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1.
Synopsis Carotid bodies of adult albino rats were examined using the formaldehyde-induced fluorescence (FIF) method for the demonstration of fluorogenic monaomines and staining with I% Toluidine Blue for morphological observations.In the carotid body of normal controls, most glomus (principal or type I) cells exhibited a FIF presumably due to catecholamines. The intensity of the fluorescence was weak in most cells, while some glomus cells were non-fluorescent and others exhibited a moderate or intense FIF. The sustentacular (satellite, supporting or type II) cells were essentially non-fluorescent.One week after the administration of a single intraperitoneal injection of the long-acting glucocorticoid 6-methylprednisolone sypionate (400 mg/kg) or after seven intraperitoneal injections of the water-soluble glucocorticoid hydrocortisone sodium succinate (40 mg/kg daily for a week), a distinct increase was observed in the FIF of the glomus cells. No non-fluorescent glomus cells were observed after treatment with either glucocorticoid, and the intensity of most fluorescent glomus cells was moderate or intense.It is concluded that glucocorticoids cause an increased storage of catecholamines in the glomus cells of the carotid body of the adult rat, an observation of interest in view of the fact that such changes due to glucocorticoids have as yet been reported only in catecholamine-storing cells of newborn rats. 相似文献
2.
Zhu XY Bentley MD Chade AR Ritman EL Lerman A Lerman LO 《American journal of physiology. Heart and circulatory physiology》2007,293(3):H1997-H2003
Changes in the structure of the artery wall commence shortly after exposure to cardiovascular risk factors, such as hypercholesterolemia (HC), but may be difficult to detect. The ability to study vascular wall structure could be helpful in evaluation of the factors that instigate atherosclerosis and its pathomechanisms. The present study tested the hypothesis that early morphological changes in coronary arteries of hypercholesterolemic (HC) pigs can be detected using the novel X-ray contrast agent OsO(4) and three-dimensional micro-computed tomography (CT). Two groups of pigs were studied after they were fed a normal or an HC (2% cholesterol) diet for 12 wk. Hearts were harvested, coronary arteries were injected with 1% OsO(4) solution, and cardiac samples (6-mum-thick) were scanned by micro-CT. Layers of the epicardial coronary artery wall, early lesions, and perivascular OsO(4) accumulation were determined. Leakage of OsO(4) from myocardial microvessels was used to assess vascular permeability, which was correlated with immunoreactivity of vascular endothelial growth factor in corresponding histological cross sections. OsO(4) enhanced the visualization of coronary artery wall layers and facilitated detection of early lesions in HC in longitudinal tomographic sections of vascular segments. Increased density of perivascular OsO(4) in HC was correlated with increased vascular endothelial growth factor expression and suggested increased microvascular permeability. The use of OsO(4) as a contrast agent in micro-CT allows three-dimensional visualization of coronary artery wall structure, early lesion formation, and changes in vascular permeability. Therefore, this technique can be a useful tool in atherosclerosis research. 相似文献
3.
Summary In an attempt to identify pancreatic islet cells emitting formaldehyde-induced fluorescence (FIF), the pancreatic islets of the domestic fowl were studied by combined fluorescence, ultrastructural, silver-impregnation and immunohistochemical methods in the same section or in consecutive semi-thin and ultra-thin sections. The results indicate that islet cells emitting intense FIF exhibit a strongly argyrophil reaction with the Grimelius' silver method and also immunohistochemical reaction with anti-glucagon serum, but not with anti-5-HT serum. Therefore, the fowl islet A cell, a peptide hormone-producing cell, stores simultaneously catecholamine as biogenic amine. The islet B and D cells did not display any FIF, any argyrophil reaction with the Grimelius' silver method, or any immunoreactivity with anti-glucagon or anti-5-HT sera. The fluorescent but non-argyrophil cells dispersed in the exocrine acinus may well be PP cells. 相似文献
4.
