人胎盘滋养层细胞雌激素α,β受体免疫细胞化学研究

目的:探讨雌激素-α,β型受体（estrogen recoptor α,β）在人胎盘滋养层细胞的表达和定位。方法:采用细胞培养结合免疫细胞化学ABC法。结果:人胎盘及培养的人胎盘绒毛合体、细胞滋养层细胞均呈雌激素α,β受体免疫反应性,雌激素受体免疫反应阳性物质定位于胞核内,细胞质为阴性。结论:人胎盘滋养层细胞存在雌激素α,β受体,这为研究雌激素对人胎盘滋养层细胞的功能调控提供了形态学依据。     

人早期胎盘绒毛运动神经诱向因子1及其受体的免疫组织化学定位

为了解人胎盘绒毛是不存在运动神经诱向因子及其可能的功能意义，本实验用ＭＮＴＦ１单克隆抗体及抗独特型单克隆抗体在人早期胎盘绒毛石蜡切片上进行免疫组织化学反应，对人早期胎盘绒毛的ＭＮＴＦ１及其受体进行定位。结果显示，胎盘绒毛的细胞滋养层细胞，合体滋养层细胞和基质细胞均呈ＭＮＴＦ１强免疫反尖，ＭＮＴＦ１样免疫反应物质分布在胞质内，胞核内阴性。     

原癌基因c-fos、p53及运动神经诱向因子1受体在人早期胎盘绒毛的免疫组织化学定位

为了解原癌基因对胎盘滋养层细胞的影响，用原癌基因ｃ－ｆｏｓ、ｐ５３蛋白的抗体和运动神经诱向因子１（ｍｏｔｏｎｅｕｒｏｎｏｔｒｏｐｈｉｃｆａｃｔｏｒ１，ＭＮＴＦ１）抗独特型抗体的免疫组织化学技术，研究了ｃ－ｆｏｓ、ｐ５３及ＭＮＴＦ１受体在人早期胎盘的分布。人早期胎盘的两类滋养层细胞及绒毛中轴的基质细胞均呈ｃ－ｆｏｓ、ｐ５３免疫反应性，反应产物大部分布于胞核内。在相邻切片上，ｃ－ｆｏｓ免疫反应性细胞同样显示ＭＮＴＦ１受体免疫反应性，其反应物质分布于胞质内。以上结果提示，原癌基因ｃ－ｆｏｓ和ｐ５３可能参与调控绒毛细胞的增殖分化，而且ｃ－ｆｏｓ和ＭＮＴＦ１受体可能有协同调控作用。     

原癌基因c—fos,p53及运动神经诱向因子1受体在人早期胎盘绒…

为了解原癌基因对胎盘滋养层细胞的影响，用原癌基因ｃ－ｆｏｓ、ｐ５３蛋白的抗体和运动神经诱向因子抗独特特异抗体的免疫组织化学技术，研究了ｃ－ｆｏｓ、ｐ５３及ＭＦＴＦ１受体在人早期胎盘的分布。人早期胎盘的两类滋养层主绒毛中轴的基质细胞均呈ｃ－ｆｏｓ、ｐ５３免疫反应性，反应产物大部分布于胞核内，在相邻切片上，ｃ－ｆｏｓ免疫反应性细胞同样显示ＭＮＴＦ１受体免疫反应性，其反应物质分布于胞质内。以上结果提示，     

人胎盘蜕膜细胞5—羟色胺及其受体的免疫组织化学及原位杂？…

目的 观察５－羟色胺（５－ＨＴ）及其受体在人胎盘的定位及其含量的周龄变化。方法 采用免疫组织化学、原位杂交及图像分析技术。结果 人胎盘蜕膜细胞呈５－ＨＴ及其受体免疫反庆产物的相对含量随妊娠周龄的增加而增加。结论 人胎盘蜕膜细胞既能产生５－ＨＴ,又能表达５－ＨＴ受体。说明５－ＨＴ对蜕膜细胞可能具有自调节作用。     

大鼠实验性胃溃疡自愈其间胰岛β—EP、5—HT、5—HTR细胞的免疫组织化学研究

目的：探讨大鼠实验性胃溃疡自愈期间，胰岛β－内啡肽（β－EP）、5－羟色胺（5－HT）、5－羟色胺受体蛋白（5－HT）免疫反应细胞的变化。方法：免疫组织化学技术。结果：溃疡术后4d、10d胰岛，β－EP、5－HT、5－HTR细胞面数密度增高。β－EP、5－HT细胞于4d达高峰；5－HTR细胞于10d达高峰。邻片示；部分β－EP、5－HT及5－HTR阳性物质分别位于A或B细胞。结论：胰岛β－EP、5－HT、5－HTR细胞可能间接或直接参与大鼠实验性胃溃疡修复的调节。     

