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1.
Semiconductor quantum dots (QDs) are nanometer-sized fluorescent probes suitable for advanced biological imaging. We used QDs to track individual glycine receptors (GlyRs) and analyze their lateral dynamics in the neuronal membrane of living cells for periods ranging from milliseconds to minutes. We characterized multiple diffusion domains in relation to the synaptic, perisynaptic, or extrasynaptic GlyR localization. The entry of GlyRs into the synapse by diffusion was observed and further confirmed by electron microscopy imaging of QD-tagged receptors.  相似文献   

2.
Prostaglandin E2 (PGE2) is a crucial mediator of inflammatory pain sensitization. Here, we demonstrate that inhibition of a specific glycine receptor subtype (GlyR alpha3) by PGE2-induced receptor phosphorylation underlies central inflammatory pain sensitization. We show that GlyR alpha3 is distinctly expressed in superficial layers of the spinal cord dorsal horn. Mice deficient in GlyR alpha3 not only lack the inhibition of glycinergic neurotransmission by PGE2 seen in wild-type mice but also show a reduction in pain sensitization induced by spinal PGE2 injection or peripheral inflammation. Thus, GlyR alpha3 may provide a previously unrecognized molecular target in pain therapy.  相似文献   

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4.
In mammals different isoforms of myosin heavy chain are encoded by the members of a multigene family. The expression of each gene of this family is regulated in a tissue- and developmental stage-specific manner as well as by hormonal and various pathological stimuli. In this study the molecular basis of isoform switches induced in myosin heavy chain by thyroid hormone was investigated. The expression of the myosin heavy chain gene family was analyzed in seven different muscles of adult rats subjected to hypo- or hyperthyroidism with complementary DNA probes specific for six different myosin heavy chain genes. The results demonstrate that all six genes are responsive to thyroid hormone. More interestingly, the same myosin heavy chain gene can be regulated by thyroid hormone in highly different modes, even in opposite directions, depending on the tissue in which it is expressed. Furthermore, the skeletal embryonic and neonatal myosin heavy chain genes, so far considered specific to these two developmental stages, can be reinduced by hypothyroidism in specific adult muscles.  相似文献   

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6.
为探明磷酸二酯酶(phosphodiesterase,PDE)在犬中枢神经活动中的作用,采用反转录聚合酶链反应检测18个PDE亚型在犬中枢神经不同部位中(大脑、小脑、延髓和脊髓各段)的表达分布,以高效液相色谱法检测环核苷酸在酶反应前后的含量变化,计算PDE活性。结果显示:在所测18个PDE亚型中,犬中枢神经的8个测定部位均有PDE1A,PDE1B,PDE1C,PDE2A,PDE3A,PDE3B,PDE4A,PDE4B,PDE4C,PDE4D,PDE5A,PDE7A,PDE7B,PDE8A,PDE8B,PDE9A,PDE11A表达,PDE10A均呈阴性表达,不同部位之间表达量差异较大,其中PDE4在各部位均呈优势表达;8个测定部位cAMP-PDE活性均高于cGMP-PDE活性,且不同部位PDE活性存在差异,其中脑组织cAMP-PDE活性和cGMP-PDE活性均高于脊髓各段。结果提示PDE可能在由cAMP和cGMP介导的中枢神经经信号传导中发挥着重要作用。  相似文献   

7.
Neurons containing the enzyme aromatic-L-amino-acid decarboxylase (AADC) but lacking either tyrosine hydroxylase or serotonin were found in the spinal cord of neonatal and adult rats by light and electron microscopic immunocytochemistry. The majority of these neurons localized to area X of Rexed contact ependyma. Thus, spinal AADC neurons have the enzymatic capacity to catalyze directly the conversion of the amino acids tyrosine, tryptophan, or phenylalanine to their respective amines tyramine, tryptamine, or phenylethylamine. These amines normally present in the central nervous system may be of potential clinical significance as endogenous psychotomimetics.  相似文献   

