Immunohistochemical Localization of Calmodulin-Dependent Cyclic Phosphodiesterase in the Human Brain

The amplification of cyclic nucleotide second messenger signals within neurons is controlled by phosphodiesterases which are responsible for their degradation. Calmodulin-dependent phosphodiesterase (CaMPDE) is an abundant enzyme in brain which carries out this function. For the first time, we have localized CaMPDE in the normal human brain at various ages, using a monoclonal antibody designated A6. This antibody was generated using standard techniques, purified, and applied to tissue sections. Autopsy specimens of human brain with no neuropathological abnormalities were selected representing a range of pre- and postnatal ages. Sections of various brain regions were evaluated for immunoreactivity, graded as nil, equivocal, or definite. We demonstrated definite CaMPDE immunohistochemical staining in neocortex, especially in neurons in layers 2 and 5. There was definite neuronal immunoreactivity in the hippocampus, and in the subiculum. The striatum had definite patchy neuronal staining. Definite terminal staining in the globus pallidus externa and substantia nigra pars reticulata outlined resident neurons, interpreted as axonal terminal staining. Cerebellar Purkinje cells showed definite immunoreactivity. In the developing brain, definite immunohistochemical staining was seen in the cerebellar external granular layer. The expression of CaMPDE in specific subsets of neurons suggests they may correlate with cells having dopaminergic innervation and/or high levels of neuronal integration.     

钙调素激酶在玉米体内的免疫组织化学定位

应用免疫组织化学定位方法研究了玉米体内钙调素激酶(CaM kinase,CaMK)的表达模式。结果表明CaMK广泛分布于玉米体内,但表达水平存在着明显时空差异。在营养器官中钙调素激酶主要分布于叶的维管束鞘细胞、侧根原基和根尖等部位,而其它部位没有检测到明显的分布。在生殖器官中,有大量钙调素激酶分布于幼胚及花药小孢子母细胞、四分体及绒毡层细胞中;在成熟胚囊的卵细胞、中央细胞以及二者的分界面上也有少量分布。这些结果为进一步探索钙调素激酶在植物体内的生理功能提供了重要线索。     

Lymphatic and Blood Vessels of the Aged Human Gingiva1

The lymphatic pathways in the gingivae from aged humans were traced by the use of the PAS reaction or iron hematoxylin stain, and their structural characteristics were compared to those of the blood microvasculature. In the young and aging gingivae the lymphatic capillaries originated in the connective tissue papillae of the lamina propria, and appeared as thin walled irregular shaped vessels. The adjacent blood capillaries in aged gingivae differed in that their walls were thicker and stained intensely Schiff positive than seen in young adult gingivae. The lymphatic capillaries emptied into thin walled collecting vessels of varying calibers that course through the lamina propria to reach the main conducting vessels that contained valves projecting within its lumen. The accompanying blood vessels were easily differentiated from the lymphatic vessels by the intense positive staining of their walls following exposure to the PAS reaction. Distended lymphatic vessels of different caliber were demonstrable in inflamed aging gingivae, suggesting that lymphatic vessels in the aged gingivae were able to provide a drainage system for excessive fluid, proteins, and other particulates from both non-injured and injured sites.     

血管紧张素II在小鼠卵母细胞中的免疫组化定位(英文)

本文研究了血管紧张素II在小鼠卵母细胞中的免疫组织化学定位。结果表明血管紧张素II不仅分布在卵巢内的黄体细胞、卵泡的膜细胞、基质和血管,在卵母细胞的细胞质和细胞膜上也见有阳性分布。颗粒细胞和卵丘细胞上未见着色。在恢复减数分裂过程中,处于生发泡破裂和第一极体排放期的卵母细胞内也检测到血管紧张素II[(\265\304\303\342\322\337\321\364\320\324\316\357\241\243)238.1(\322\362\264\313)],血管紧张素II有可能在卵泡的生长发育和卵母细胞的成熟过程中起着重要作用。     

