胰岛淀粉样多肽和甲硫氨酸-脑啡肽在家兔和大鼠背根神经节细胞内共存的免疫组织化学研究

为进一步探讨背根神经节细胞间生物活性物质内的关系，用免疫组织化学ＡＢＣ法，观察到家兔和大鼠背根神经节细胞内有胰岛淀粉样多肽免疫反应。用相邻镜像邻片免疫双标记技术，证实甲硫氨酸－脑啡肽免疫反应的细胞内同时含有胰岛淀粉样多肽免疫反应物质。胰岛淀粉样多肽与甲硫氨酸－脑啡肽在哺乳类动物背根神经节细胞内的共存结果，为进一步探讨背根神经节细胞内生物活性物质间的关系提供了形态学证据，并为胰岛淀粉样多肽的胰腺外研究提供了新的资料。     

胰岛淀粉样多肽与5—羟色胺,多巴胺在大鼠和家兔背根神经节…

用免疫组织化学ＡＢＣ法，在大鼠和家兔背根神经节内观察到胰岛淀粉样多肽免疫阳性细胞，用相邻镜像邻片双得免疫反应技术，证实它分别与５－羟色胺和多巴胺在大鼠和家兔背根神经节细胞内共存，为胰岛淀粉样多肽的胰腺外研究提供新的资料，为进一步研究背根神经节细胞内生物活性物质间的关系提供形态学证据。     

胰岛淀粉样多肽和甲硫氨酸—脑啡肽在家兔和大鼠背根神经节细胞内共存的

为进一步探讨背根根神经节细胞间生物活性物质内的关系，用免疫组织化学ＡＢＣ法，观察到家兔和大鼠背根神经节细胞内有胰岛淀粉样多肽免疫反应。用相邻镜像邻片免疫双标主民的，证实甲硫氨酸－脑啡肽免疫反主尖的细胞内同时含有胰岛淀粉样多肽免疫反应物质。     

甘丙肽和经典神经递质在大鼠和家兔背根神经节细胞内共存的免疫组织化学研究

为研究背根神经节细胞内生物活性物质间的关系，用免疫组织化学ＡＢＣ法，证实大鼠和家兔背根神经节存在甘丙肽免疫阳性反应细胞。用相邻镜像邻片免疫双标记技术，观察到甘丙肽分别与５－ＨＴ和ＤＡ在家兔和大鼠背根神经节细胞内共存。此结果为进一步研究背根神经节细胞内生物活性物质间的关系提供了形态学资料。     

甘丙肽和经典神经递质在大鼠和家兔背根神经节细胞内共存的免疫组织…

为研究背根神经节细胞内生笺活性质间的关系，用免疫组织化学ＡＢＣ法，证实大鼠和家兔背根神经节存在甘丙肽免疫阳反应细胞。用相邻镜像邻片免疫双标记技术，观察到甘丙肽分别与５－ＨＴ和ＤＡ在家兔和大鼠背根神经节细胞内共存。此结果为进一步研究背根神经节细胞内生物活性质间的关系提供了形态学资料。     

胰岛淀粉样多肽与甘丙肽在家兔和大鼠脊神经节细胞内分布与共存…

用免疫组织化学ＡＢＣ法和相邻镜像切片免疫双标记技术，对胰岛淀粉样多肽与甘丙肽在家兔和大鼠脊神经节细胞内的分布与共存进行了观察。结果发现；在家兔和大鼠脊神经节存在胰岛淀粉样多肽与甘丙肽免疫反应阳性细胞。在相邻像切片上，胰岛淀粉样多肽免疫反应阳性细胞与甘丙肽免疫反应阳性细胞呈镜像式对应重叠，证实二者共存，胰岛淀粉样多肽与甘丙肽在脊神经节细胞内的共存，为进一步研究脊神经节细胞内生物活性物质间的关系提供了     

