Heat shock protein 27 upregulation and phosphorylation in rat experimental autoimmune encephalomyelitis.

Following stress or inflammation, the 27-kDa heat shock protein (HSP27) is induced in various cell types, where it promotes cell survival and inhibits inflammatory reactions. We examined the expression of HSP27 and phosphorylated HSP27 (p-HSP27) in the spinal cords of Lewis rats with experimental autoimmune encephalomyelitis (EAE). Western blotting analysis revealed low levels of HSP27 and p-HSP27 in the normal spinal cords and significantly higher levels in EAE-affected spinal cords.

Immunohistochemical study of cathepsin D in the spinal cords of rats with clip compression injury.

This study examined the temporal expression of cathepsin D protein and its cellular localization in the spinal cords of rats after a clip compression injury to determine the involvement of cathepsin D in spinal cord injury (SCI). Western blot analysis showed a significant increase in the approximately 31-kDa active form of cathepsin D on days 4 and 7 after the SCI, while the level of the approximately 44-kDa inactive form remained relatively unchanged. Immunohistochemistry revealed cathepsin D with constitutive localization in most neurons and some gliocytes in the normal spinal cord to be intensely immuno-detected primarily in CD68-positive activated macrophages/microglia in the SCI lesions.

Strain-dependent Differences of Locomotor Activity and Hippocampus-dependent Learning and Memory in Mice.

The behavioral phenotypes of out-bred ICR mice were compared with those of in-bred C57BL/6 and BALB/c mice. In particular, this study examined the locomotor activity and two forms of hippocampus-dependent learning paradigms, passive avoidance and object recognition memory. The basal open-field activity of the ICR strain was greater than that of the C57BL/6 and BALB/c strains.

Transient expression of phospholipase D1 during heart development in rats.

The expression of phospholipase D (PLD) isozymes was examined in the hearts of rats at different stages of development. Immunoprecipitation and Western blot analysis revealed weak PLD1 expression in the hearts of day 17 embryos. The level of PLD1 protein increased transiently 0 and 3 days postpartum, and declined gradually beginning 7 days after birth.

Activation of Akt/protein kinase B and extracellular signal-regulated kinase in rats with acute experimental testicular torsion.

The Akt/protein kinase B (PKB) and extracellular signal-regulated kinase (ERK) pathways are involved in cell survival. This study examined the temporal profiles and localization of Akt/PKB and ERK1/2 activation in rat testis after ischemia/reperfusion (I/R). Testicular tissue was collected from normal control rats and rats exposed to reperfusion for 6, 24, and 48 hr after ischemic injury; the tissues were analyzed via Western blotting and immunohistochemistry.

The expression and cellular localization of phospholipase D isozymes in the developing mouse testis.

To examine the involvement of phospholipase D (PLD) isozymes in postnatal testis development, the expression of PLD1 and PLD2 was examined in the mouse testis at postnatal weeks 1, 2, 4, and 8 using Western blot analysis and immunohistochemistry. The expression of both PLD1 and PLD2 increased gradually with development from postnatal week 1 to 8. Immunohistochemically, PLD immunoreactivity was detected in some germ cells in the testis and interstitial Leydig cells at postnatal week 1.

Increased phosphorylation of cyclic AMP response element-binding protein in the spinal cord of Lewis rats with experimental autoimmune encephalomyelitis.

To investigate whether the phosphorylation of cyclic AMP response element-binding protein (CREB) is implicated in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), the change in the level of CREB phosphorylation was analyzed in the spinal cord of Lewis rats with EAE. Western blot analysis showed that the phosphorylation of CREB in the spinal cord of rats increased significantly at the peak stage of EAE compared with the controls (p<0.05) and declined significantly in the recovery stage (p<0.

Expression of nitric oxide synthase isoforms in the porcine ovary during follicular development.

The expression of nitric oxide synthase (NOS) isoforms in the ovaries of pigs was examined to study the involvement of nitric oxide, a product of NOS activity, in the function of the ovary. Western blot analysis detected three types of NOS in the ovary, including constitutive neuronal NOS (nNOS), endothelial NOS (eNOS) and inducible NOS (iNOS); eNOS immunoreactivity was more intense compared with that of iNOS or nNOS. Immunohistochemical studies demonstrated the presence of nNOS and eNOS in the surface epithelium, stroma, oocytes, thecal cells, and endothelial cells of blood vessels.

Activation of p38 mitogen-activated protein kinase in the early and peak phases of autoimmune neuritis in rat sciatic nerves.

To examine the involvement of p38 mitogen-activated protein kinase (MAPK) in autoimmune disorders of the peripheral nerve system, we analyzed the phosphorylation of p38 MAPK protein in the sciatic nerves of Lewis rats with experimental autoimmune neuritis (EAN). Western blot analysis showed that phosphorylated p38 (p-p38) MAPK protein was significantly increased in the sciatic nerves of rats in the early and peak phases of EAN, and declined gradually thereafter. Immunohistochemistry showed that p-p38 MAPK levels were increased in the infiltrating inflammatory cells, including T cells and macrophages, as well as in blood vessels and some Schwann cells in EAN-affected sciatic nerves, as compared to the sciatic nerves of controls.