Dr. Tohru Watanabe Hirotaka Chikazawa Narong Chungsamarnyart Toshitake Fujioka Junzo Yamada 《Cell and tissue research》1987,247(1):25-32
Summary In an attempt to identify duodenal endocrine cells emitting formaldehyde-induced fluorescence (FIF), chicken duodena were studied by combined fluorescence, ultrastructural, silver impregnation and immunohistochemical methods in the same or consecutive sections. Our results show that: (1) Almost all the cells emitting yellow fluorescence by both the Falck-Hillarp and the Furness methods exhibit an immunohistochemical reaction with serotonin (5-HT) antiserum. (2) Almost all cells radiating yellow fluorescence by the Furness method stain with toluidine blue in Epon-embedded sections but, by high-voltage electron microscopy, can be subdivided into two types of cell containing either small round or polymorphous types of granules. (3) In the sections from which resin had been removed, all the cells emitting yellow FIF show argentaffinity by the Singh method, but not all cells display argyrophilia with the Grimelius method. (4) Cells exhibiting both argyrophil and argentaffin reactions in deresined serial sections are also separated into two types of cell, containing either small spherical or polymorphous types of granules by conventional electron microscopy in thin sections. Therefore, chicken enterochromaffin cells emit yellow FIF, store 5-HT, show both argentaffinity and argyrophilia, but are ultrastructurally classified into two types of granule-containing cells which may be related to polypeptides coexisting with 5-HT. 相似文献
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6.
According to the earlier concept, the paraganglia of man are believed to degenerate during the first postnatal years after their dominance during the fetal period. Clinical case reports on persisting paraganglia led us to extensive exploration of surgical material obtained from urological and gynecological surgery. The formaldehyde induced fluorescence (FIF) was used for tracing the catecholamine containing tissues. The fluorescence intensities were recorded with a Leitz MPV 2 microspectrophotometer. Solitary, small paraganglia were found in all patients studied. They were especially frequent in the walls of the urinary bladder and in the connective tissue surrounding the urogenital organs. The intensity of the fluorescence was comparable to pharmacological standard of 10(-2) M noradrenaline and at the same level as the FIF of human fetal paraganglia. All cells of the paraganglionic clusters exhibited FIF and no signs of degeneration could be observed. It is suggested that the paraganglia of man do not degenerate postnatally but persist as a remarcable catecholamine reservoir, which might be of physiological importance. 相似文献
7.
Paraganglia in the urogenital tract of man 总被引:1,自引:0,他引:1
A. Hervonen A. Vaalasti T. Vaalasti M. Partanen L. Kanerva 《Histochemistry and cell biology》1976,48(4):307-313
Summary According to the earlier concept, the paraganglia of man are believed to degenerate during the first postnatal years after their dominance during the fetal period. Clinical case reports on persisting paraganglia led us to extensive exploration of surgical material obtained from urological and gynecological surgery. The formaldehyde induced fluorescence (FIF) was used for tracing the catecholamine containing tissues. The fluorescence intensities were recorded with a Lietz MPV 2 microspectrophotometer.Solitary, small paraganglia were found in all patients studied. They were expecially frequent in the walls of the urinary bladder and in the connective tissue surrounding the urogenital organs. The intensity of the fluorescence was comparable to pharmacological standard of 10–2 M noradrenaline and at the same level as the FIF of human fetal paraganglia. All cells of the paraganglionic clusters exhibited FIF and no signs of degeneration could be observed.It is suggested that the paraganglia of man do not degenerate postnatally but persist as a remarcable catecholamine reservoir, which might be of physiological importance. 相似文献
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9.