8.
为揭示凋亡相关基因和c-kit基因在脊髓损伤(spinal cord injury,SCI)后的相互关系,利用RT-PCR和免疫组化技术,分别检测了caspase-3、bax、bcl-2、c-kit的mRNA和蛋白在小鼠脊髓损伤后的表达。结果显示:在SCI后4 h开始出现大量的神经元和胶质细胞凋亡,其中促凋亡因子表达增加,而抑制凋亡因子表达量减少。caspase-3的mRNA和蛋白的表达在SCI后开始增加,并在72 h时达到峰值,bax表达规律与之基本一致,但是其峰值出现在24 h。随着时间的变化,bcl-2与c-kit的表达均表现为升高—降低—升高的变化趋势,与caspase-3和bax的变化趋势相反,且其调控作用均发生在caspase-3之前。此外,SCI后c-kit基因也呈现抑制凋亡的作用。表明:损伤是导致脊髓发生凋亡的因素之一;bcl-2是凋亡抑制因子,与bax共同控制细胞凋亡,并直接作用于下游caspase-3的表达。  相似文献   

9.
尾加压素Ⅱ受体(UrotensinⅡreceptor,UT)是一种7次跨膜G蛋白偶联受体,是目前发现的具有最强收缩血管作用的尾加压素Ⅱ的特异性受体。斑马鱼中存在4种不同的UT-like基因(uts2r1,uts2r2,uts2r3和uts2r4),分别位于斑马鱼4条不同染色体上。这4种uts2r基因核苷酸序列各不相同,uts2r1位于3号染色体上,其与12号染色体上的uts2r2基因蛋白序列相似度最高为47%,与6号染色体上的uts2r3和16号染色体上的uts2r4蛋白相似度分别是41.6%和33%。通过设计特异引物检测每种uts2r基因在斑马鱼成鱼各组织中的表达情况,结果显示3号染色体上的uts2r1基因在肾脏中表达量最高,6号染色体上的uts2r2基因在脊髓中表达量最高,12号染色体上的uts2r3基因在心脏中表达量最高,而16号染色体上的uts2r4基因在背肌中表达量最高。  相似文献   

10.
【目的】克隆红鳍笛鲷(Lutjanus erythropterus)视杆蛋白(RH1)基因和长波敏感视蛋白(LWS)基因,并分析其在早期发育不同阶段和成鱼不同组织的表达规律,为探究笛鲷属鱼类适应从表层浮游逐步转为下层底栖光环境生活的过程提供理论基础。【方法】利用RACE克隆红鳍笛鲷RH1基因和LWS基因cDNA全长,利用生物信息学软件对2个基因序列及编码的氨基酸序列进行分析,并利用实时荧光定量PCR检测其在红鳍笛鲷6个胚胎发育时期(原肠下包1/2期、胚孔封闭期、视囊期、晶体出现期、心脏跳动期、孵化出膜期)和5个仔鱼发育时期(1、3、10、15和20 d),以及成鱼不同组织(脑脏、心脏、肝脏、肌肉、黑色皮肤、红色皮肤、胃和视网膜)的表达规律。【结果】红鳍笛鲷RH1基因和LWS基因cDNA全长分别为1723 bp和1302 bp,其中RH1基因具有465 bp的3'-UTR和196 bp的5'-UTR,开放阅读框(ORF)长度为1062 bp,编码353个氨基酸残基;LWS基因具有213 bp的3'-UTR和15 bp的5'-UTR,ORF长度为1074 bp,编码357个氨基酸残基。系统发育分析结果显示,2个视蛋白基因均先与鲈形目等鱼类聚为一支,再与鳉形目、鲽形目和鲤形目等聚为一支,最后再与陆生脊椎动物聚为一支。实时荧光定量PCR检测结果显示,RH1基因在孵化出膜后15和20 d的相对表达量显著高于其他时期(P<0.05,下同),LWS基因在孵化出膜后20 d的相对表达量显著高于其他时期;2个视蛋白基因在成鱼不同组织的表达模式相似,在视网膜上的相对表达量均显著高于其他组织。【结论】红鳍笛鲷RH1和LWS基因的表达具有组织特异性,且在早期发育阶段的表达水平与红鳍笛鲷的生活习性变化相关,表明红鳍笛鲷RH1和LWS基因表达与其生活光环境的变化密切呼应,在早期发育变态阶段的光线调节,以及适应黑暗环境等方面发挥重要作用。  相似文献   