Upregulation of osteopontin in Schwann cells of the sciatic nerves of Lewis rats with experimental autoimmune neuritis.

We examined the expression of osteopontin (OPN) in the sciatic nerves of rats with experimental autoimmune neuritis (EAN), using immunohistochemistry and immunoblotting, to study its involvement in the pathogenesis of autoimmune peripheral nervous system diseases. Constitutive OPN expression was detected in some Schwann cells; expression was increased after immunization with adjuvant alone. At day 14 after induction of EAN, many Schwann cells had a granular pattern of immunoreactivity, whereas very few inflammatory cells were OPN-positive.

Activation of extracellular signal-regulated kinases in the sciatic nerves of rats with experimental autoimmune neuritis.

To investigate whether the phosphorylation of extracellular signal-regulated kinase (ERK) is involved in autoimmune injury of the peripheral nervous system (PNS), the expression of phosphorylated ERK (p-ERK) was analyzed in experimental autoimmune neuritis (EAN) in rats. Western blot analysis showed that the level of p-ERK was increased significantly in the sciatic nerves of rats on days 14 (p<0.05) and 24 (p<0.

Increased expression of phospholipase D in the heart with experimental autoimmune myocarditis in Lewis rats.

The expression of phospholipase D (PLD) in the hearts of rats with experimental autoimmune myocarditis (EAM) was studied to elucidate the functional role of PLD in the pathogenesis of EAM. Western blot analysis showed that the level of the PLD1 isoform was significantly increased in the hearts of rats with EAM on days 14, 17 and 21 postimmunization (pi) (P < 0.01; control vs EAM at 14 pi, 17 pi and 21 pi).

Involvement of cyclooxygenase-1 and -2 in the sciatic nerve of rats with experimental autoimmune neuritis.

The expression of both cyclooxygenase (COX)-1 and COX-2, which are representative enzymes in prostaglandin synthesis, was evaluated in the sciatic nerve of rats with experimental autoimmune neuritis (EAN). Western blot analysis showed that both COX-1 and COX-2 were significantly increased in the sciatic nerve at the peak stage of EAN and declined during the recovery stage. Vascular endothelial cells in normal sciatic nerves immunostained for both COX-1 and COX-2.

Histochemistry of six lectins in the tissues of the flat fish Paralichthys olivaceus.

Lectins are glycoproteins that specifically bind carbohydrate structures and may participate in the biodefense mechanisms of fish. In this study, the binding of three lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA) and Ulex europaeus (UEA-I) were studied in the gill, liver, intestine, kidney, heart, and spleen of the flat fish Paralichthys olivaceus. DBA was detected in intestinal mucous cells, as well as in gill epithelial and mucous cells.

Immunohistochemical localization of Bcl-2 in the spinal cords of rats with experimental autoimmune encephalomyelitis.

We examined the localization of the anti-apoptotic molecule Bcl-2 in the spinal cords of Lewis rats with experimental autoimmune encephalomyelitis (EAE). Western blot analysis showed that Bcl-2 was constitutively expressed in normal spinal cords, and weakly increased in response to complete Freund's adjuvant(CFA) immunization. In EAE, with infiltration of inflammatory cells into spinal cords, Bcl-2 declined during the peak stage and further decreased during the recovery stage.

Histochemical detection of glycoconjugates in the male reproductive system of the horse.

Lectins are glycoproteins of plant and animal origin that have the ability to bind specific carbohydrate residues of cell glycoconjugates, particularly in terminal positions. In this study, the binding of lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I), was studied in the reproductive systems of male thoroughbred horses.DBA was detected in the stereocilia of the caput and corpus epididymis, and in the vas deferens.

Histochemical characterization of the lectin-binding sites in the equine vomeronasal organ.

The binding specificities of various lectins, such as the Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), and the Bandeiraea simplicifolia BS-1 (Isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I) lectins, were studied in the vomeronasal organ of the horse. The microvilli of the vomeronasal sensory epithelium were positive for DBA, SBA, Isolectin B4, WGA, PNA, and UEA-I. The receptor cells showed intense reactivity for DBA and WGA.

Embryonic intermediate filaments, nestin and vimentin, expression in the spinal cords of rats with experimental autoimmune encephalomyelitis.

Intermediate filaments, including nestin and vimentin, are found in specific cell types in central nervous system (CNS) tissues, particularly immature glial cells and multipotent progenitor cells. In the present study, the expression patterns of nestin and vimentin in the spinal cords of rats with experimental autoimmune encephalomyelitis (EAE) and the response of cells containing filaments against acute autoimmune injury were examined by immunohistochemistry. Nestin immunostaining was only weakly detected in vascular endothelial cells but not in any cell types in the spinal cord in normal and adjuvant-immunized rats.

Expression of constitutive endothelial and inducible nitric oxide synthase in the sciatic nerve of Lewis rats with experimental autoimmune neuritis.

This study examined the expression of constitutive endothelial nitric oxide synthase (eNOS) and inducible NOS (iNOS) in the sciatic nerve of Lewis rats with experimental autoimmune neuritis (EAN). Western blot analysis showed that both eNOS and iNOS expressions in the sciatic nerves of rats increased significantly during the peak stage of EAN, but declined thereafter. Only minimal amounts of these enzymes were identified in normal rat sciatic nerves.