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Immunohistochemical analysis for neural markers of the lateral retinaculum in patients with isolated symptomatic patellofemoral malalignment -: A neuroanatomic basis for anterior knee pain in the active young patient

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AMERICAN JOURNAL OF SPORTS MEDICINE 2000年第5期28卷 725-731页

作者： Sanchis-Alfonso, V Roselló-Sastre, E Hosp Arnau Vilanova Dept Orthopaed Surg Valencia Spain Hosp Univ Dr Peset Dept Pathol Valencia Spain

We evaluated 13 lateral retinacula excised at the time of Insall proximal realignments or isolated lateral retinacular releases performed in patients with isolated symptomatic patellofemoral malalignment recalcitrant to nonoperative treatment. Evaluation was performed by means of conventional histologic and immunohistochemical analysis for neural markers (S-100 protein, neurofilament protein, substance P, and neural growth factor). The observations reported here provide a neuroanatomic basis for anterior knee pain syndrome in active young patients with isolated symptomatic patellofemoral malalignment and support the clinical observation that the lateral retinaculum may have a key role in the origin of this pain as a result of increased neural growth factor production, which induces proliferation of nociceptive axons, mainly in a perivascular location.

关键词：生物学标记/分析免疫组织化学膝关节/解剖学和组织学膝关节/神经支配膝关节/病理学神经组织蛋白质类/分析疼痛/病理生理学髌骨/畸形综合征青少年成年人女(雌)性人类男(雄)性

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Molecular dissection of regulated secretory pathways in human gastric enterochromaffin-like cells:: an immunohistochemical analysis

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HISTOCHEMISTRY AND CELL BIOLOGY 1999年第3期112卷 205-214页

作者： Höcker, M John, M Anagnostopoulos, J Buhr, HJ Solimena, M Gasnier, B Henry, JP Wang, TC Wiedenmann, B Humboldt Univ Klinikum Charite Med Klin Schwerpunkt Hepatol & Gastroenterol D-13353 Berlin Germany Free Univ Berlin Klinikum Benjamin Franklin Med Klin 1 D-12002 Berlin Germany Free Univ Berlin Klinikum Benjamin Franklin Abt Pathol D-12002 Berlin Germany Free Univ Berlin Klinikum Benjamin Franklin Allgemeinchirurg Abt D-12002 Berlin Germany Yale Univ Sch Med Div Med New Haven CT USA Yale Univ Sch Med Div Endocrinol New Haven CT USA Inst Biol Phys Chim F-75005 Paris France Harvard Univ Massachusetts Gen Hosp Sch Med Gastroenterol Unit Boston MA USA Harvard Univ Massachusetts Gen Hosp Sch Med Dept Internal Med Boston MA USA

Enterochromaffin-like (ECL) cells regulate gastric acid secretion through vesicular release of histamine. Until now, the molecular machinery of human ECL cells involved in the formation and release of vesicles is largely unknown. We analyzed tissue samples obtained from normal human gastric mucosa (n=4) and ECLomas (n=5) immunohistochemically using the APAAP method or double immunofluorescence confocal laser microscopy. Human pheochromocytomas (n=5) were investigated in parallel and compared to ECL cells. Secretory pathways were characterized using antibodies specific for marker proteins of large dense-core vesicles (LDCVs;islet cell antigen 512, chromogranin A, pancreastatin, and vesicular monoamine transporter 2) and small synaptic vesicle (SSV) analogues (synaptophysin). Tissues were also analyzed for expression of the peptide hormone processing enzymes, carboxypeptidase E and prohormone convertase 1, as well as the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, 25-kDa synaptosome-associated protein (SNAP25), syntaxin, and syn aptobrevin. Immunoreactivity for markers of LDCVs and SSV analogues were detected in normal ECL cells and ECLomas. Both tissues also showed expression of carboxypeptidase E and prohormone convertase 1. Analysis of vesicular SNARE (v-SNARE) and target membrane SNARE (t-SNARE) proteins revealed the presence of SNAP25, syntaxin, and synaptobrevin in normal and neoplastic ECL cells. Our data suggest that ECL cells possess the two vesicle types of regulated neuroendocrine secretory pathways, LDCVs and SSV analogues. Since ECL cells also contain typical SNARE proteins, the molecular machinery underlying secretory processes in this cell type appears to be identical to the secretory apparatus of neuroendocrine cells and neurons. In addition, our findings suggest that the secretory apparatus of ECL cells is maintained during neoplastic transformation.