小鼠卵巢促性腺激素受体的免疫组化定位

目的探明小鼠两种促性腺激素受体(FSHr、LHr)在卵巢的位置分布,揭示促性腺激素(GTH)调节卵巢机制及与卵泡发育分化的关系。方法运用免疫组化ABC法对小鼠卵巢FSHr、LHr分别进行定位染色,结合图像分析系统处理分析阳性切片。结果①FSHr阳性物质主要见于GC、TC、卵母细胞及间质细胞。随卵泡的发育,FSHr、LHr阳性细胞数量呈增长趋势,卵泡早期与中期之间阳性细胞数量差异显著,平均吸光度变化差异不显著。②LHr阳性物质主要见于卵泡TC、间质细胞、GC、卵母细胞,阳性物质着色以卵泡中、晚期较强,阳性细胞数量以卵泡中期与晚期之间差异显著。平均吸光度变化差异不显著。结论卵泡颗粒细胞、膜细胞上受体是接受促性腺激素的主要调节部位,受体数量与卵泡大小和发育程度有一定的正相关。     

Immunohistochemical Localization of Cyclic GMP in Aggregating Polysphondylium violaceum

Using indirect immunofluorescence technique, it has been possible to localize cyclic GMP in Polysphondylium violaceum cells. The bound cyclic nucleotide is localized throughout the cell during early stages, however, this staining increases and there is marked localization of cyclic GMP in the nuclear areas of the cells when aggregation is in full swing. Over 90% of the cells exhibited intense nuclear staining by 6 h and this decreased to less than 10% by 10 h.     

青紫蓝兔体内Ghrelin的免疫组化定位

采用免疫组织化学方法研究了Ghrelin在青紫蓝兔(Oryctolagus cuniculus)体内的分布定位。结果显示,Ghrelin免疫阳性细胞分布于下丘脑、大脑皮质、延髓、脊髓、胃、小肠和大肠。下丘脑内的阳性神经元胞体主要分布于弓状核、室旁核、腹内侧核、背内侧核和下丘脑外侧区。胃肠道内的Ghrelin免疫阳性细胞存在两种类型,即"闭合型"细胞和"开放型"细胞。实验结果表明,兔体内Ghrelin的分布与人和其他动物的分布基本相似,但也存在一些差异。     

Immunohistochemical Localization of Bone Morphogenetic Proteins (BMPs) and their Receptors in Solitary and Multiple Human Osteochondromas

The expression of bone morphogenetic proteins (BMPs) and their cognate receptors (BMPRs) in osteochondromas has not been investigated. We determined the immunohistochemical localization and distribution of BMP-2/4, -6 and -7; BMP receptors BMPR-1A, BMPR-1B and BMPR-2; signal transducing proteins phosphorylated Smad1/5/8; and BMP antagonist noggin in the cartilaginous cap of solitary (SO) and multiple (MO) human osteochondromas and compared these with bovine growth plate and articular cartilage. The distribution and localization patterns for BMP-6, BMP-7, BMPR-1A and BMPR-2 were similar between the cartilaginous cap and the growth plate. BMP-2/4 and BMPR-1B were present throughout the growth plate. However, BMP-2/4 and phosphorylated Smad1/5/8 were mainly detected in proliferating chondrocytes of the cartilaginous cap. Also, BMPR-1B was found in hypertrophic chondrocytes of SO and proliferating chondrocytes of MO. Noggin was observed in resting chondrocytes and, to a lesser extent, in clustered proliferating chondrocytes in SO. On the other hand, noggin in MO was observed in proliferating chondrocytes. Since BMPs can stimulate proliferation and hypertrophic differentiation of chondrocytes, these findings suggest that there is an imbalance of BMP-2/4 and noggin interactions that may lead to abnormal regulation of chondrocyte proliferation and differentiation in the cartilaginous cap of human osteochondromas.     

