目的:探讨中药复方制剂参松养心胶囊对KV1.4钾通道C型(KV1.4△N)失活的影响。方法:将Kv1.4ΔNmRNA注射入非洲爪蟾卵母细胞中,使用双电极钳制法(Two electrodes voltage clamp TEV)观察参松养心胶囊对KV1.4ΔN电生理特性的影响。结果:参...目的:探讨中药复方制剂参松养心胶囊对KV1.4钾通道C型(KV1.4△N)失活的影响。方法:将Kv1.4ΔNmRNA注射入非洲爪蟾卵母细胞中,使用双电极钳制法(Two electrodes voltage clamp TEV)观察参松养心胶囊对KV1.4ΔN电生理特性的影响。结果:参松养心胶囊对Kv1.4ΔN通道的峰电流有抑制作用,这种阻滞作用具有电压依赖性,随电位的升高而作用加强,符合单指数和线性关系。参松养心胶囊可加速KV1.4ΔN钾通道电流的失活过程,同时可使KV1.4ΔN通道失活后的恢复减慢。结论:参松养心胶囊能显著抑制KV1.4钾通道电流,这可能是其抗心律失常作用的机制之一。展开更多
Kinetics of the low threshold T-type Ca2+ channel is studied with single electrode voltage damp technique on brain slices of the cat lateral geniculate nucleus (LGN). Space damp is dramatically improved by blocking va...Kinetics of the low threshold T-type Ca2+ channel is studied with single electrode voltage damp technique on brain slices of the cat lateral geniculate nucleus (LGN). Space damp is dramatically improved by blocking various K+ and Na+ channels, decreasing Ca2+ current and selecting proper holding potentials. Results from this study are similar to those obtained from acutely dissociated LGN neurons of the rat, indicating that the kinetics of T-Ca2+ channels of the cat LGN neurons is the same as that of the rat LGN. The result reported previously on the cat LGN may result from a defect in space damp.展开更多
Pea aphid,Acyrthosiphon pisum,is a serious pest of many different leguminous plants,and it mainly relies on its odorant receptors(Ors)to discriminate among host species.However,less is known about the role that Ors pl...Pea aphid,Acyrthosiphon pisum,is a serious pest of many different leguminous plants,and it mainly relies on its odorant receptors(Ors)to discriminate among host species.However,less is known about the role that Ors play in the host plant location.In this study,we identified a novel conserved odorant receptor clade by phylogenetic analysis,and conducted the functional analysis of ApisOr23 in A.pisum.The results showed that the homologous Ors from A.pisum,Aphis glycines and Aphis gossypii share 94.28% identity in amino acid sequences.Moreover,conserved motifs were analyzed using the annotated homologous Or23 from eight aphid species,providing further proof of the high conservation level of the Or23 clade.According to the tissue expression pattern analysis,ApisOr23 was mainly expressed in the antennae.Further functional study using a heterologous Xenopus expression system revealed that ApisOr23 was tuned to five plant volatiles,namely trans-2-hexen-1-al,cis-2-hexen-1-ol,1-heptanol,4’-ethylacetophenone,and hexyl acetate.Among them,trans-2-hexen-1-al,which is one of the main volatile organic compounds released from legume plants,activated the highest response of ApisOr23.Our findings suggest that the conserved Or23 clade in most aphid species might play an important role in host plant detection.展开更多
Premating behaviors mediated by pheromones play pivotal roles in animal mating choices.In natural populations of the striped stem borer Chilo suppressalis and the rice leaf roller Cnaphalocrocis medinalis in the rice ...Premating behaviors mediated by pheromones play pivotal roles in animal mating choices.In natural populations of the striped stem borer Chilo suppressalis and the rice leaf roller Cnaphalocrocis medinalis in the rice field habitat,we discovered that Z11-16:Ald,a major component of the C.suppressalis pheromone,modulated the premating behavior of C.medinalis.Z11-16:Ald evoked a strong olfactory response in male antennae and strongly inhibited the sex pheromone trapping of male C.medinalis in the field.The functions of three C.medinalis sex pheromone receptor genes(CmedPR1–3)were verified through heterologous expression in Xenopus oocytes.CmedPR1 responded to Z11-18:OH and Z11-18:Ald,as well as the interspecific pheromone compound Z11-16:Ac of sympatric species;CmedPR2 responded to Z13-18:OH and Z13-18:Ald,as well as the sex pheromone compounds Z11-16:Ald and Z9-16:Ald of sympatric species;and CmedPR3 responded to Z11-18:OH and Z13-18:OH,as well as the interspecific pheromones Z11-16:OH,Z9-16:Ald,Z11-16:Ac,and Z11-16:Ald of sympatric species.Thus,CmedPR2 and CmedPR3 share the ligand Z11-16:Ald,which is not a component of the C.medinalis sex pheromone.Therefore,the sex pheromones of interspecific species affected the input of neural signals by stimulating the sex pheromone receptors on the antennae of male C.medinalis moths,thereby inhibiting the olfactory responses of the male moths to the sex pheromones.Our results demonstrate chemical communication among sympatric species in the rice field habitat,the recognition of intra-and interspecific sex pheromones by olfactory receptors,and how insect premating behaviors are modulated to possibly affect resource partitioning.展开更多
