Improved Penile Histology by Phalloidin Stain: Circular and Longitudinal Cavernous Smooth Muscles, Dual-endothelium Arteries, and Erectile Dysfunction-associated Changes
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Abstract Cluster of differentiation 31 (CD31), phalloidin and alpha smooth muscle actin (α-SMA) have been widely applied to label the cerebral blood vessels in the past years. Although CD31 is mainly used as endothelial marker in determining the cerebral capillaries, it seems likely that its labeling efficiency is closely correlated with the antibodies from the polyclonal or monoclonal one, as well as the conditions of blood vessels. In order to test this phenomenon, we compared the labeling characteristics of goat polyclonal anti-CD31 (gP-CD31) and mouse monoclonal anti-CD31 (mM-CD31) with those of phalloidin and α-SMA on the rat brain in health and ischemia/reperfusion (I/R) with the middle cerebral artery occlusion. By multiple immunofluorescence staining, it was found that gP-CD31 labeling expressed extensively on the cerebral capillaries forming the vascular networks on the normal and ischemic regions, but mM-CD31 labeling mainly presented on the capillaries in the ischemic region. In contrast to the vascular labeling with gP-CD31, phalloidin and α-SMA were mainly expressed on the wall of cortical penetrating arteries, and less on that of capillaries. By three-dimensional reconstruction analysis, it was clearly shown that gP-CD31 labeling was mainly located on the lumen side of vascular wall and was surrounded by phalloidin labeling and α-SMA labeling. These results indicate that gP-CD31 is more sensitive than mM-CD31 for labeling the cerebral vasculature, and is highly compatible with phalloidin and α-SMA for evaluating the cerebral vascular networks under the physiological and pathological conditions.
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Phalloidin produces marked protrusions on the surface of isolated hepatocytes prepared from rat livers. This typical in-vitro effect of the mushroom toxin is prevented or inhibited by preincubation with disilybin. The antagonistic effect depends on the concentration of disilybin and might be caused by an inhibition of phalloidin binding.
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Abstract Cluster of differentiation 31 (CD31), phalloidin and alpha smooth muscle actin (α-SMA) has been widely applied for labeling the cerebral blood vessels in the past years. CD31 is mainly used as endothelial marker in determining the cerebral capillaries in the past years. However, it seems likely that its labeling efficiency is closely correlated with the antibodies from the polyclonal or monoclonal one, as well as the conditions of blood vessels. In order to test this phenomenon, we firstly compare the labeling characteristics of goat polyclonal anti-CD31 (gP-CD31) and mouse monoclonal anti-CD31 (mM-CD31) on the rat brain in health and ischemic/reperfusion (I/R) with the middle cerebral artery occlusion. By multiple immunofluorescence staining and three-dimensional reconstruction techniques, it was found that gP-CD31 labeling expressed extensively on the cerebral capillaries in the normal and ischemic regions, but mM-CD31 labeling mainly presented on the capillaries in the ischemic region. In contrast to the vascular labeling with phalloidin andα-SMA, gP-CD31 labeling located on the lumen side of vascular wall and surrounded by phalloidin and α-SMA labeling. These results indicate that gP-CD31 is expressed more sensitively than mM-CD31 on the cerebral vasculature, and highly compatible with phalloidin and α-SMA for insight into the cerebral vascular network in a three-dimensional view under the physiological and pathological conditions.
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An attempt has been made to identify molecular markers of intrahepatic cholestasis in mice employing phalloidin as a cholestatic agent. Phalloidin was administered to BALB/c mice at three predetermined dose: 250 microg/kg, 500 microg/kg, and 1 mg/kg for 1, 3, and 7 days. Liver function was estimated to confirm cholestasis. Histopathological observations on liver were also made to confirm liver injury. Phalloidin at 1 mg/kg for 7 days was found to induce cholestasis. Therefore gene expression studies were confined to this group only. A total of 88 genes were found to be affected by phalloidin. These were the genes associated with cytoskeleton regulation as well as tight junction, focal adhesion, and ATP-binding cassette transporters. Such proteins obstruct the removal of bile components from hepatocytes to the bile canaliculus or blood. Phalloidin treatment did not affect the proteins responsible for cell maintenance or death. The authors show that phalloidin-induced intrahepatic cholestasis is manifested by disturbing the cytoskeleton. The set of genes up-regulated by phalloidin can be considered as molecular markers of intrahepatic cholestasis. The observations are further expected to be helpful in the management of cholestatic pharmaceuticals and associated problems of liver diseases in humans.
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Objective To seek a appropriate diagnostic method for condyloma acuminatum. Methods 48 cases of condyloma acuminatum were used,serial sectioned and stained with HE,in situ hybrihistochemistry(ISHC) and immunohistochemistry(IHC) respectively,assessed under microscopy and was analysed statistically.Results 30 IHC positive cases were all positive by ISHC and IHC stain,13 cases(72.2%) in 18 IHC negative cases were positive by ISHC stain,5 cases (27.2%) were positive by repeated IHC stain.In the serial sections,ISHC positive cells were more than IHC positive cells. Conclusion HPV ISCH stain is higher special than that of IHC stain,ISCH method is same as IHC,may be widely used in clinical pathology.
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Condyloma Acuminatum
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