Immunohistochemical Localization of Nerve Fibers in the Pseudocapsule of Fibroids

The pseudocapsule surrounding fibroids consists of compressed myometrium containing nerves and blood vessels that continue into adjacent myometrium. Oxytocin (OXT) is thought to affect wound healing after myomectomy. We determined the presence of OXT and protein gene product 9.5 (PGP9.5) immunoreactive nerve fibers in pseudocapsule compared to adjacent myometrium. Samples (N=106) of pseudocapsule and adjacent myometrium were collected from 57 women with uterine fibroids undergoing myomectomy, and stained with anti-OXT and PGP 9.5 antibodies to demonstrate the presence of nerve fibers. Nerve fibers in the pseudocapsule stained positively with OXT (89/106, 84.0%) and PGP 9.5 (94/106, 88.7%). The densities of nerve fibers staining with PGP 9.5 and OXT in the pseudocapsule were highest in the isthmus (23.68±22.45/mm² and 43.35±40.74/mm², respectively). There were no significant differences in the density of nerve fibers, stained with either OXT or PGP 9.5, between the pseudocapsule, and adjacent normal myometrium regardless of the fibroid location in the uterus (P>0.05). These results suggest that the pseudocapsule should avoid to be damaged during the myomectomy procedure.Key words: fibroid pseudocapsule, nerve fibers, oxytocin, myomectomy, protein gene product 9.5, immunohistochemistry     

Immunohistochemical Localization of Neuropeptides and Neurotransmitters in the Nucleus Solitarius

The nucleus tractus solitarii (NTS), which receives visceralafferent information from the cardiovascular, respiratory, gastrointestinaland taste systems, contains multiple neurotrasmitters and neuropeptidesthroughout its rostral to caudal extent. The neurotransmittersand neuropeptides immunoreactivity is located predominatelyin varicose fibers and small puncta throughout the neuropil.In addition, immunoreactive NTS neurons for a variety of neurotransmittersand neuropeptides are present in subnuclear regions. The neuroactive substances localized immunohistochemically inthe NTS include acetylcholine, the neuropeptides, substanceP, methionine- and leucine-enkephalin, ß-endorphin,cholecystokinin, neurotensin, galanin, calcitonin gene-relatedpeptide, somatostatin, FMRMamide, neuropeptide Y, angiotensinII, vasoactive intestinal polypeptide, vasopressin, oxytocin,thyrotropin-releasing hormone, luteinizing hormone-releasinghormone, atrial natriuretic peptide, the catecholamines, dopamine,norepinephrine, epinephrine, serotonin, histamine and the aminoacids, GABA and glutamate. The pattern of innervation for eachneurotransmitter and neuropeptide is not homogeneously distributedthroughout the NTS. Each substance has a unique pattern withinthe NTS as each subnuclear region contains different immunohistochemicalstaining patterns and densities of fibers. At the ultrastructural level both neurotransmitters and neuropeptidesare present in synaptic terminals that are in contact with differentparts of the neuronal membranes. Typically, the labeled terminalscontain both small, clear vesicles and large, dense core vesicleswith the exception of synaptic terminals containing acetylcholine,GABA and glutamate which do not typically have the large, densecore vesicles. The most frequent post-synaptic target are dendritesand spinous processes. Less frequently, synaptic contacts arepresent on the cell soma. Chem. Senses 21: 367–376, 1996.     