11.
运用实时荧光定量PCR技术,检测 IGF2H19 印记基因在长白猪和蓝塘猪中的表达水平.测定了1日龄和180日龄2个阶段的基因在各组织中的转录表达水平,按照生长性状分组,对组间的表达差异进行了比较分析.分析不同日龄长白、蓝塘猪各组织中 IGF2H19 的表达量.结果显示,仔猪初生体质量大的组肝脏组织中 IGF2 基因表达水平显著高于初生体质量轻的组;180日龄猪皮脂厚的组脂肪组织的 IGF2 基因表达量高于皮脂薄的组;1日龄仔猪肝脏、肌肉和胃组织中 IGF2 表达量高于其他测定组织和180日龄的各组织;180日龄蓝塘猪肾脏组织中的 H19 表达量显著升高.  相似文献   

12.
Synaptic potentials recorded in cell cultures of nerve and muscle   总被引:11,自引:0,他引:11  
Initially dissociated spinal cord and muscle cells derived from chick embryos differentiate sufficiently in tissue culture to form functional synaptic contacts. Spontaneous and evoked potentials recorded with intracellular microelectrodes resemble synaptic responses of adult spinal cord and neuromuscular junctions.  相似文献   

13.
The long-term, chronic, paralysis resulting from spinal cord injury in the cat has been reversed by the use of an alpha 2-adrenergic receptor agonist, clonidine. Administration of this drug resulted in "normalization" of sensory-motor and autonomic dysfunctions. Preliminary studies of the clonidine in humans with traumatically injured spinal cord indicate that autonomic dysreflexia can be controlled and spasticity minimized. The data suggest that biochemical and pharmacologic manipulation of receptors may ameliorate paralysis following traumatic injury to the spinal cord as well as to the brain and brainstem.  相似文献   

14.
A new type of agonist-binding subunit of rat neuronal nicotinic acetylcholine receptors (nAChRs) was identified. Rat genomic DNA and complementary DNA encoding this subunit (alpha 2) were cloned and analyzed. Complementary DNA expression studies in Xenopus oocytes revealed that the injection of messenger RNAs (mRNAs) for alpha 2 and beta 2 (a neuronal nAChR subunit) led to the generation of a functional nAChR. In contrast to the other known neuronal nAChRs, the receptor produced by the injection of alpha 2 and beta 2 mRNAs was resistant to the alpha-neurotoxin Bgt3.1. In situ hybridization histochemistry showed that alpha 2 mRNA was expressed in a small number of regions, in contrast to the wide distribution of the other known agonist-binding subunits (alpha 3 and alpha 4) mRNAs. These results demonstrate that the alpha 2 subunit differs from other known agonist-binding alpha-subunits of nAChRs in its distribution in the brain and in its pharmacology.  相似文献   

