The immunolocalization of cluster of differentiation 31, phalloidin and alpha smooth muscle actin on vascular network of normal and ischemic rat brain

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.

Melanocortin Derivatives Induced Vascularization and Neuroglial Proliferation in the Rat Brain under Conditions of Cerebral Ischemia

“…CD31 antibodies, widely used as an endothelial marker, are used to immunostain existing and developing brain blood vessels under both physiological and pathological conditions [ 66 ]. The results of our immunohistochemical studies using the endothelial marker CD31 showed a significant increase in the intact vessels number of the microcirculatory bed in the perifocal region of the infarction when the studied peptides were used ( Figure 5 ).…”

Activin A, a Novel Chemokine, Induces Mouse NK Cell Migration via AKT and Calcium Signaling

“…Cell motility is also closely related to the polarization distribution of cytoskeletal proteins [16,24,25]. Phalloidin has a high affinity for actin and can selectively bind filamentous actin (F-actin) without binding monomeric actin (G-actin) [26,27]. This study used green fluorescent phalloidin to examine the expression and distribution of F-actin in mouse NK cells during cell migration in microfluidic chips.…”

“…Compared to the control group, the expression of β-catenin and vimentin was enhanced in activin A-treated NK cells, and the expression of MMP2, which is involved in ECM degradation, was also significantly increased (Figure 4B). actin (F-actin) without binding monomeric actin (G-actin) [26,27]. This study used green fluorescent phalloidin to examine the expression and distribution of F-actin in mouse NK cells during cell migration in microfluidic chips.…”