D. F. Newgreen I. J. Allan H. M. Young B. R. Southwell 《Development genes and evolution》1981,190(6):320-330
Summary Catecholamines (CA) were localized in stage 11–34 domestic fowl embryos by the formaldehyde-induced fluorescence (FIF) method after exposure in vivo or in vitro to CA (noradrenaline or -methylnoradrenaline), or the CA precursorl-DOPA. The effects of drugs known to alter CA metabolism in the adult were also investigated.Negligible FIF was observed in embryos which had not been exposed to CA. After CA loading, FIF could be seen in the neural tube and in non-neural tissues such as the notochord and gut mesenchyme and to a lesser degree in suprarenal area tissue, liver endothelium, sclerotome, and myotome. This FIF was inhibited by desmethylimipramine, a blocker of adult neuronal CA uptake (Uptake1), but not by corticosterone, a blocker of adult extraneuronal CA uptake (Uptake2). The notochord, dorsal pancreas and some blood cells were fluorescent afterl-DOPA loading, and this FIF could be greatly diminished by the DOPA decarboxylase inhibitor RO4-4602.The pattern of FIF in the axial structures (neural tube and notochord) correlated with axial flexure in both position and time, and the intensity of fluorescence was strongest cranially and caudally, where flexure is most pronounced. The FIF in gut mesenchyme cells was closely related to the movement of the intestinal protals during early gut tube formation, and to the regions of the developing intestine that undergo intense morphogenesis during their early formation. 相似文献
10.
S Yamaguchi M Yokoyama T Iida M Okai O Tanaka A Takimoto 《Plant & cell physiology》2001,42(11):1201-1209
A stress-induced fatty acid [FIF; 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid] incubated with (-)-norepinephrine (NE) strongly induces flower formation in Lemna paucicostata [Yokoyama et al. (2000), Plant Cell Physiol. 41: 110). The increase of flower-inducing activity was well correlated with the decrease in FIF in the incubation mixture, and the reaction proceeded rapidly at higher pH. We detected small amounts of many active components in the mixture after incubation by HPLC analysis. In this study, two major components, named FN1 and FN2, of the reaction mixture were isolated, and their absolute stereostructures were determined. FN1 showed a strong flower-inducing activity and was identified as a tricyclic alpha-ketol fatty acid, 9(R)-11-[(2'R,8'R,10'S,11'S)-2',8'-dihydroxy-7'-oxo-11'-[(Z)-2-pentenyl]-9'-oxa-4'-azatricyclo[6.3.1.0(1.5)]dodec- 5'en-10'-yl]-9-hydroxy-10-oxoundecanoic acid [corrected]. FN2, the C-9 epimer of FN1, showed no flower-inducing activity. The absolute stereostructure of FIF was also determined by a modification of Mosher's method. The 9-hydroxyl group was found to be predominantly 9R, with an enantiomeric excess of 40% (70% 9R and 30% 9S). FN1 was derived from 9R-type FIF and FN2 from 9S-type FIF. Various catecholamines and related substances were investigated for the ability to develop flower-inducing activity upon incubation with FIF. The essential structures were catechol and ethylamine groups (dopamine). 相似文献
11.
1--The innervation of the liver and gallbladder of Rana ridibunda has been studied by the following methods: (a) demonstration of cholinesterase activity; (b) FIF method for catecholamines; (c) immunohistochemistry for VIP and (d) electron microscopy. 2--The hepatocytes are arranged in regular rows of hepatic cords, very little connective tissue is distributed in the parenchyma, the innervation being restricted to the big branches of blood vessels. 3--Well defined cholinergic and adrenergic plexuses surround the hepatic arteries, portal veins and biliary ducts. The VIPergic innervation is scarce in the liver but a richly branched plexus spreads in the wall of the gallbladder. 4--Cholinesterase-positive cells are widely distributed accompanying the nerve trunks of the gallbladder. The innervation distribution is prominent in the portion of the gallbladder next to the hepatic hilus. 5--A population of melanin-storing cells besides free melanin granules are present in the liver parenchyma and are prominent in the gallbladder where the melanocytes are disposed in close contact with blood vessels and nerve structures. We have observed that the number of these visceral melanocytes considerably increases in winter, particularly in the liver. 相似文献
12.