15.
【目的】研究牦牛和犏牛Dmrt7基因编码区序列和编码蛋白的结构,以及在睾丸组织中mRNA及其蛋白表达水平,探讨Dmrt7与犏牛雄性不育的关系,为揭示犏牛雄性不育的分子机理提供依据。【方法】利用分子克隆技术获得牦牛和犏牛Dmrt7基因编码区序列,并采用生物信息学方法对该基因及其编码蛋白的功能位点和二级结构等方面进行了预测和分析;通过半定量PCR技术检测Dmrt7基因mRNA在牦牛各组织器官中的表达水平;利用实时荧光定量PCR技术检测牦牛和犏牛睾丸组织中Dmrt7基因mRNA表达水平;并通过western blotting检测牦牛和犏牛睾丸组织中Dmrt7蛋白的表达水平。【结果】牦牛和犏牛Dmrt7基因cDNA序列一致,包含一个长度为1 113 bp的开放阅读框,编码370个氨基酸,具有完整的DM功能域,二级结构主要以无规则卷曲、α螺旋和延伸链为主。在牦牛各组织器官中,Dmrt7基因mRNA仅在睾丸组织中特异性表达。牦牛睾丸组织中Dmrt7mRNA和蛋白的表达水平极显著高于犏牛(P<0.01)。【结论】牦牛睾丸组织中Dmrt7基因mRNA和蛋白表达水平明显高于犏牛,且Dmrt7蛋白表达水平与其mRNA表达水平相一致。  相似文献   

16.
【目的】克隆山羊Eda基因CDS区全序列,并对其进行序列分析,研究Eda基因在毛囊生长周期不同阶段皮肤组织中的表达规律。【方法】以太行黑山羊为研究对象,采集其毛囊生长休止期、生长期和退行期背部皮肤组织,采用RT-PCR技术克隆山羊Eda基因,通过在线软件Blastn进行基因cDNA序列分析,用SMART进行氨基酸序列分析,利用SWISS-MODEL软件进行蛋白质结构分析;以β-actin为看家基因,采用实时荧光定量PCR技术对 mRNA在毛囊生长不同时期皮肤组织中的表达规律进行分析。【结果】太行黑山羊Eda-A1基因CDS区全长1 176 bp,编码391个氨基酸;Eda-A2比Eda-A1少6个碱基(nt1161-1166),编码389个氨基酸;山羊Eda蛋白氨基酸序列相似性在不同物种间均大于90%。SMART分析表明,CDS编码的蛋白质包含了TNF、跨膜区以及胶原等结构域。SWISS-MODEL软件预测结果表明,Eda-A2与Eda-A1在蛋白质表面结构上存在明显差异。毛囊退行期的皮肤组织中,Eda mRNA表达量最高,且极显著高于毛囊休止期和毛囊生长期(P<0.01),毛囊生长期的表达量中等且显著高于休止期(P<0.05),毛囊休止期的表达量较低。【结论】Eda基因CDS区在物种间保守性较强,Eda mRNA在毛囊生长周期不同阶段表达量有差异,推测Eda基因对毛囊生长周期的调控有一定影响。  相似文献   

17.
Binding of antibodies to effector cells by way of receptors to their constant regions (Fc receptors) is central to the pathway that leads to clearance of antigens by the immune system. The structure and function of this important class of receptors on immune cells is addressed through the molecular characterization of Fc receptors (FcR) specific for the murine immunoglobulin G isotype. Structural diversity is encoded by two genes that by alternative splicing result in expression of molecules with highly conserved extracellular domains and different transmembrane and intracytoplasmic domains. The proteins encoded by these genes are members of the immunoglobulin supergene family, most homologous to the major histocompatibility complex molecule E beta. Functional reconstitution of ligand binding by transfection of individual FcR genes demonstrates that the requirements for ligand binding are encoded in a single gene. These studies demonstrate the molecular basis for the functional heterogeneity of FcR's, accounting for the possible transduction of different signals in response to a single ligand.  相似文献   