关键词：天冬氨酸内肽酶类/分析生物学标记/分析羧肽酶H 羧肽酶类/分析载体蛋白质类/分析细胞质颗粒/化学细胞质颗粒/酶学肠嗜铬样细胞/化学肠嗜铬样细胞/酶学肠嗜铬样细胞/病理学肠嗜铬样细胞/分泌荧光抗体技术间接胃黏膜/化学胃黏膜/酶学胃黏膜/病理学胃黏膜/分泌免疫酶技术膜蛋白质类/分析神经组织蛋白质类/分析嗜铬细胞瘤/化学嗜铬细胞瘤/酶学嗜铬细胞瘤/病理学嗜铬细胞瘤/分泌前蛋白转化酶类 Qa-SNARE蛋白质类 Qc-SNARE蛋白质类 R-SNARE蛋白质类胃肿瘤/化学胃肿瘤/酶学胃肿瘤/病理学胃肿瘤/分泌突触体相关蛋白质25 膜泡运输蛋白质类人类

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Radiation-induced apoptosis in the neonatal and adult rat spinal cord

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RADIATION RESEARCH 2000年第3期154卷 268-276页

作者： Li, YQ Wong, CS Univ Toronto Univ Hlth Network Princess Margaret Hosp Dept Radiat Oncol Toronto ON M5G 2M9 Canada Univ Toronto Div Expt Therapeut Toronto ON M5G 2M9 Canada

This study was designed to characterize radiation-induced apoptosis in the spinal cord of the neonatal and young adult rat. Spinal cords (C2-T2) of 1-, 2 and 10-week-old rats were irradiated with a single dose of 8, 18 or 22 Gy, Apoptosis was assessed histologically according to its specific morphological features or by using the TUNEL assay. Cell proliferation was assessed immunohistochemically using BrdU, Identities of cell types undergoing apoptosis were assessed using immunohistochemistry or in situ hybridization using markers for neurons, glial progenitor cells, microglia, oligodendrocytes and astrocytes, The time course of radiation-induced apoptosis in 1- or 2-week-old rat spinal cord was similar to that in the young adult rat spinal cord. A peak response was observed at about 8 h after irradiation, and the apoptosis index returned to the levels in nonirradiated spinal cords at 24 h, The neonatal rat spinal cord demonstrated increased apoptosis compared to the adult, Values for total yield of apoptosis over 24 h induced by 8 Gy in the neonatal rat spinal cord were significantly greater than that in the adult. Immunohistochemistry studies using Leu7, galactocerebroside, Rip and adenomatous polyposis coli tumor suppressor protein indicated that most apoptotic cells were cells of the oligodendroglial lineage regardless of the age of the animal. No evidence of Gfap or factor VIII-related antigen positive apoptotic cells was observed, and there was a small number of apoptotic microglial cells (lectin-Rca1 positive) in the neonatal and adult rat spinal cord. In the neonatal but not adult rat spinal cord, about 10% of the apoptotic cells appeared to be neurons and were immunoreactive for synaptophysin, Labeling indices (LI) for BrdU in nonirradiated 1- and 2-week-old rat spinal cord were 20.0 and 16.3%, respectively, significantly greater than the LI of 1.0% in the 10-week-old rat spinal cord. At 8 h after a single dose of 8 Gy, 13.4% of the apoptotic cells were Brd

