Wha-Sun Kang

Differential regulation of osteopontin receptors, CD44 and the αv and β3 integrin subunits, in the rat hippocampus following transient forebrain ischemia

We have examined the spatiotemporal regulation of CD44 and the alpha(v)beta(3) integrin subunits, which have been identified as receptors for osteopontin (OPN), in the rat hippocampus following transient forebrain ischemia. Immunoreactivity for CD44 and the integrin subunits, alpha(v) and beta(3), showed characteristic time- and cell-dependent patterns in the ischemic hippocampus. CD44 immunoreactivity was induced at day 1 after reperfusion, reached a peak at day 3, and returned to basal levels by day 7. CD44 was induced in a subset of activated microglial cells within sites of intense neural damage, and the concomitant induction of OPN and CD44 was observed in the same cells in the ischemic hippocampus. In contrast, increased immunoreactivity for alpha(v) and beta(3), which shared overlapping expression patterns in the ischemic hippocampus, occurred in the majority of reactive astrocytes and only a few microglia at day 3 after reperfusion, and was sustained for more than 2 weeks. Immunoreactivity for both integrin subunits colocalized with OPN immunoreactivity in reactive astrocytes, and OPN immunoreactivity was also diffusely localized over the extracellular matrix around the reactive astrocytes. These data indicated that the rapid and transient induction of CD44 and OPN occurred in activated microglia/macrophages, whereas the long-lasting induction of alpha(v) and beta(3) integrin subunits and OPN occurred in reactive astrocytes, suggesting that the multifunctional role of OPN in the ischemic brain may be attributed, in part, to a time- and cell-dependent interaction with CD44 or integrin alpha(v)beta(3).

AII amacrine cells in the mammalian retina show disabled-1 immunoreactivity.

Disabled 1 (Dab1) is an adapter molecule in a signaling pathway, stimulated by Reelin, which controls cell positioning in the developing brain. It has been localized to AII amacrine cells in the mouse and guinea pig retinas. This study was conducted to identify whether Dab1 is commonly localized to AII amacrine cells in the retinas of other mammals. We investigated Dab1‐labeled cells in human, rat, rabbit, and cat retinas in detail by immunocytochemistry with antisera against Dab1. Dab1 immunoreactivity was found in certain populations of amacrine cells, with lobular appendages in the outer half of the inner plexiform layer (IPL) and a bushy, smooth dendritic tree in the inner half of the IPL. Double‐labeling experiments demonstrated that all Dab1‐immunoreactive amacrine cells were immunoreactive to antisera against calretinin or parvalbumin (i.e., other markers for AII amacrine cells in the mammalian retina) and that they made contacts with the axon terminals of the rod bipolar cells in the IPL close to the ganglion cell layer. Furthermore, all Dab1‐labeled amacrine cells showed glycine transporter‐1 immunoreactivity, indicating that they are glycinergic. The peak density was relatively high in the human and rat retinas, moderate in the cat retina, and low in the rabbit retina. Together, these morphological and histochemical observations clearly indicate that Dab1 is commonly localized to AII amacrine cells and that antiserum against Dab1 is a reliable and specific marker for AII amacrine cells of diverse mammals. J. Comp. Neurol. 470:372–381, 2004.

The expression and cellular localization of phospholipase D1 in the rodent retina.

Phospholipase D (PLD) is one of the intracellular signal transduction enzymes and plays an important role in a variety of cellular functions. We investigated the expression and cellular localization of the PLD isozyme PLD1 in the rodent retina. Western blot analysis showed the presence of PLD1 at the protein level in the rat, mouse and guinea pig retinas. PLD1 immunoreactivity was localized in all Müller cells. Thus, PLD1 protein appears to be important in the functions of these cells in the rodent retina.

Light- and electron-microscopic analysis of vasoactive intestinal polypeptide-immunoreactive amacrine cells in the guinea pig retina