A growing body of evidence suggests that the angiotensin II fragments, Ang(1-7) and Ang(3-8), have a vasoactive role, however ACE2, the enzyme that produces Ang(1-7), or AT4R, the receptor that binds Ang (3-8), have yet been simultaneously localised in both normal and diseased human conduit blood vessels. We sought to determine the immunohistochemical distribution of ACE2 and the AT4R in human internal mammary and radial arteries from patients undergoing coronary artery bypass surgery. We found that ACE2 positive cells were abundant in both normal and diseased vessels, being present in neo-intima and in media. ACE2 positive immunoreactivity was not present in the endothelial layer of the conduit vessels, but was clearly evident in small newly formed angiogenic vessels as well as the vaso vasorum. Endothelial AT4R immunoreactivity were rarely observed in either normal and diseased arteries, but AT4R positive cells were observed adjacent to the internal elastic lamine in the internal mammary artery, in the neo-intima of radial arteries, as well as in the media of both internal mammary artery and radial artery. AT4R was abundant in vaso vasorum and within small angiogenic vessels. Both AT4R and ACE2 co-localised with smooth muscle cell alpha actin. This study identifies smooth muscle cell alpha actin positive ACE2 and AT4R in human blood vessels as well as in angiogenic vessels, indicating a possible role for these enzymes in pathological disease. 相似文献
13.
Adrenergic innervation of the human gall bladder was studied using two specific fluorescence histochemical methods. Blue-green fluorescing varicose nerves were scarce and mostly followed the course of blood vessels as typical perivascular plexuses. However, some adrenergic nerves not associated with the vessels were occasionally seen, as well as structures suggestive of a pericellular arrangement of varicose adrenergic nerve terminals on non-fluorescing ganglion cells. A few enterochromaffin cells were seen in the epithelial lining, also in the deep invaginations obviously representing the Aschoff-Rokitansky sinuses. Occasionally, small rounded cells with a rounded, relatively large nucleus, and exhibiting a weak yellow-green to blue-green granular cytoplasmic fluorescence, were observed in the wall of the gall bladder. The possible functional and evolutionary significance of these neural and endocrine elements was discussed against the data on physiological and pharmacological studies obtained from the literature. It was concluded that their significance is, in all probability, secondary to the influence of the intestinal polypeptide hormones, vagal innervation and circulating catecholamines upon the normal function of the gall bladder. The glyoxylic acid-induced fluorescence histochemical method was found to be superior to the conventional formaldehyde technique in studies on human tissue. 相似文献
14.
Summary Adrenergic innervation of the human gall bladder was studied using two specific fluorescence histochemical methods. Blue-green fluorescing varicose nerves were scarce and mostly followed the course of blood vessels as typical perivascular plexuses. However, some adrenergic nerves not associated with the vessels were occasionally seen, as well as structures suggestive of a pericellular arrangement of varicose adrenergic nerve terminals on non-fluorescing ganglion cells. A few enterochromaffin cells were seen in the epithelial lining, also in the deep invaginations obviously representing the Aschoff-Rokitansky sinuses. Occasionally, small rounded cells with a rounded, relatively large nucleus, and exhibiting a weak yellow-green to blue-green granular cytoplasmic fluorescence, were observed in the wall of the gall bladder. The possible functional and evolutionary significance of these neural and endocrine elements was discussed against the data on physiological and pharmacological studies obtained from the literature. It was concluded that their significance is, in all probability, secondary to the influence of the intestinal polypeptide hormones, vagal innervation and circulating catecholamines upon the normal function of the gall bladder. The glyoxylic acid-induced fluorescence histochemical method was found to be superior to the conventional formaldehyde technique in studies on human tissue. 相似文献
15.
Adult male rats were exposed to toluene in short-term exposure by inhalation for 48 h (2000 ppm, continuously), and in long-term inhalation for 3 months (1000 ppm, 8 h daily). The formaldehyde-induced fluorescence (FIF) technique for histochemical demonstration of catecholamines (CA) was used to detect changes in the catecholamine stores. The concentration of CA in the sympathetic neurons of superior cervical ganglia and adrenal medulla was measured by the FIF technique combined with microfluorimetry. The short-term toluene exposure induced a statistically significant reduction of CA contents in sympathetic neurons. After long-term exposure, no change in the CA level could be demonstrated either in sympathetic ganglion or adrenal medulla. In electron microscopic studies no clear pathological changes were detected after toluene exposure. 相似文献
16.