关键词：年龄因素动物新生细胞凋亡/辐射效应细胞分裂/辐射效应因子Ⅷ/分析神经胶质原纤维酸性蛋白质/分析原位杂交原位缺口末端标记神经组织蛋白质类/分析神经胶质/病理学神经胶质/辐射效应神经元/病理学神经元/辐射效应辐射损伤实验性/病理学辐射耐受性大鼠近交F344 脊髓/生长和发育脊髓/病理学动物女(雌)性大鼠

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A hippocampal interneuron associated with the mossy fiber system

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第3期97卷 1275-1280页

作者： Vida, I Frotscher, M Univ Freiburg Inst Anat D-79104 Freiburg Germany

Network properties of the hippocampus emerge from the interaction of principal cells and a heterogeneous population of interneurons expressing gamma-aminobutyric acid (GABA). To understand these interactions, the synaptic connections of different types of interneurons need to be elucidated. Here we describe a type of inhibitory interneuron of the hippocampal CA3 region that has an axon coaligned with the mossy fibers. Whole-cell patch-clamp recordings, in combination with intracellular biocytin filling, were made from nonpyramidal cells of the stratum lucidum under visual control. Mossy fiber-associated (MFA) interneurons generated brief action potentials followed by a prominent after-hyperpolarization. Subsequent visualization revealed an extensive axonal arbor which was preferentially located in the stratum lucidum of CA3 and often invaded the hilus, The dendrites of MFA interneurons were mainly located in the strata radiatum and oriens, suggesting that these cells are primarily activated by associational and commissural fibers. Electron microscopic analysis showed that axon terminals of MFA interneurons established symmetric synaptic contacts predominantly on proximal apical dendritic shafts. and to a lesser degree, on somata of pyramidal cells. Synaptic contacts were also found on GABAergic interneurons of the CA3 region and putative mossy cells of the hilus, Inhibitory postsynaptic currents (IPSCs) elicited by MFA interneurons in simultaneously recorded pyramidal cells had fast kinetics (half duration, 3.6 ms) and were blocked by the GABA(A) receptor antagonist bicuculline, Thus, MFA interneurons are GABAergic cells in a position to selectively suppress the mossy fiber input, an important requirement for the recall of memory traces from the CA3 network.

关键词：动作电位传入通路/超微结构轴突/超微结构树突/超微结构海马/细胞学中间神经元/化学中间神经元/细胞学中间神经元/生理学显微镜检查电视苔藓纤维海马/超微结构神经组织蛋白质类/分析膜片钳术大鼠 Wistar 受体 GABA-B/分析 γ氨基丁酸/分析动物大鼠

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Neurodegeneration with brain iron accumulation, type 1 is characterized by α-, β-, and γ-synuclein neuropathology

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AMERICAN JOURNAL OF PATHOLOGY 2000年第2期157卷 361-368页

作者： Galvin, JE Giasson, B Hurtig, HI Lee, VMY Trojanowski, JQ Univ Penn Sch Med Ctr Neurodegenerat Dis Res Dept Pathol & Lab Med Philadelphia PA 19104 USA Univ Penn Dept Neurol Philadelphia PA 19104 USA Med Coll Penn & Hahnemann Univ Dept Neurol Philadelphia PA 19102 USA

Neurodegeneration with brain iron accumulation, type 1 (NBIA 1), or Hallervorden-Spatz syndrome, is a rare neurodegenerative disorder characterized clinically by Parkinsonism, cognitive impairment, pseudobulbar features, as well as cerebellar ataxia, and neuropathologically by neuronal loss, gliosis, and iron deposition in the globus pallidus, red nucleus, and substantia nigra. The hallmark pathological lesions of NBIA 1 are axonal spheroids, but Lewy body (LB)-like intraneuronal inclusions, glial inclusions, and rare neurofibrillary tangles also occur. Here we show that there is an accumulation of alpha-synuclein (alpha S) in LB-like inclusions, glial inclusions, and spheroids in the brains of three NBIA 1 patients. Further, beta-synuclein (beta S) and gamma-synuclein (gamma S) immunoreactivity was detected in spheroids but not in LB-like or glial inclusions. Western blot analysis demonstrated high-molecular weight alpha S aggregates in the high-salt-soluble and Triton X-100-insoluble/sodium dodecyl sulfate-soluble fraction of the NBIA 1 brain. Significantly, the levels of alpha S were markedly reduced in the Triton X-100-soluble fractions compared to control brain, and unlike other synucleinopathies, insoluble alpha S did not accumulate in the formic acid-soluble fraction. These findings expand the concept of neurodegenerative synucleinopathies by Implicating alpha S, beta S, and gamma S in the pathogenesis of NBIA 1.