胰岛淀粉样多肽与甘丙肽在家兔和大鼠脊神经节细胞内分布与共存的免疫组织化学研究

用免疫组织化学ABC法和相邻镜像切片免疫双标记技术,对胰岛淀粉样多肽与甘丙肽在家免和大鼠脊神经节细胞内的分布与共存进行了观察.结果发现:在家兔和大鼠脊神经节存在胰岛淀粉样多肽与甘丙肽免疫反应阳性细胞.在相邻镜像切片上,胰岛淀粉样多肽免疫反应阳性细胞与甘丙肽免疫反应阳性细胞呈镜像式对应重叠,证实二者共存.胰岛淀粉样多肽与甘丙肽在脊神经节细胞内的共存,为进一步研究脊神经节细胞内生物活性物质间的关系提供了形态学证据,并为胰岛淀粉样多肽的胰腺外研究提供了新的资料.     

IAPP免疫反应细胞在成年大鼠胃肠胰系统的分布与形态学研究

用免疫组织化学和免疫电镜双标记法研究了胰淀粉样多肽免疫反应细胞在正常大鼠胃肠胰系统的分布、定位和形态特征，并观察了ＩＡＰＰ与胰岛素在胰腺的共存。结果表明，大鼠胰腺和胃道各段均见有ＩＡＰＰ免疫反应细胞。在胰腺，ＩＡＰＰ与胰岛素共存于胰岛的Ｂ细胞内，也共存在散在于外分泌部的胰岛素免疫反应细胞内。胃肠道粘膜内ＩＡＰＰ免疫反应细胞主要存在于上皮和固有膜，它们的数量以胃和十二指肠较多，结肠较少。本研究首次发     

甘丙肽和经典递质在大鼠和家兔背根节细胞内共存的免疫组织化学研究

甘丙肽和经典递质在大鼠和家兔背根节细胞内共存的免疫组织化学研究（解放军北京医学高等专科学校解剖学教研室，北京１０００７１王雪岷，周长满，单文戈，鄂玲玲）我们曾用免疫组织化学ＰＡＰ法证明经典神经递质多巴胺（ＤＡ）和５－ＨＴ在鸡背根节细胞的分布和其表达的...     

MEF2C调控大鼠脊髓背根节感觉神经元P物质和低分子量神经丝微管蛋白的表达

目的:研究肌细胞增强因子2C(MEF2C)在大鼠脊髓背根神经节神经元细胞内的表达及其与P物质和低分子量神经丝微管蛋白合成的关系。方法:分离培养背根神经节神经元细胞,然后暴露于不同浓度的神经生长因子下24h,最后用实时聚合酶链反应技术检测P物质与低分子量神经丝微管蛋白基因在背根神经节神经元细胞内的表达。通过化学转染方法将合成的3条siRNA-MEF2C分别转染PC12细胞株,并用实时聚合酶链反应技术筛选干扰效率最高的siRNA。采用化学转染方法干扰背根神经节神经元细胞内MEF2C的表达,在高浓度神经生长因子刺激背根神经节神经元细胞后采用实时聚合酶链反应技术检测干扰后背根神经节神经元细胞内P物质与低分子量神经丝微管蛋白基因的表达。结果:P物质及低分子量神经丝微管蛋白基因表达随刺激用神经生长因子浓度的升高而升高。使用化学转染方法成功地干扰背根神经节神经元细胞内MEF2C的表达,MEF2C较对照组下降52%,同时没有检测到对cAMP反应元件结合蛋白表达的影响。实验组背根神经节神经元细胞内P物质在RNA水平较对照组下降了39%,而低分子量神经丝微管蛋白在RNA水平较对照组下降了62%。结论:神经生长因子促进大鼠脊髓背根节感觉神经元内P物质与低分子量神经丝微管蛋白的合成。大鼠背根神经节神经元细胞内P物质及低分子量神经丝微管蛋白基因表达受MEF2C调控。     