Ion channels,as membrane proteins,are the sensors of the cell.They act as the first line of communication with the world beyond the plasma membrane and transduce changes in the external and internal environments into ...Ion channels,as membrane proteins,are the sensors of the cell.They act as the first line of communication with the world beyond the plasma membrane and transduce changes in the external and internal environments into unique electrical signals to shape the responses of excitable cells.Because of their importance in cellular communication,ion channels have been intensively studied at the structural and functional levels.Here,we summarize the diverse approaches,including molecular and cellular,chemical,optical,biophysical,and computational,used to probe the structural and functional rearrangements that occur during channel activation（or sensitization）,inactivation（or desensitization）,and various forms of modulation.The emerging insights into the structure and function of ion channels by multidisciplinary approaches allow the development of new pharmacotherapies as well as new tools useful in controlling cellular activity.展开更多
The roles of voltage-dependent K^＋ channels during activation and damage in alveolar macrophages （AMs） exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage,...The roles of voltage-dependent K^＋ channels during activation and damage in alveolar macrophages （AMs） exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage, and were adjusted to 5× 10^5/mL. After AMs were exposed to different concentrations （0, 25, 50, 100, 200 μg/mL） of quartz particles and 100 μg/mL amorphous silica particles for 24 h, the voltage-depended K^＋ current in AMs was measured by using patch clamp technique. Meanwhile the leakage of lactate dehydrogenase （LDH） and the viability of AMs were detected respectively. Patch clamp studies demonstrated that AMs possessed outward delayed and inward rectifying K^＋ current. Exposure to quartz particles increased the outward delayed K^＋ current but it had no effect on inward rectifier K^＋ current in AMs. Neither of the two K^＋ channels in AMs was affected by amorphous silica particles. Cytotoxicity test showed that both silica particles could damage AM membrane and result in significant leakage of LDH （P〈0.05）. MTT studies, however, showed that only quartz particles reduced viability of AMs （P〈0.05）. It is concluded that quartz parti- cles can activate the outward delayed K^＋ channel in AMs, which may act as an activating signal in AMs to initiate an inflammatory response during damage and necrosis in AMs induced by exposure to quartz particle. K^＋ channels do not contribute to the membrane damage of AMs.展开更多
Transient receptor potential vanilloid 1(TRPV1)channel plays an important role in a wide range of physiological and pathological processes,and a comprehensive understanding of TRPV1 gating will create opportunities fo...Transient receptor potential vanilloid 1(TRPV1)channel plays an important role in a wide range of physiological and pathological processes,and a comprehensive understanding of TRPV1 gating will create opportunities for therapeutic intervention.Recent incredible advances in cryo-electron microscopy(cryo-EM)have yielded high-resolution structures of all TRPV subtypes(TRPV1-6)and all of them share highly conserved six transmembrane(TM)domains(S1-S6).As revealed by the open structures of TRPV1 in the presence of a bound vanilloid agonist(capsaicin or resiniferatoxin),TM helices S1 to S4 form a bundle that remains quiescent during channel activation,highlighting differences in the gating mechanism of TRPV1 and voltage-gated ion channels.Here,however,we argue that the structural dynamics rather than quiescence of S1-S4 domains is necessary for capsaicin-mediated activation of TRPV1.Using fluorescent unnatural amino acid(flUAA)incorporation and voltage-clamp fluorometry(VCF)analysis,we directly observed allostery of the S1-S4 bundle upon capsaicin binding.Covalent occupation of VCF-identified sites,single-channel recording,cell apoptosis analysis,and exploration of the role of PSFL828,a novel non-vanilloid agonist we identified,have collectively confirmed the essential role of this coordinated S1-S4 motility in capsaicin-mediated activation of TRPV1.This study concludes that,in contrast to cryo-EM structural studies,vanilloid agonists are also required for S1-S4 movement during TRPV1 activation.Redefining the gating process of vanilloid agonists and the discovery of new non-vanilloid agonists will allow the evaluation of new strategies aimed at the development of TRPV1 modulators.展开更多