松驰素受体LGR7在人妊娠绒毛及胎盘组织中表达的研究

目的探讨松驰素受体LGR7在妊娠过程中的作用。方法应用免疫组织化学（免疫组化）链霉素抗生物素一过氧化物酶法和RT—PCR技术，对56例妊娠绒毛和胎盘组织中的LGR7检测。结果在早期妊娠的绒毛中，LGR7免疫反应呈现强阳性。主要分布于绒毛滋养层细胞的细胞膜上，在细胞滋养层细胞、合体滋养层细胞及滋养层细胞柱中都有较强的信号，并可见细胞滋养细胞的阳性反应明显强于合体滋养细胞；中、晚期妊娠的胎盘组织中LGR7呈现为强阳性或阳性，主要定位于合体滋养细胞的细胞膜上及毛细血管内皮细胞。免疫组化结果中，早期妊娠绒毛LGR7的强阳性率为90％，明显高于中晚期胎盘中的76．5％和73．7％，中、晚期妊娠分别与早期妊娠比较，差异均有统计学意义（P〈0．01）；但中期妊娠及晚期妊娠相比较，差异无统计学意义（P〉0．05）；早期妊娠的绒毛中LGR7mRNA的表达量为0．967±0．019，明显高于中晚期妊娠胎盘中的0．522±0．071和0．482±0．094，中、晚期妊娠分别与早期妊娠比较，差异均有统计学意义（P〈0．01）；中期妊娠及晚期妊娠比较，差异无统计学意义（P〉0．05）。结论LGR7可能是与妊娠早期滋养细胞侵袭有关的重要的因子，在妊娠的中晚期参与妊娠的维持。     

TNF-相关诱导凋亡因子及其受体在人胎盘组织中的免疫组织化学定位

目的 探讨TNF相关诱导凋亡因子(TRAIL)及其DR4、DR5受体在人胎盘组织的表达及其意义。方法 免疫组织化学结合图像分析定量方法观察TRAIL及其DR4、DR5受体在人胎盘组织的表达及其含量的周龄变化。结果 人胎盘滋养层细胞、绒毛基质细胞及毛细血管的内皮细胞均呈TRAIL及DR4、DR5受体免疫反应阳性，阳性反应物分布于胞膜及胞质，胞核阴性。在人胎盘绒毛的不同发育阶段中，TRAIL的含量相对稳定，而其受体DR4、DR5的含量随着周龄的增加而升高。结论 结果显示胎盘不仅能产生TRAIL，而且也是TRAIL的靶器官，TRAIL及其DR4、DR5受体系统可能参与胎盘的特免调节。     

原癌基因c-fos在人早期胎盘绒毛的分布及周龄变化

原癌基因ｃ－ｆｏｓ在人早期胎盘绒毛的分布及周龄变化新宇黄威权孙岚（第四军医大学）本文用免疫组织化学ＡＢＣ法，研究了原癌基因ｃ－ｆｏｓ在人早期胎盘绒毛的分布及周龄变化。免疫组织化学法显示：胎盘绒毛的细胞滋养层细胞、合体滋养层细胞和基质细胞均呈ｃ－ｆｏ...     

运动神经诱向因子1及其受体在人早期胎盘绒毛的分布及周龄变化

运动神经诱向因子１及其受体在人早期胎盘绒毛的分布及周龄变化新宇黄威权孙岚（第四军医大学）本文用免疫组织化学ＡＢＣ法，研究了运动神经诱向因子１（ＭＮＴＦ１）及其受体在人早期胎盘绒毛的分布及周龄变化。邻片免疫组织化学双标记法显示：胎盘绒毛的细胞滋养层细...     

豚鼠胃和小肠5—羟色胺及其受体免疫组织化学双标记研究

实验用邻片免疫组织化学双标记法，对豚鼠胃和小肠５－羟色胺（５－ＨＴ）及５－ＨＴ受体进行双标记研究。结果显示，胃底腺壁细胞，小肠的绒毛上皮细胞和小肠腺上皮细胞均呈５－ＨＴ受体阳性，５－ＨＴ阳性内分泌细胞散在分布于５－ＨＴ受体阳性的细胞间，一些５－ＨＴ阳性的内分泌细胞同样呈５－ＨＴ受体阳性。胃及小肠肌层的平滑肌细胞呈５－ＨＴ阴性反应但呈５－ＨＴ受体阳性反应，肌间神经丛的神经纤维呈５－ＨＴ阳性，神经元胞     