胚胎及新生期大鼠胃肠道胰岛淀粉样多肽与生长抑素的免疫组织化学定位

为探讨胚胎期及新生期大鼠胃肠道的胰岛淀粉样多肽（ＩＡＰＰ）与其他生物活性物质的关系，用免疫组织化学ＰＡＰ法，在相邻切片上分别显示第１５～２１ｄ大鼠胚胎及新生大鼠胃肠道的ＩＡＰＰ免疫反应性（ＩＲ）细胞和生长抑素（ＳＳ）ＩＲ细胞，观察了ＩＡＰＰ与ＳＳ在胚胎及新生大鼠胃肠道的定位分布。结果表明：第１５ｄ胚胎，胃肠道内未见ＩＡＰＰ－和ＳＳ－ＩＲ细胞。第１７ｄ胚胎，ＩＡＰＰ－与ＳＳ－ＩＲ细胞均很少，免疫染色较弱，着色较浅，免疫反应性细胞散布于分化未完全的肠上皮细胞间。胚胎１９ｄ，ＩＡＰＰ－和ＳＳ－ＩＲ细胞出现于胃和小肠，以十二指肠为多。胚胎第２１ｄ及新生期，胃肠道各段均可见到ＩＡＰＰ－和ＳＳ－ＩＲ细胞。经邻片比较证明，胚胎第１９ｄ始至新生期，十二指肠和空肠中部分ＩＡＰＰ－ＩＲ细胞与部分ＳＳ－ＩＲ细胞的定位相同，表明ＩＡＰＰ与ＳＳ在十二指肠和空肠Ｄ细胞中有共存。胃、回肠、结肠和直肠未见到ＩＡＰＰ与ＳＳ有细胞内共存的现象。本文对上述结果的意义进行了讨论     

The 5-HT2A receptor is mainly expressed in nociceptive sensory neurons in rat lumbar dorsal root ganglia

Several lines of evidence indicate that peripheral 5-HT2A receptors are involved in the development of inflammatory and neuropathic pain. However, their localization in sensory cell bodies is not accurately known. We therefore studied 5-HT2A receptor distribution in rat lumbar dorsal root ganglia using immunocytochemistry. Forty percent of L3 lumbar dorsal root ganglion cells were immunoreactive for 5-HT2A receptor. Most were small- to medium-sized cell bodies. Double-labeled experiments revealed that they expressed various chemical phenotypes. The smaller 5-HT2AR cell bodies often bind the isolectin B4 although some 5-HT2AR cell bodies also express substance P (SP). Many 5-HT2A-positive small dorsal root ganglion cells expressed the capsaicin receptor transient receptor potential vanilloid type 1 receptor (TRPV1), confirming their nociceptive nature. In addition, a few large cell bodies were labeled for 5-HT2A, and they also expressed NF200 suggesting that they were at the origin of Aδ or Aβ fibers. A total absence of double labeling with parvalbumin showed that they were not proprioceptors. 5-HT2A immunoreactivity in dorsal root ganglia cells was found in the cytoplasm and along the plasma membrane at the interface between sensory cell and the adjacent satellite cells; this distribution was confirmed under the electron microscope, and suggested a functional role for the 5-HT2A receptor at these sites. We therefore investigated the presence of 5-HT and 5-HIAA in lumbar dorsal root ganglia by high performance liquid chromatography. There were 5.75±0.80 ng 5-HT and 3.19±0.37 ng 5-hydroxyindoleacetic acid (5-HIAA) per mg of protein with a ratio 5-HIAA/5-HT of 0.67±0.10, similar to values typically observed in brain tissues. These findings suggest that 5-HT, via the 5-HT2AR, may be involved in the peripheral control of sensory afferents, mainly unmyelinated nociceptors and to a lesser extent neurons with Aδ or Aβ fibers, and in the control of cellular excitability of some dorsal root cell bodies through a paracrine mechanism of action.     