18.
旨在分析并预测细粒棘球绦虫锌指蛋白(Echinococcus granulosus zinc finger peotein,Eg-ZFP)结构、功能及其在细粒棘球绦虫发育过程中的表达模式,预测其在细粒棘球绦虫防治中潜在的作用。通过对前期细粒棘球绦虫原头蚴(protoscolex,PSC)转录组高通量测序结果中Unigene进行筛选,克隆Eg-ZFP基因并进行序列分析,实时荧光定量PCR(Realtime fluorescence quantitative PCR,qPCR)分析Eg-ZFP在原头蚴及成虫中的表达特征,利用全量组织原位杂交(Whole mount in situ hybridization,WISH)检测该蛋白mRNA在原头蚴和成虫组织中的分布情况。结果显示,细粒棘球绦虫Eg-ZFP基因的CDS序列长852bp,具有完整的开放阅读框(852bp),编码283个氨基酸,预测表明Eg-ZFP分子质量大小及等电点为31.6ku和8.89;qPCR检测发现,Eg-ZFP在原头蚴及成虫阶段均有表达,且在成虫阶段表达量高;全量组织原位杂交检测结果表明,Eg-ZFP mRNA在原头蚴及成虫中分布广泛,且在成虫的生殖系统中丰度较高,提示Eg-ZFP与虫体生殖发育有关。结果表明Eg-ZFP在细粒棘球绦虫发育过程中具有重要作用,为进一步研究Eg-ZFP在E.granulosus防治中的作用奠定基础。  相似文献   

19.
杨静  杨军  黄继超  何玮玲  张驰  黄明 《中国农业科学》2012,45(19):4057-4066
【目的】研究食源性金黄色葡萄球菌各血清型肠毒素基因的分布,并进一步分析其时序性表达规律。【方法】针对51株各类食品中分离的金黄色葡萄球菌菌株,应用PCR技术对11种葡萄球菌肠毒素进行基因分型;提取细菌总RNA,以ftsZ和ropB作为内标基因,使用反转录荧光定量PCR研究各肠毒素基因在细菌生长周期中的表达水平变化。【结果】除see、ses和set外,其余8种肠毒素基因在51株食源性金黄色葡萄球菌菌株中均有检出,且sei和seg的检出率最高(27.45%)。各肠毒素基因在mRNA水平的时序性表达规律基本一致,均在对数后期达到峰值,随后快速下降。以内标基因为参照,同一菌株中不同肠毒素基因的相对表达量以及不同菌株中同一肠毒素基因的相对表达量均差异较大。【结论】本文系统研究了肠毒素基因在食源性金黄色葡萄球菌中的分布,并探究了肠毒素基因的时序性表达规律,对金黄色葡萄球菌毒力机制研究与食品质量安全控制具有参考意义。  相似文献   

20.
[目的]克隆小菜蛾表皮蛋白基因(Plutella xylostella cuticular protein,PxyICP),分析其序列特征和发育表达模式,为深入研究PxyICP在小菜蛾表皮形成中的生理功能提供科学参考.[方法]以小菜蛾为试验材料,采用反转录聚合酶链式反应(RT-PCR)和快速扩增cDNA末端(RACE)技术克隆PxyICP,以生物信息学方法对其序列特征进行分析,并采用实时荧光定量PCR研究PxyICP mRNA在小菜蛾不同发育时期中的相对表达量.[结果]克隆获得的PxyICP基因开放阅读框长531 bp,编码177个氨基酸,相对分子质量约18.90 kD,等电点为5.32.氨基酸序列分析结果表明,小菜蛾表皮蛋白第1~16位氨基酸是参与跨膜蛋白转移的信号肽,且该序列具有昆虫表皮蛋白CPR家族中RR-1型保守基序的典型特征.不同发育时期的实时荧光定量PCR分析结果表明,PxyICP在小菜蛾各发育时期的表达量不同,其中PxyICP在2、3和4龄幼虫、蛹和成虫中的表达量分别是1龄幼虫的2.43、0.51、0.63、3.19和12.32倍,以在成虫的表达量最高.[结论]成功克隆获得PxyICP基因,根据其发育表达模式推测PxyICP蛋白参与小菜蛾成虫表皮的硬化和黑化等生理过程.  相似文献   

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