关键词：淀粉样β肽类/分析印迹法蛋白质脑/代谢脑/病理学致命性结局免疫组织化学铁/代谢神经组织蛋白质类/分析神经变性疾病/代谢神经变性疾病/病理学突触核蛋白类 α突触核蛋白 β突触核蛋白 γ突触核蛋白 tau蛋白质类/分析青少年老年人人类婴儿

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POPULATION PARAMETERS - ACQUISITION AND ANALYSIS OF ELECTROPHORETIC BRAIN PROTEIN PATTERNS BY MINICOMPUTER-COUPLED DENSITOMETRY

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ANALYTICAL BIOCHEMISTRY 1975年第2期69卷 350-360页

作者： MAYOR, SJ MED COLL OHIO DEPT PHYSIOLTOLEDOOH 43614

Interfacing a minicomputer with a high resolution densitometer permits the online acquisition and statistical analysis of population parameters for experimental and control sample patterns of various macromolecules obtained by polyacrylamide gel electrophoresis. This technique, illustrated in this paper using protein from the brains of Japanese quail, enables the experimenter to make comparisons which were previously not readily available for electrophoresis.

关键词：脑化学计算机光密度测定法/方法电泳聚丙烯酰氨凝胶/方法神经组织蛋白质类/分析鹌鹑动物男(雄)性

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Evidence that endogenous thymosin alpha-1 is present in the rat central nervous system

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NEUROCHEMISTRY INTERNATIONAL 1999年第6期35卷 463-470页

作者： Turrini, P Aloe, L CNR Inst Neurobiol I-00137 Rome Italy

We have previously reported that administration of Thymosin alpha 1 (T-alpha 1) can enhance the level of the Nerve Growth Factor and the distribution of its receptor in the developing Central Nervous System (CNS) of rat. To further explore the role of T-alpha 1 and verify its presence in cells of rat CNS, we carried out an immunohistochemical study using a polyclonal antibody against T-alpha 1. T-alpha 1 immunoreactivity was found mainly in neurons of the hippocampus and spinal cord and in several small cells, resembling glial cells, of specific regions of the brain. Moreover, to study whether cerebral cells were receptive to T-alpha 1, we injected iodinated T-alpha 1 (I-125-T-alpha 1) icv. I-125-T-alpha 1 labelled neurons were observed in the hypothalamus and septal nuclei. Our results indicate that specific neuronal populations in the rat CNS are able to express and respond to T-alpha 1. (C) 1999 Elsevier Science Ltd. All rights reserved.

关键词：脑化学齿状回/化学荧光抗体技术间接海马/化学免疫血清免疫酶技术运动神经元/化学神经组织蛋白质类/分析神经胶质/化学神经免疫调节神经元/化学锥体细胞/化学大鼠 Sprague-Dawley 隔核/化学脊髓/化学胸腺素/类似物和衍生物胸腺素/分析胸腺素/免疫学胸腺素/药代动力学动物女(雌)性男(雄)性兔大鼠

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Marked impairment of protein tyrosine phosphatase 1B activity in adipose tissue of obese subjects with and without type 2 diabetes mellitus

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JOURNAL OF LABORATORY AND CLINICAL MEDICINE 1999年第2期134卷 115-123页