人胎小肠内胰岛淀粉样多肽及5-羟色胺免疫反应细胞的个体发生

目的 探讨胰岛淀粉样多肽免疫反应(IAPP-IR)细胞和5-羟色胺(5-HT)免疫反应细胞(EC细胞)在人胎小肠中的个体发生及IAPP与5-HT的关系。方法 免疫组织化学技术。结果 胎期小肠内IAPP-IR细胞仅见于十二指肠。16周开始，在十二指肠绒毛上皮细胞间可见单个散在的IAPP-IR细胞；22～27周，其细胞数量则逐渐增多，主要分布于肠腺中。EC细胞可见于胎期小肠各段，并随着胎龄增长，其细胞密度大小依次为十二指肠、空肠、回肠。11周，在小肠3段绒毛上皮和尚未分化完全的肠腺细胞间已可见该种细胞；17～21周，数量达最多，主要分布于绒毛根部和肠腺的上皮细胞问；22周后，EC细胞呈渐少趋势。免疫组织化学邻片单染比较，未见IAPP和5-HT在同一细胞内有共存现象。结论 IAPP、5-HT在人胎小肠的内分泌细胞中已有表达。IAPP-IR细胞及EC细胞随着胎儿的发育而发生不同变化。     

人胎阑尾IAPP-、SS-及5-HT免疫反应性细胞的免疫组织化学研究

为了探讨胰岛淀粉样多肽，在胎阑尾中的个体发及其与其他生物活性物质的关系，用免疫组织ＰＡＰ法，对３６例１２－３８周人胎阑尾中ＩＡＰＰ免疫反应性细胞，生称抑素ＩＲ细胞５－羟色胺－ＩＲ细胞进行定位研究。结果表明，肥１２周，阑尾上皮中已可见到ＳＳ－ＩＲ细胞，而ＩＡＰＰ－及５－ＨＴ－ＩＲ细胞于１４周才见到。在整个胎期，阑尾中的ＩＡＰＰ－及ＳＳ－ＩＲ细胞始络分散存在，数量较少，而５－ＨＴ－ＩＲ细胞数量随胎龄增     

大鼠神经系统内5-羟色胺_(1A)受体亚型的定位分布(英文)

应用免疫组织化学技术观察了大鼠神经系统内 5 -羟色胺 1A受体亚型 ( 5 -HT1 AR)免疫阳性结构的分布。结果显示 :5 -HT1 AR免疫阳性结构主要分布于梨状皮质、隔核、丘脑腹后核、丘脑网状核、杏仁基外侧核、Purkinje细胞层、红核、面神经核、斜方体核等 ;在海马、额叶皮质、丘脑背内侧核、脚间核、三叉神经中脑核、中缝背核、三叉神经脊束核、脊髓背角浅层、背根神经节和三叉神经等结构内有中等强度的分布 ;在嗅球、尾壳核、苍白球、斜角带核、终纹床核、缰核、黑质、上橄榄等部位有弱的分布。本文的结果提示 5 -HT1 AR阳性结构广泛地分布在大鼠神经系统 ,它们可能介导 5 -HT在神经系统中的多种生理功能     

人胎儿结肠及直肠内胰岛淀粉样多肽及5-羟色胺免疫反应细胞的研究

目的 探讨胰岛淀粉样多肽(IAPP)在人胎结肠、直肠中的个体发生及其与其他生物活性物质的关系。方法 用免疫组织化学SABC法，对31例9～27周人胎结肠及直肠中IAPP免疫反应(IR)细胞和5-羟色胺(5-HT)-IR细胞进行定位研究。结果 人胎9周结肠内已可见较多的5-HT-IR细胞，而IAPP-IR细胞于18周出现，直肠内5-HT-IR细胞和IAPP-IR细胞均于11周出现。随胎龄增长，5-HT-IR细胞由少至多，20周达到高峰，21周后逐渐减少；而IAPP-IR细胞在整个胎期始终分散存在，数量较少。经邻片比较观察发现，IAPP-IR细胞与部分5-HT-IR细胞定位一致。免疫组织化学双染法显示有的细胞内IAPP与5-HT共存。结论 胎儿期结肠及直肠的内分泌细胞已开始合成IAPP，并在部分细胞内IAPP和5-HT有共存。     

Immunolocalization of islet amyloid polypeptide (IAPP) in pancreatic beta cells by means of peroxidase-antiperoxidase (PAP) and protein A-gold techniques.