Vasoactive intestinal polypeptide (VIP) is a neuroactive substance that is expressed in both nonmammalian and mammalian retinas. This study investigated the morphology and synaptic connections of VIP‐containing neurons in the guinea pig retina by immunocytochemistry, by using antisera against VIP. Specific VIP immunoreactivity was localized to a population of wide‐field and regularly spaced amacrine cells with processes ramifying mainly in strata 1 and 2 of the inner plexiform layer (IPL). Double‐label immunohistochemistry demonstrated that all VIP‐immunoreactive cells possessed γ‐aminobutyric acid immunoreactivity. The synaptic connectivity of VIP‐immunoreactive amacrine cells was identified in the IPL by electron microscopy. The VIP‐labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in strata 1 to 3 of the IPL. The most frequent postsynaptic targets of VIP‐immunoreactive amacrine cells were other amacrine cell processes in strata 1 to 3 of the IPL. Synaptic outputs to bipolar cells were also observed in strata 1 to 3 of the IPL. In addition, ganglion cell dendrites were also postsynaptic to VIP‐immunoreactive neurons in the sublamina a of the IPL. These studies show that one type of VIP‐immunoreactive amacrine cells make contact predominantly with other amacrine cell processes. This finding suggests that VIP‐containing amacrine cells may influence inner retinal circuitry, thus mediating visual processing. J. Comp. Neurol. 445: 325–335, 2002.

Light- and electron-microscopic analysis of neuropeptide Y-immunoreactive amacrine cells in the guinea pig retina

We investigated the morphology and synaptic connections of neuropeptide Y (NPY)-containing neurons in the guinea pig retina by immunocytochemistry, using antisera against NPY. Specific NPY immunoreactivity was localized to a population of wide-field and regularly spaced amacrine cells with processes ramifying mainly in stratum 1 of the inner plexiform layer (IPL). Double-label immunohistochemistry demonstrated that all NPY-immunoreactive cells possessed glutamic acid decarboxylase 65 immunoreactivity. The synaptic connectivity of NPY-immunoreactive amacrine cells was identified in the IPL by electron microscopy. The NPY-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in stratum 1 of the IPL. The most frequent postsynaptic targets of NPY-immunoreactive amacrine cells were other amacrine cell processes. Synaptic outputs to bipolar cells were also observed in a small number of cases. This finding suggests that NPY-containing amacrine cells may influence inner retinal circuitry in stratum 1 of the IPL, thus mediating visual processing.

The absence of the clathrin-dependent endocytosis in rod bipolar cells of the FVB/N mouse retina

The high rate of exocytosis at the ribbon synapses is balanced by following compensatory endocytosis. Unlike conventional synaptic terminals where clathrin-mediated endocytosis (CME) is a predominant mechanism for membrane retrieval, CME is thought to be only a minor mechanism of endocytosis at the retinal ribbon synapses, but CME is present there and it works. We examined the clathrin expression in the FVB/N rd1 mouse, which is an animal model of retinitis pigmentosa. The broadly distributed pattern of clathrin immunoreactivity in the inner plexiform layer was similar in both the control and FVB/N mouse retinas, but the immunoreactive punta within the rod bipolar axon terminals located in the proximal IPL were decreased in number and reduced in size at postnatal days 14 and they came to disappear at postnatal days 21. This preferential decrease of the clathrin expression at ribbon synapses in the rod bipolar cell axon terminals of the FVB/N mouse retina demonstrates another plastic change after photoreceptor degeneration and this suggests that clathrin may be important for normal synaptic function at the rod bipolar ribbon synapses in the mammalian retina.

Cell-type specific expression of vanilloid receptor 1 in the taste cells of rat circumvallate papillae

The present study demonstrates the first-ever characterization of cell types that express the vanilloid receptor 1 (VR1) in the taste buds of rat circumvallate papillae. We performed electron microscopy to identify the subcellular location. The VR1 immunoreactivity was associated with the endoplasmic reticulum, cytoplasmic vesicles, and plasma membrane of taste cells. These results demonstrate the localization of the VR1 in membranous structures of the taste cells. We used double immunofluorescence histochemistry with taste cell type-specific markers to identify the cell types that express the VR1. The VR1 was detected in all functional taste cell types (Type I, Type II, and Type III cells). Together, our data suggest that the VR1 might play different roles according to the cell types within a taste bud.

Regulatory expression and cellular localization of doublecortin in the rat retina following ischemia-reperfusion injury

Doublecortin (DCX) is microtubule-associated protein and is required for neuronal migration, differentiation and plasticity. In the retina, it is highly expressed between embryonic day 18 (E18) and E20, and is poorly expressed postnatally. In this study, we investigated the expression and cellular localization of DCX in the rat retina following ischemia induced by transiently increasing the intraocular pressure. While DCX immunoreactivity in control retinas was restricted to the outer border of the inner nuclear layer, it appeared in horizontal cell somata and processes in affected retinas. Quantitative evaluation by immunoblotting confirmed that DCX expression continuously increased after ischemia-reperfusion and showed 370% of control protein levels at 4 weeks after ischemic insult. These results suggest that the DCX in horizontal cells might play a role in neurite remodeling or modulating other neurons in ischemic rat retinas.