文摘目的:探讨中药复方制剂参松养心胶囊对KV1.4钾通道C型(KV1.4△N)失活的影响。方法:将Kv1.4ΔNmRNA注射入非洲爪蟾卵母细胞中,使用双电极钳制法(Two electrodes voltage clamp TEV)观察参松养心胶囊对KV1.4ΔN电生理特性的影响。结果:参松养心胶囊对Kv1.4ΔN通道的峰电流有抑制作用,这种阻滞作用具有电压依赖性,随电位的升高而作用加强,符合单指数和线性关系。参松养心胶囊可加速KV1.4ΔN钾通道电流的失活过程,同时可使KV1.4ΔN通道失活后的恢复减慢。结论:参松养心胶囊能显著抑制KV1.4钾通道电流,这可能是其抗心律失常作用的机制之一。
文摘Kinetics of the low threshold T-type Ca2+ channel is studied with single electrode voltage damp technique on brain slices of the cat lateral geniculate nucleus (LGN). Space damp is dramatically improved by blocking various K+ and Na+ channels, decreasing Ca2+ current and selecting proper holding potentials. Results from this study are similar to those obtained from acutely dissociated LGN neurons of the rat, indicating that the kinetics of T-Ca2+ channels of the cat LGN neurons is the same as that of the rat LGN. The result reported previously on the cat LGN may result from a defect in space damp.
基金funded by the National Natural Science Foundation of China(31572072 and 31725023)the Intergovernmental International Science,Technology and Innovation Cooperation Key Project,China(2019YFE0105800)the Shenzhen Science and Technology Program,China(KQTD20180411143628272)。
文摘Pea aphid,Acyrthosiphon pisum,is a serious pest of many different leguminous plants,and it mainly relies on its odorant receptors(Ors)to discriminate among host species.However,less is known about the role that Ors play in the host plant location.In this study,we identified a novel conserved odorant receptor clade by phylogenetic analysis,and conducted the functional analysis of ApisOr23 in A.pisum.The results showed that the homologous Ors from A.pisum,Aphis glycines and Aphis gossypii share 94.28% identity in amino acid sequences.Moreover,conserved motifs were analyzed using the annotated homologous Or23 from eight aphid species,providing further proof of the high conservation level of the Or23 clade.According to the tissue expression pattern analysis,ApisOr23 was mainly expressed in the antennae.Further functional study using a heterologous Xenopus expression system revealed that ApisOr23 was tuned to five plant volatiles,namely trans-2-hexen-1-al,cis-2-hexen-1-ol,1-heptanol,4’-ethylacetophenone,and hexyl acetate.Among them,trans-2-hexen-1-al,which is one of the main volatile organic compounds released from legume plants,activated the highest response of ApisOr23.Our findings suggest that the conserved Or23 clade in most aphid species might play an important role in host plant detection.
基金supported financially by NSF of China(Grant No.31871956)Zhejiang Province Key R&D Program(Grant No.2018C02027)to Y.D.
文摘Premating behaviors mediated by pheromones play pivotal roles in animal mating choices.In natural populations of the striped stem borer Chilo suppressalis and the rice leaf roller Cnaphalocrocis medinalis in the rice field habitat,we discovered that Z11-16:Ald,a major component of the C.suppressalis pheromone,modulated the premating behavior of C.medinalis.Z11-16:Ald evoked a strong olfactory response in male antennae and strongly inhibited the sex pheromone trapping of male C.medinalis in the field.The functions of three C.medinalis sex pheromone receptor genes(CmedPR1–3)were verified through heterologous expression in Xenopus oocytes.CmedPR1 responded to Z11-18:OH and Z11-18:Ald,as well as the interspecific pheromone compound Z11-16:Ac of sympatric species;CmedPR2 responded to Z13-18:OH and Z13-18:Ald,as well as the sex pheromone compounds Z11-16:Ald and Z9-16:Ald of sympatric species;and CmedPR3 responded to Z11-18:OH and Z13-18:OH,as well as the interspecific pheromones Z11-16:OH,Z9-16:Ald,Z11-16:Ac,and Z11-16:Ald of sympatric species.Thus,CmedPR2 and CmedPR3 share the ligand Z11-16:Ald,which is not a component of the C.medinalis sex pheromone.Therefore,the sex pheromones of interspecific species affected the input of neural signals by stimulating the sex pheromone receptors on the antennae of male C.medinalis moths,thereby inhibiting the olfactory responses of the male moths to the sex pheromones.Our results demonstrate chemical communication among sympatric species in the rice field habitat,the recognition of intra-and interspecific sex pheromones by olfactory receptors,and how insect premating behaviors are modulated to possibly affect resource partitioning.