Expression of serotonin receptor mRNAs in blood vessels 总被引:17,自引:0,他引:17
Using RT-PCR we distinguished mRNAs for all known G-protein coupled serotonin receptors expressed in various rat and porcine blood vessels. Nearly all vessels expressed 5HT1
β, 5-HT2A, 5-HT2B, 5-HT4, and 5-Ht7 receptor mRNA to different extents. New splice variants of the porcine 5-HT4 receptor were observed. Similar PCR assays were performed with endothelial and smooth muscle cells from human pulmonary artery, aorta, and with endothelial cells from human coronary artery and umbilical vein. All endothelial cells expressed 5-HT1
β, 5-HT2b, and 5-HT4 receptor mRNA, whereas in smooth muscle cells 5-HT1
β, 5-HT2A, 5-HT7, and in some experiments 5-HT2B receptor mRNA were found. A model for the regulation of vascular tone by different 5-HT receptors is proposed. 相似文献
17.
The innervation of the liver and gallbladder of the lizard Podarcis hispanica has been studied by the following methods: a) demonstration of cholinesterase activity; b) FIF method for catecholamines; and c) immunohistochemistry for VIP. The hepatic parenchyma of the reptile's liver show hepatocytes arranged in regular rows of hepatic cords, the portal triad being typical of higher vertebrates (birds and mammals). Nerve fibers are found in the scarce connective tissue distributed among the hepatocytes. The innervation is restricted to the big branches of blood vessels and biliary ducts. It is represented by cholinergic, noradrenergic and VIPergic fibers. The gallbladder shows a well developed cholinergic plexus with pyramidal cells in the interconnection points of the fiber network. The noradrenergic and VIPergic plexuses are also more widely distributed in the gallbladder than in the liver. 相似文献
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19.
We recently reported a new lymphokine activity that affects fibroblasts by inhibiting their spontaneous migration. Human fibroblast migration inhibitory factor (FIF) obtained from concanavalin A (Con A)-stimulated human lymphocytes was characterized by Sephadex gel filtration and by enzyme treatment. FIF was found to be stable at 56 degrees C for 15 min but destroyed at 80 degrees C or at pH lower than 5. Gel filtration revealed two peaks of FIF activity 15,000 and at 34,000 Da. FIF activity was lost following treatment with trypsin, chymotrypsin, and neuraminidase and FIF could not be generated in the presence of inhibitors of glycosylation, suggesting that the molecule was a glycoprotein. FIF could be removed by adsorption to human fibroblasts but not to PMN, monocytes, or red blood cells. Further studies were carried out to investigate the role of sugars in the interaction of FIF with the target cells. Human FIF activity was significantly reduced in the presence of several sugars including alpha-methyl-D-mannoside, L-xylose, N-acetyl-D-glucosamine, D-mannose, L-rhamnose but not L-fucose. Preincubation of human fibroblasts with alpha-methyl-D-mannoside prevented their response to FIF. In contrast, pretreatment of fibroblasts with mannosidase had no effect, suggesting that alpha-methyl-D-mannoside was an essential component of the FIF molecule recognized by the FIF receptor on fibroblasts. 相似文献
20.
Parathyroid hormone-related peptide (PTHrP) is known to be a strong vasorelaxant peptide. The mechanisms by which PTHrP reduces the coronary resistance of the rat heart have not been worked out but seem to be independent of the classical PTH/PTHrP receptor-mediated, cAMP-dependent effect. In this study we hypothesized that PTHrP reduces the coronary resistance of the rat heart via endothelial cell hyperpolarization. Isolated microvascular endothelial cells from rat heart were incubated with PTHrP(1-36), and changes in the membrane potential were recorded via DiBAC fluorescence. Cells exposed to PTHrP showed a hyperpolarization of approximately 7mV. In the isolated Langendorff preparation, PTHrP-dependent vasodilatation of l-nitro-arginine-exposed hearts was abolished under depolarizing conditions (high potassium). Denudation of the endothelial cell layer significantly impaired the vasodilatory effect of PTHrP. In the presence of H89 (a cAMP/protein kinase A pathway antagonist) and indomethacin (a cyclooxygenase inhibitor), PTHrP dilated the vessels. In conclusion, PTHrP exerted a nitric oxide-independent vasodilatory effect that depends on endothelial cell hyperpolarization. 相似文献