Immunohistochemical Localization of Laminin and Cytokeratin in Embryonic Alligator Gonads

Gonadal sex differentiation is temperature-dependent in Alligator mississippiensis; testis differentiation occurs in embryos incubated at 33°C and ovary differentiation occurs in embryos incubated at 30°C. Laminin and cytokeratin were examined immunohistochemically in the gonads of alligator embryos incubated at these temperatures. The aim of this study was to determine whether these structural proteins show the same sex-specific expression patterns reported for mammalian embryos, and to assess their usefulness as early markers of gonadal differentiation in species with temperature-dependent sex determination. Laminin delineated enlarged seminiferous cords in differentiating testes from developmental stage 23 to hatching. Laminin distribution was more diffuse and revealed smaller cords of cells in differentiating ovaries. Cytokeratin was also detected in developing gonads of both sexes. Cytokeratin became concentrated in the basal cytoplasm of differentiating Sertoli cells in developing testes. In developing ovaries, prefollicular cells of the ovarian cortex and cell cords in the medulla stained strongly for cytokeratin. Cytokeratin did not show the same basal distribution in female medullary cord cells as seen in the Sertoli cells of testes, however. These sex-specific patterns of laminin and cytokeratin distribution in embryonic alligator gonads may serve as early markers of sexual differentiation.     

Immunohistochemical Localization of Phosphodiesterase 2A in Multiple Mammalian Species

Phosphodiesterases (PDEs) comprise a family of enzymes that regulate the levels of cyclic nucleotides, key second messengers that mediate a diverse array of functions. PDE2A is an evolutionarily conserved cGMP-stimulated cAMP and cGMP PDE. In the present study, the regional and cellular distribution of PDE2A in tissues of rats, mice, cynomolgus monkeys, dogs, and humans was evaluated by immunohistochemistry. A polyclonal antibody directed to the C-terminal portion of PDE2A specifically detected PDE2A by Western blotting and by immunohistochemistry. The pattern of PDE2A immunoreactivity (ir) was consistent across all species. Western blot analysis demonstrated that PDE2A was most abundant in the brain relative to peripheral tissues. PDE2A ir was heterogeneously distributed within brain and was selectively expressed in particular peripheral tissues. In the brain, prominent immunoreactivity was apparent in components of the limbic system, including the isocortex, hippocampus, amygdala, habenula, basal ganglia, and interpeduncular nucleus. Cytoplasmic PDE2A staining was prominent in several peripheral tissues, including the adrenal zona glomerulosa, neurons of enteric ganglia, endothelial cells in all organs, lymphocytes of spleen and lymph nodes, and pituitary. These studies suggest that PDE2A is evolutionarily conserved across mammalian species and support the hypothesis that the enzyme plays a fundamental role in signal transduction. (J Histochem Cytochem 57:933–949, 2009)     

Immunohistochemical Localization of Six Galectin Subtypes in the Mouse Digestive Tract

Galectin, an animal lectin that recognizes β-galactoside of glycoconjugates, is abundant in the gut. This IHC study showed the subtype-specific localization of galectin in the mouse digestive tract. Mucosal epithelium showed region/cell-specific localization of each galectin subtype. Gastric mucous cells exhibited intense immunoreactions for galectin-2 and galectin-4/6 with a limited localization of galectin-3 at the surface of the gastric mucosa. Electron microscopically, galectin-3 immunoreactivity coated indigenous bacteria on the gastric surface mucous cells. Epithelial cells in the small intestine showed characteristic localizations of galectin-2 and galectin-4/6 in the cytoplasm of goblet cells and the baso-lateral membrane of enterocytes in association with maturation, respectively. Galectin-3 expressed only at the villus tips was concentrated at the myosin-rich terminal web of fully matured enterocytes. Epithelial cells of the large intestine contained intense immunoreactions for galectin-3 and galectin-4/6 but not for galectin-2. The stratified squamous epithelium of the forestomach was immunoreactive for galectin-3 and galectin-7, but the basal layer lacked galectin-3 immunoreactivity. Outside the epithelium, only galectin-1 was localized in the connective tissue, smooth muscles, and neuronal cell bodies. The subtype-specific localization of galectin suggests its important roles in host-pathogen interaction and epithelial homeostasis such as membrane polarization and trafficking in the gut. (J Histochem Cytochem 57:41–50, 2009)     

