among 14 subtypes of serotonin receptors (5-HTRs), 5-HT_2AR plays important roles in drug addiction and various psychiatric disorders. Agonists for 5-HT_2AR have been classified into three structural groups: phenethylamines, tryptamines, and ergolines. In this study, the structure-activity relationship (SAR) of phenethylamine and tryptamine derivatives for binding 5-HT_2AR was determined. In addition, functional and regulatory evaluation of selected compounds was conducted for extracellular signal-regulated kinases (ERKs) and receptor endocytosis. SAR studies showed that phenethylamines possessed higher affinity to 5-HT_2AR than tryptamines. In phenethylamines, two phenyl groups were attached to the carbon and nitrogen (R³) atoms of ethylamine, the backbone of phenethylamines. Alkyl or halogen groups on the phenyl ring attached to the β carbon exerted positive effects on the binding affinity when they were at para positions. Oxygen-containing groups attached to R³ exerted mixed influences depending on the position of their attachment. In tryptamine derivatives, tryptamine group was attached to the β carbon of ethylamine, and ally groups were attached to the nitrogen atom. Oxygen-containing substituents on large ring and alkyl substituents on the small ring of tryptamine groups exerted positive and negative influence on the affinity for 5-HT_2AR, respectively. Ally groups attached to the nitrogen atom of ethylamine exerted negative influences. Functional and regulatory activities of the tested compounds correlated with their affinity for 5-HT_2AR, suggesting their agonistic nature. In conclusion, this study provides information for designing novel ligands for 5-HT_2AR, which can be used to control psychiatric disorders and drug abuse.

Facilitation of serotonin-induced contraction of rat mesenteric artery by ketamine

Sang Woong Park,Hyun Ju Noh,Jung Min Kim,Bokyung Kim,Sung-Il Cho,Yoon Soo Kim,Nam Sik Woo,Sung Hun Kim,Young Min Bae 대한생리학회-대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.6

Ketamine is an anesthetic with hypertensive effects, which make it useful for patients at risk of shock. However, previous <i>ex vivo</i> studies reported vasodilatory actions of ketamine in isolated arteries. In this study, we reexamined the effects of ketamine on arterial tones in the presence and absence of physiological concentrations of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) by measuring the isometric tension of endothelium-denuded rat mesenteric arterial rings. Ketamine little affected the resting tone of control mesenteric arterial rings, but, in the presence of 5-HT (100~200 nM), ketamine (10~100 μM) markedly contracted the arterial rings. Ketamine did not contract arterial rings in the presence of NE (10 nM), indicating that the vasoconstrictive action of ketamine is 5-HT-dependent. The concentration-response curves (CRCs) of 5-HT were clearly shifted to the left in the presence of ketamine (30 μM), whereas the CRCs of NE were little affected by ketamine. The left shift of the 5-HT CRCs caused by ketamine was reversed with ketanserin, a competitive 5-HT_2Areceptor inhibitor, indicating that ketamine facilitated the activation of 5-HT_2A receptors. Anpirtoline and BW723C86, selective agonists of 5-HT_1B and 5-HT_2B receptors, respectively, did not contract arterial rings in the absence or presence of ketamine. These results indicate that ketamine specifically enhances 5-HT_2A receptor-mediated vasoconstriction and that it is vasoconstrictive in a clinical setting. The facilitative action of ketamine on 5-HT_2A receptors should be considered in ketamine-induced hypertension as well as in the pathogenesis of diseases such as schizophrenia, wherein experimental animal models are frequently generated using ketamine.

Facilitation of serotonin-induced contraction of rat mesenteric artery by ketamine

Park, Sang Woong,Noh, Hyun Ju,Kim, Jung Min,Kim, Bokyung,Cho, Sung-Il,Kim, Yoon Soo,Woo, Nam Sik,Kim, Sung Hun,Bae, Young Min The Korean Society of Pharmacology 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.6

Ketamine is an anesthetic with hypertensive effects, which make it useful for patients at risk of shock. However, previous ex vivo studies reported vasodilatory actions of ketamine in isolated arteries. In this study, we reexamined the effects of ketamine on arterial tones in the presence and absence of physiological concentrations of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) by measuring the isometric tension of endothelium-denuded rat mesenteric arterial rings. Ketamine little affected the resting tone of control mesenteric arterial rings, but, in the presence of 5-HT (100~200 nM), ketamine ($10{\sim}100{\mu}M$) markedly contracted the arterial rings. Ketamine did not contract arterial rings in the presence of NE (10 nM), indicating that the vasoconstrictive action of ketamine is 5-HT-dependent. The concentration-response curves (CRCs) of 5-HT were clearly shifted to the left in the presence of ketamine ($30{\mu}M$), whereas the CRCs of NE were little affected by ketamine. The left shift of the 5-HT CRCs caused by ketamine was reversed with ketanserin, a competitive 5-$HT_{2A}$ receptor inhibitor, indicating that ketamine facilitated the activation of 5-$HT_{2A}$ receptors. Anpirtoline and BW723C86, selective agonists of 5-$HT_{1B}$ and 5-$HT_{2B}$ receptors, respectively, did not contract arterial rings in the absence or presence of ketamine. These results indicate that ketamine specifically enhances 5-$HT_{2A}$ receptor-mediated vasoconstriction and that it is vasoconstrictive in a clinical setting. The facilitative action of ketamine on 5-$HT_{2A}$ receptors should be considered in ketamine-induced hypertension as well as in the pathogenesis of diseases such as schizophrenia, wherein experimental animal models are frequently generated using ketamine.