A novel putative polypeptide hormone identified as islet amyloid polypeptide (IAPP) was recently purified from islet amyloid (IA) of diabetic humans and cats, and also from amyloid of a human insulinoma. Although the function of IAPP is yet unknown, its occurrence in pancreatic endocrine tissue and its partial amino acid sequence identity with calcitonin gene-related peptide (CGRP) suggests an endocrine regulatory effect. In the present investigation, the authors utilized antisera to insulin, glucagon, somatostatin, pancreatic polypeptide, synthetic human CGRP, and a synthetic human IAPP (7-17) undecapeptide to immunohistochemically (PAP technique) document the presence of IAPP immunoreactive cells in the islets of the cat, dog, mouse, and rat, but not in the islets of the horse or calf. In serial sections of islets from these species it was shown that IAPP immunoreactivity occurred in insulin-reactive beta cells. This observation was confirmed immunocytochemically in cat islets by means of protein A-gold probes. With protein A-gold labeling techniques, IAPP immunoreactivity was localized to the outer lucent compartment of the beta cell secretory granule, whereas insulin immunoreactivity was associated with the electron-dense core. These findings provide strong evidence that IAPP or an IAPP precursor is synthesized by beta cells and is stored in beta cell granules for subsequent co-secretion with insulin. The conservation of IAPP in humans and multiple animal species and the localization of IAPP to pancreatic beta cells provide further evidence that IAPP has an important endocrine regulatory function. The propensity of IAPP to polymerize and form IA fibrils in diabetes associated with aging may indicate that IAPP is in some way also linked to the development of Type 2 diabetes.     

大鼠耳郭神经肽Y,降钙素基因相关肽和5—羟色胺免疫反应性神经起源

以CB-HRP逆行追踪与免疫细胞化学法相结合,研究了大鼠耳郭局部神经肽Y、降钙素基因相关肽和5-羟色胺免疫反应性神经的来源;结果:在面神经核内和C_(2～3)脊神经节仅见降钙素基因相关肽-CBHRP一种双标细胞,神经肽Y和5-羟色胺免疫反应在面神经和C_(2～3)脊神经节则呈阴性.在颈上神经节神经肽Y-CBHRP、降钙素基因相关肽-CBHRP和5-羟色胺-CBHRP三种标细胞均有出现.文内讨论了耳郭微循环常用观察部位的运动、感觉以及微循环调节的神经生理机制.     

Human islet amyloid polypeptide expression in COS-1 cells. A model of intracellular amyloidogenesis.

Non-insulin-dependent diabetes mellitus is characterized by concurrent loss of beta-cells and deposition of islet amyloid derived from islet amyloid polypeptide (IAPP). We have previously demonstrated that IAPP-derived amyloid forms intracellularly in humans with chronic excess insulin expression (eg, insulinoma and insulin receptor antibody-induced insulin resistance). To determine whether overexpression of IAPP results in intracellular amyloid in mammalian cells, we transfected COS cells with vectors expressing amyloidogenic human IAPP or non-amyloidogenic rat IAPP. Transfected COS-1 cells secreted comparable amounts of human IAPP and rat IAPP (2.1 to 2.8 nmol/L/48 hours). After 96 hours, 90% of cells expressing human IAPP contained amyloid fibrils and were degenerating or dead, whereas cells transfected with rat IAPP lacked amyloid and were viable. Thus, overexpression of human IAPP can result in intracellular amyloid formation that is associated with cell death, suggesting that intracellular amyloid may play a role in beta-cell loss in non-insulin-dependent diabetes mellitus.