Serotoninergic receptors on human airway epithelial cells

There is accumulating evidence that points to a role of serotonin (5-hydroxytryptamine [5-HT]) in the pathophysiology of asthma. Therefore, we analyzed the expression of serotoninergic receptors (5-HTR), its linkage to intracellular calcium homeostasis, and its influence on the production and secretion of IL-6, prostaglandin E(2), the CCL-Chemokine CCL5/Rantes, and the CXC-chemokines CXCL8/IL-8, CXCL9/MIG, CXCL10/IP-10, and CXCL11/I-TAC in primary alveolar epithelial cells type II and the human lung cell lines A549 and BEAS-2B. Employing a PCR approach we were able to demonstrate mRNA expression of several 5-HTR, such as the heptahelical receptors 5-HTR1A, 5-HTR1B, 5-HTR1E, 5-HTR1F, 5-HTR2A, 5-HTR4, 5-HTR6, and 5-HTR7, as well as the ligand-gated ion channel 5-HTR3 in alveolar epithelial cells type II (AEC-II), A549, and BEAS-2B cells. To verify functional expression of 5-HTR subtypes, Ca(2+)-transients were analyzed. This enabled us to show that 5-HT induced an increase in intracellular calcium. Further experiments with isotype-selective receptor agonists allowed us to demonstrate that 5-HT induced calcium transients via activation of 5-HTR1, 5-HTR2, and 5-HTR3 in A549 and BEAS-2B cells. Moreover, we revealed that stimulation of 5-HTR1 and 5-HTR2 induced Ca(2+) mobilization from intracellular stores, whereas activation of 5-HTR3 induced Ca(2+) influx from the extracellular space. Functional studies indicated that activation of 5-HTR1B, 5-HTR1E/F, 5-HTR2, 5-HTR3, 5-HTR4, and 5-HTR7 regulated the release of the cytokine IL-6 and the CXC-chemokine CXCL8/IL-8. Our study shows that 5-HT stimulates different signaling pathways and regulates cytokine release in airway epithelial cells. In summary, our data implicate a pathophysiologic role of 5-HT in the asthmatic inflammatory responses in human airway epithelial cells.     

5-羟色胺及其受体在大鼠舌下腺的免疫组织化学研究

用免疫组织化学ABC法研究了大鼠舌下腺中5-羟色胺及其受体的分布情况,结果表明大鼠舌下腺的混合性腺泡的浆液性腺细胞、纹状管的上皮细胞均呈5-羟色胺免疫反应阳性.邻片双标染色发现5-羟色胺免疫反应阳性细胞也呈5-羟色胺受体免疫反应阳性,提示大鼠舌下腺有自分泌5-羟色胺之功能.     

大鼠实验性胃溃疡自愈期间胰岛β-EP、5-HT、5-HTR细胞的免疫组织化学研究

目的　探讨大鼠实验性胃溃疡自愈期间 ,胰岛 β-内啡肽 (β- EP)、5 -羟色胺 (5 - HT)、5 -羟色胺受体蛋白(5 - HTR)免疫反应细胞的变化。　方法　免疫组织化学技术。　结果　溃疡术后 4d、10 d胰岛 β- EP、5 - HT、5 - HTR细胞面数密度增高。β- EP、5 - HT细胞于 4d达高峰 ;5 - HTR细胞于 10 d达高峰。邻片示 :部分 β- EP、5 - HT及 5 - HTR阳性物质分别位于 A或 B细胞。　结论　胰岛 β- EP、5 - HT、5 - HTR细胞可能间接或直接参与大鼠实验性胃溃疡修复的调节。     

Serotonergic modulation of startle-escape plasticity in an African cichlid fish: a single-cell molecular and physiological analysis of a vital neural circuit

Social life affects brain function at all levels, including gene expression, neurochemical balance, and neural circuits. We have previously shown that in the cichlid fish Astatotilapia burtoni brightly colored, socially dominant (DOM) males face a trade-off between reproductive opportunities and increased predation risk. Compared with camouflaged subordinate (SUB) males, DOMs exposed to a loud sound pip display higher startle responsiveness and increased excitability of the Mauthner cell (M-cell) circuit that governs this behavior. Using behavioral tests, intracellular recordings, and single-cell molecular analysis, we show here that serotonin (5-HT) modulates this socially regulated plasticity via the 5-HT receptor subtype 2 (5-HTR(2)). Specifically, SUBs display increased sensitivity to pharmacological manipulation of 5-HTR(2) compared with DOMs in both startle-escape behavior and electrophysiological properties of the M-cell. Immunohistochemistry showed serotonergic varicosities around the M-cells, further suggesting that 5-HT impinges directly onto the startle-escape circuitry. To determine whether the effects of 5-HTR(2) are pre- or postsynaptic, and whether other 5-HTR subtypes are involved, we harvested the mRNA from single M-cells via cytoplasmic aspiration and found that 5-HTR subtypes 5A and 6 are expressed in the M-cell. 5-HTR(2), however, was absent, suggesting that it affects M-cell excitability through a presynaptic mechanism. These results are consistent with a role for 5-HT in modulating startle plasticity and increase our understanding of the neural and molecular basis of a trade-off between reproduction and predation.     