基金supported by grants from the National Natural Science Foundation of China(91132303,30830035)the National Basic Research Development Program of China (2011CBA00408)the China Postdoctoral Science Foundation(2012M511105)
文摘Ion channels,as membrane proteins,are the sensors of the cell.They act as the first line of communication with the world beyond the plasma membrane and transduce changes in the external and internal environments into unique electrical signals to shape the responses of excitable cells.Because of their importance in cellular communication,ion channels have been intensively studied at the structural and functional levels.Here,we summarize the diverse approaches,including molecular and cellular,chemical,optical,biophysical,and computational,used to probe the structural and functional rearrangements that occur during channel activation（or sensitization）,inactivation（or desensitization）,and various forms of modulation.The emerging insights into the structure and function of ion channels by multidisciplinary approaches allow the development of new pharmacotherapies as well as new tools useful in controlling cellular activity.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30671743)
文摘The roles of voltage-dependent K^＋ channels during activation and damage in alveolar macrophages （AMs） exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage, and were adjusted to 5× 10^5/mL. After AMs were exposed to different concentrations （0, 25, 50, 100, 200 μg/mL） of quartz particles and 100 μg/mL amorphous silica particles for 24 h, the voltage-depended K^＋ current in AMs was measured by using patch clamp technique. Meanwhile the leakage of lactate dehydrogenase （LDH） and the viability of AMs were detected respectively. Patch clamp studies demonstrated that AMs possessed outward delayed and inward rectifying K^＋ current. Exposure to quartz particles increased the outward delayed K^＋ current but it had no effect on inward rectifier K^＋ current in AMs. Neither of the two K^＋ channels in AMs was affected by amorphous silica particles. Cytotoxicity test showed that both silica particles could damage AM membrane and result in significant leakage of LDH （P〈0.05）. MTT studies, however, showed that only quartz particles reduced viability of AMs （P〈0.05）. It is concluded that quartz parti- cles can activate the outward delayed K^＋ channel in AMs, which may act as an activating signal in AMs to initiate an inflammatory response during damage and necrosis in AMs induced by exposure to quartz particle. K^＋ channels do not contribute to the membrane damage of AMs.
基金supported by the Natural Science Foundation of Jiangsu Province(BK20202002)the National Natural Science Foundation of China(81603409,31900808,81902480,21977021,31570832,31971146,and 31971042)+4 种基金Innovation and Entrepreneurship Talent Program of Jiangsu ProvinceState Key Laboratory of Utilization of Woody Oil Resource(2019XK2002)the Natural Science Foundation of Hunan Province(2018JJ1012)Hunan“Huxiang”High-level Talent Program(2021)“Xing Yao”Leading Scholars of China Pharmaceutical University(2021)。
文摘Transient receptor potential vanilloid 1(TRPV1)channel plays an important role in a wide range of physiological and pathological processes,and a comprehensive understanding of TRPV1 gating will create opportunities for therapeutic intervention.Recent incredible advances in cryo-electron microscopy(cryo-EM)have yielded high-resolution structures of all TRPV subtypes(TRPV1-6)and all of them share highly conserved six transmembrane(TM)domains(S1-S6).As revealed by the open structures of TRPV1 in the presence of a bound vanilloid agonist(capsaicin or resiniferatoxin),TM helices S1 to S4 form a bundle that remains quiescent during channel activation,highlighting differences in the gating mechanism of TRPV1 and voltage-gated ion channels.Here,however,we argue that the structural dynamics rather than quiescence of S1-S4 domains is necessary for capsaicin-mediated activation of TRPV1.Using fluorescent unnatural amino acid(flUAA)incorporation and voltage-clamp fluorometry(VCF)analysis,we directly observed allostery of the S1-S4 bundle upon capsaicin binding.Covalent occupation of VCF-identified sites,single-channel recording,cell apoptosis analysis,and exploration of the role of PSFL828,a novel non-vanilloid agonist we identified,have collectively confirmed the essential role of this coordinated S1-S4 motility in capsaicin-mediated activation of TRPV1.This study concludes that,in contrast to cryo-EM structural studies,vanilloid agonists are also required for S1-S4 movement during TRPV1 activation.Redefining the gating process of vanilloid agonists and the discovery of new non-vanilloid agonists will allow the evaluation of new strategies aimed at the development of TRPV1 modulators.