七彩山鸡胃肠道5-羟色胺细胞的免疫组织化学定位

目的 应用5-羟色胺(5-hydroxtryptamine,5-HT)抗血清研究七彩山鸡Phasianus colchicas胃肠道5-HT免疫活性细胞的分布密度和形态,探讨其分布型的成因及细胞形态与功能的关系.方法 免疫组织化学ABC法.结果 5-HT细胞主要分布在十二指肠及其以下部位,腺胃偶见,分布密度近似波浪型,其中以空肠和十二指肠分布密度最高,盲肠和回肠次之,直肠最少.5-HT细胞的形态多样,呈圆形、椭圆形、锥体形等,主要分布于胃肠固有膜、粘膜上皮细胞之间、粘膜上皮细胞基部、腺泡上皮细胞之间.结论 七彩山鸡胃肠道5-HT细胞分布型的形成与各部位消化功能有关,根据其形态,认为七彩山鸡胃肠道5-HT细胞具有内、外分泌两种功能.     

Immunohistochemical Localization of IAA in the Leaf Abscission Zone of Syringa oblata

Freezing sections and immunogold-silver staining were employed to the study on the localization of IAA in petioles of Syringa oblata Lind. At different stages of leaf abscission, the distribution patterns of the silver particles varied in different tissues. In the earlier period of abscission, there were many silver particles in the proximal and distal tissues, but only a few in the abscission zone. The high density of silver particles was found in the phloem of the petiole. The number of silver particles in the abscission zone increase immediately after the protective layer was formed and began to decrease along with the development of the abscission zone. The density of the silver particles became very low when abscission was completed. The formation of protective layer may be the demarcation line of the Stage Ⅰ and Stage Ⅱ during the development of the abscission zone.     

红腹锦鸡胃肠道内分泌细胞的免疫组织化学定位

应用ABC免疫组织化学方法,对红腹锦鸡(Chrysolophus pictus)胃肠道5-羟色胺(5-hydroxtryptamine,5-HT)、胃泌素(gastfin,GAS)、生长抑素(somatostatin,ss)3种内分泌细胞的分布密度和形态进行了观察.结果显示,5-HT细胞在空肠和直肠分布密度最高,回肠和盲肠次之,十二指肠较少,腺胃和肌胃最少;GAS细胞在十二指肠和直肠分布密度最高,其次是空肠和盲肠,腺胃部最低,肌胃则呈免疫阴性;SS细胞数量较少,在直肠、盲肠处分布密度相对高,其次是十二指肠和空肠,腺胃部最低,肌胃则呈免疫阴性.3种内分泌细胞的形态多样,有圆形、椭圆形、锥体形、杆状和不规则形,其中以圆形、椭圆形为主.细胞分布于固有膜、黏膜上皮细胞基部、黏膜上皮细胞之间、腺泡上皮细胞基部或腺泡上皮细胞之间.红腹锦鸡胃肠道内分泌细胞的形态与其内、外分泌功能是相适应的.     

长鬣蜥胃肠道5-羟色胺细胞的免疫组织化学定位

研究长鬣蜥消化道内含有的5一羟色胺（5-HT）内分泌细胞的位置和形态,采用ABC（avidin—biotin—peroxidase complex）免疫组织化学法,应用5-HT特异性抗血清,对其消化道内的5-HT细胞进行免疫化学定位。结果：5-HT细胞从胃部到直肠的胃肠道各段均有分布,细胞密度分布以胃部最高,十二指肠次之,回肠最低。5-HT细胞的形态呈圆形、椭圆形和锥体形等,其中胃体部和直肠部以圆形和椭圆形为主,小肠部以锥体形为主;5-HT细胞广泛分布于上皮基部、上皮细胞之间、腺泡上皮及腺泡之间,有时可见于固有膜内。结论：5-HT细胞的密度分布与其食性、食物组成和生活环境有关,5-HT细胞的形态与其内、外分泌是相适应的。