5-HTR2A基因启动子区-1438G/A多态性与抗精神病药源性肥胖的核心家系关联研究

目的探讨中国汉族首发精神分裂症(schizophrenia,SCH)患者抗精神病药物(antipsychoticagents,APS)治疗过程中体重增加是否与五羟色胺2A受体(5-hydroxytryptamine2Areceptor,5-HTR2A)基因启动区-1438G/A多态性相关。方法对84例首发精神分裂症患者(包含完整核心家系70个)APS(氯丙嗪或利培酮)单药治疗10周,治疗前后测量体重并计算体重指数。采用聚合酶链反应-限制性片段长度多态技术分析5-HTR2A基因启动区-1438G/A多态性基因型和等位基因分布频率,进行APS所致体重增加与5-HTR2A基因启动区-1438G/A多态性的相关分析、传递不平衡检验及数量性状传递不平衡检验。结果治疗10周后患者体重较基础体重增加(8.00±6.13)%。APS治疗10周后,体重增加≥7%和<7%患者组间,5-HTR2A基因-1438G/A多态性各基因型和等位基因分布频率差异均无统计学意义(P>0.05)。5-HTR2A基因-1438G/A多态性的各基因型之间各项指标的差异均无统计学意义(P>0.05);同时未发现5-HTR2A基因-1438G/A在不同体重增加组间存在传递不平衡。结论5-HTR2A基因-1438G/A多态性可能不是影响APS所致体重增加的主要遗传因素。     

Ionotropic 5-HT3 receptor agonist-induced motor responses in the hindlimbs of paraplegic mice

Centrally expressed 5-HT3 receptors (5-HTR3) are well known for their role in wakefulness, cognition, and nociception. However, clear evidence of their participation in motor control is still lacking despite specific 5-HTR3 expression in hindlimb motor areas of the spinal cord (i.e., lumbar laminae VII-IX). Here, we studied the acute effects of 4-amino-(6-chloro-2-pyridyl)-1-piperidine hydrochloride (SR 57227A), a potent and selective 5-HTR3 agonist, on hindlimb movement generation in complete paraplegic mice. The induced movements were assessed in open-field, air-stepping, and treadmill conditions using a combination of qualitative and quantitative methods. The results revealed that SR 57227A (1-4 mg/kg ip) produced hindlimb movements corresponding to scores ranging from 1 to 5 on the motor scales of Basso, Beattie, and Bresnahan and of Antri, Orsal, and Barthe. Additional analyses revealed that one-third of the movements displayed on a treadmill were "locomotor-like" (i.e., bilateral alternation), whereas only nonlocomotor movements were observed in the other testing conditions suggesting a task-dependent contribution of peripheral afferent inputs to these effects. Locomotor-like movements could also be induced in open field and air stepping if SR 57227A was combined with subthreshold doses of 5-carboxytryptamine (5-HT1A/7 receptor agonist), suggesting synergistic actions of these drugs on central neurons. These results demonstrate that 5-HTR3 activation can induce motor activity and, under some conditions, rhythmic locomotor-like movements in the hindlimbs of paraplegic mice providing evidence of an unsuspected role for this receptor subtype in hindlimb motor control.     

Effect of d-serine on the serotonin receptors of human platelets

Recent literature and our previous observations indicated the presence of both NMDA and serotonin type 3 receptors in human platelets with very similar ionic currents to that of cultured mammalian neuronal receptors. Baseline electrophysiological data shows similar profile for platelets from both normal and schizophrenic subjects, whereas serotonin receptor studies exhibited the presence of 5-hydroxytryptamine type-3 (5-HT3) currents in both normal and schizophrenic platelets significantly different from each other. The two major differences observed were: first, 5-HT3 receptors present in the platelets of schizophrenic patients were four times more sensitive to serotonin than those present in the platelets of normal subjects and, second, that d-serine in micro molar concentrations dampens this effect in platelets from schizophrenic patients but increases the sensitivity of serotonin for platelet 5-HT3 receptors of normal subjects. Patch clamp technique was used to measure the whole cell currents passing through serotonin receptors in these two types of human platelets. The currents were found to be 5-HT3 receptor currents as they were abolished by 10 μM d-tubocurarine. Similarly, micromolar concentrations of d-serine increased the sensitivity of 5HT3 receptor currents in the normal human platelets but decreased it in the platelets of the schizophrenic patients. This effect was reversed when d-amino acid oxidase (0.3 μM) was co applied with 100 μM of d-serine, raising the possibility that d-serine by itself may act as a modulator for platelet 5-HT3 receptor channel currents. These observations raised exciting new questions about the role of platelet serotonin receptors and their regulation by d-serine.