作者： Cheung, A Kusari, J Jansen, D Bandyopadhyay, D Kusari, A Bryer-Ash, M Univ Tennessee Coll Med Dept Med Memphis TN USA Vet Adm Med Ctr Res Serv Memphis TN 38104 USA Tulane Univ Sch Med Dept Physiol New Orleans LA 70112 USA Tulane Univ Sch Med Dept Surg New Orleans LA 70112 USA

Protein tyrosine phosphatases (PTPs) are required for the dephosphorylation of the insulin receptor (IR) and its initial cellular substrates, and it has recently been reported that PTP-1B may play a role in the pathogenesis of insulin resistance in obesity and type 2 diabetes mellitus (DM), We therefore determined the amount and activity of PTP-1B in abdominal adipose tissue obtained from lean nondiabetic subjects (lean control (LC)), obese nondiabetic subjects (obese control (OC)), and subjects with both type 2 DM (DM2) and obesity (obese diabetic (OD)), PTP-1B protein levels were 3-fold higher in OC than in LC (1444 +/- 195 U vs 500 +/- 146 U (mean +/- SEM), P < .015), while OD exhibited a 5.5-fold increase (2728 +/- 286 U, P < .01), PTP activity was assayed by measuring the dephosphorylating activity toward a phosphorus 32-labeled synthetic dodecapeptide, In contrast to the increased PTP-1B protein levels, PTP-1B activity per unit of PTP-1B protein was markedly reduced, by 71% and 88% in OC and On, respectively. Non-PTP-1B tyrosine phosphatase activity was comparable in all three groups. Similar results were obtained when PTP-1B activity was measured against intact human IR, A significant correlation was found between body mass index (BMI) and PTP-1B level (r = 0.672, P < .02), whereas BMI and PTP-1B activity per unit of PTP-1B showed a strong inverse correlation (r = -0.801, P < .002). These data suggest that the insulin resistance of obesity and DM2 is characterized by the increased expression of a catalytically impaired PTP-1B in adipose tissue and that impaired PTP-1B activity may be pathogenic for insulin resistance in these conditions.

关键词：脂肪组织/酶学印迹法蛋白质细胞系糖尿病 2型/酶学酶激活/生理学成纤维细胞/细胞学水解作用神经组织蛋白质类/分析神经组织蛋白质类/代谢肥胖症/酶学磷放射性同位素/诊断应用磷酰化沉淀素试验成年人老年人动物女(雌)性人类男(雄)性中年人大鼠

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Vesicle-associated proteins and P2X receptor clusters at single sympathetic varicosities in mouse vas deferens

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JOURNAL OF NEUROCYTOLOGY 1999年第6期28卷 469-480页

作者： Barden, JA Cottee, LJ Bennett, MR Univ Sydney Inst Biomed Res Sydney NSW 2006 Australia Univ Sydney Dept Anat & Histol Sydney NSW 2006 Australia Univ Sydney Dept Physiol Sydney NSW 2006 Australia

1. Antibodies against vesicle-associated proteins of the SNARE complex (syntaxin (AbS), SNAP 25 (AbS25), synaptobrevin (VAMP;AbV) and the alpha 1B subunit of calcium channels (Ab alpha(1B)) were located with respect to sympathetic varicosities (labelled with the ubiquitous vesicle proteoglycan antibody AbSV2) and to clusters of P2X receptor subunits (labelled with antibodies AbP2X(1) to AbP2X(6)). In addition, these receptor clusters were located with respect to Schwann cells labelled with antibodies to S100 (AbS100). 2. The spatial relation between proteins of the SNARE complex and calcium channels was determined. AbS25 patches ranged from 250-500 nm in size and were often colocalised with smaller AbS patches (250-350 nm). Ab alpha(1B) patches (300-700 nm diameter) were always coincidental with AbS patches. AbV patches (400-1000 nm in diameter) also coincided with AbS patches. 3. The spatial relation between different P2X subunit clusters and varicosities labelled with AbSV2 was ascertained. Large (500-700 nm diameter) AbP2X(1) receptor clusters were found colocalised with many (91%) AbSV2 labelled varicosities, although small diameter (250-350 nm) AbP2X(1) clusters occurred at random over the muscle. Small AbP2X(2) clusters were found uniquely in the vicinity of AbSV2 labelled varicosities, but were not entirely coincidental with these. Small AbP2X(3) receptor clusters were found in close association with AbSV2 labelled nerves. Small diameter AbP2X(4) clusters (250-350 nm) were found throughout the muscle with some of these coincidental with AbSV2 labelling. Small diameter AbP2X(5) (250-350 nm) cluster labelling was found in juxtaposition to strings of AbSV2 labelled varicosities but were not coincidental with these. Small (250-350 nm) diameter AbP2X(6) clusters were also found in close juxtaposition to AbSV2 labelled nerves. 4. The spatial relation between different P2X subtype clusters and Schwann cells labelled with AbS100 was examined. Both AbP2X(1) and AbP2X(

关键词：钙通道/分析膜蛋白质类/分析显微镜检查共焦神经组织蛋白质类/分析神经元/细胞学卵母细胞/细胞学 Qa-SNARE蛋白质类受体嘌呤能P2/分析重组蛋白质类/分析 SNARE蛋白质类许旺细胞/细胞学交感神经系统/细胞学突触体相关蛋白质25 转染输精管/神经支配膜泡运输蛋白质类爪蟾光滑动物男(雄)性小鼠

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A comparison of synaptic protein localization in hippocampal mossy fiber terminals and neurosecretory endings of the neurohypophysis using the cryo-immunogold technique

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JOURNAL OF NEUROCYTOLOGY 2000年第1期29卷 19-30页

作者： Zhang, LX Volknandt, W Gundelfinger, ED Zimmermann, H Goethe Univ Frankfurt Biozentrum AK Neurochem D-60439 Frankfurt Germany Leibniz Inst Neurobiol D-39118 Magdeburg Germany

In central synapses synaptic vesicle docking and exocytosis occurs at morphologically specialized sites (active zones) and requires the interaction of specific proteins in the formation of a SNARE complex. In contrast, neurosecretory terminals lack active zones. Using the cryo-immunogold technique we analyzed the localization of synaptic vesicle proteins and of proteins of the docking complex at active zones. This was compared to the localization of the identical proteins in neurosecretory terminals. In addition we compared the vesicular and granular localization of the proteins investigated. Synaptic vesicles in rat hippocampal mossy fiber synapses and microvesicles in the neurosecretory terminals of the neurohypophysis contained in common the proteins VAMP II (a v-SNARE), SV2, rab3A, and N-type Ca2+ channels. Only minor immunolabeling for these proteins was observed at neurosecretory granules. These results support the notion of a close functional identity of microvesicles from neurosecretory endings of the neurohypophysis and of synaptic vesicles. The vesicular pool of N-type Ca2+ channels may serve their stimulation-induced translocation into the plasma membrane. We find increased labeling for VAMP II, SNAP-25, N-type Ca2+ channels and of rab3A at the active zones of mossy fiber synapses. Labeling at release sites is by far highest for Bassoon, a high molecular weight protein of the active zone. The labeling pattern implies an association of Bassoon with presynaptic dense projections. Bassoon is absent from neurosecretory terminals and VAMP II, SNAP-25, rab3A, and N-type Ca2+ channels reveal a scattered distribution over the plasma membrane. The competence of the presynaptic active zone for selective vesicle docking may not primarily result from its contents in SNARE proteins but rather from the preformation of presynaptic dense projections as structural guides for vesicle exocytosis.

关键词：钙通道 N型/分析免疫组织化学/方法膜蛋白质类/分析显微镜检查免疫电子/方法苔藓纤维海马/化学苔藓纤维海马/超微结构神经组织蛋白质类/分析垂体后叶/化学垂体后叶/超微结构 R-SNARE蛋白质类大鼠 Wistar SNARE蛋白质类突触小泡/化学突触小泡/超微结构突触体相关蛋白质25 膜泡运输蛋白质类 rab3AGTP结合蛋白质/分析动物大鼠

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