Histological and lectin histochemical studies in the vomeronasal organ of the Korean black goat, Capra hircus coreanae.

We examined the localization of olfactory marker protein (OMP), protein gene product9.5 (PGP9.5), and glycan diversity in the vomeronasal organ (VNO) of the Korean black goat (Capra hircus coreanae) during the prenatal and postnatal periods using immunohistochemistry and lectin histochemistry. In fetal and 1-day-old goats, OMP was occasionally identified in receptor cells of the VNO sensory epithelium, and PGP9.5 was localized in both the sensory and non-sensory epithelia. In VNO from adult goats, OMP was abundant in the sensory epithelium and scarce in single cells of the non-sensory epithelium. These results suggest that OMP production is initiated in the VNO sensory epithelium (VNE) during the fetal stage, and that its activity is increased in adult VNO receptor cells and solitary cells in the non-sensory epithelium (VNSE). Furthermore, the free borders of the sensory epithelia were positive for 7 lectins, and 6 lectins were moderately and/or highly abundant in receptor cells. Supporting and basal cells, and nerve bundles had similar expression patterns. In VNE, 7 lectins were observed in the free border, and 6 in ciliated, goblet, and basal cells, and in gland acini. The intensities of WGA, LCA, and PNA were high in VSE receptor cells, and the intensity of PNA was high in ciliated cells of the VNSE. The other 3 lectins showed similar patterns throughout development. Collectively, these results confirm that the Korean black goat VNO starts developing during the late fetal stages and differentiates further after birth.

Hesperidin improves motor disability in rat spinal cord injury through anti-inflammatory and antioxidant mechanism via Nrf-2/HO-1 pathway.

Spinal cord injury (SCI) is associated with inflammation with concurrent oxidative stress and glial activation. The aim of this study was to evaluate whether hesperidin, a representative flavonoid in citrus fruits, ameliorates SCI-induced motor dysfunction and neuro-pathologic degeneration in rat model. Rats received hesperidin (100 mg/kg body weight/daily, oral administration) from 7 days prior to SCI to 7 days post SCI. Behavioral test was done on rats with SCI until 6 weeks. For the study of inflammatory molecules in SCI rats with hesperidin treatment, rats were sacrificed at day 4 post SCI, and spinal cords were collected and studied histopathologically. Behavioral tests on hind-limbs of rats with SCI revealed that treatment of hesperidin in rats with SCI significantly ameliorate the hind-limb paralysis beginning at day 21 post SCI. Hesperidin treatment in rats with SCI reduced the neuropathological changes (e.g., hemorrhage, inflammatory cell infiltration, and tissue loss) and pro-inflammatory cytokines including tumor necrotic factor-α and interleukin-1ß. In addition, oxidative stress related molecules including superoxide dismutase, catalase, nuclear factor erythroid 2-related factor-2 and heme oxygenase-1 were also increased by hesperidin treatment. Furthermore, Fe2+, bilirubin and p38 mitogen activated protein kinase, these by-product of heme catabolism in serum and spinal cord of rats with hesperidin-treatment groups were significantly increased compared with those of vehicle-treatment group. Collectively, this study implies that hesperidin accelerates recovery of locomotor function and tissue repair of damaged spinal cord, with concurrent upregulation of heme oxygenase-1 as far as rat SCI model is concerned.

Unilaminar follicular cells transiently express galectin-3 during ovarian folliculogenesis in pigs.

The localization of galectin-3, a ß-galactoside-binding animal lectin, was immunohistochemically studied in the ovaries of pigs to determine its expression in ovarian folliculogenesis. Various stages of ovarian follicles were identified in the ovaries of adult pigs. Galectin-3 was immunostained in the squamous follicular cells surrounding oocytes in primordial follicles and in the unilaminar granulosa cells of primary follicles, but not in oocytes of multilaminar follicles (including primary, secondary, and tertiary Graafian follicles). As in adult ovaries, galectin-3 immunoreactivity was prominent in the unilaminar follicles in neonatal ovaries. Galectin-3 was also immunolocalized in the luteal cells in the corpus luteum and granulosa cells of atretic follicles as well as in interstitial macrophages in porcine ovaries. Collectively, these results suggest that galectin-3 is transiently expressed in follicular cells in the unilaminar ovarian follicles (primordial and primary) but not in multilaminar ovarian follicles (primary to tertiary), implying that galectin-3 is embryologically involved in ovum generation.

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

Expression of osteopontin (OPN) was investigated in the spinal cords of rats with clip compression injury. Western blot analysis demonstrated that OPN protein increased significantly in the spinal cord during the early stages after injury. The increased expression of OPN was partially paralleled by that of proliferating cell nuclear antigen (PCNA). Immunohistochemical staining showed that OPN was expressed in proliferating activated microglia/macrophages in core lesions and in some astrocytes at the periphery of lesions. These results indicate that expression of OPN protein increases mainly in activated microglia/macrophages after spinal cord injury, suggesting that OPN is related to cell proliferation during the early stages after injury, probably leading to tissue remodeling.

Upregulation of CD44 expression in the spinal cords of rats with clip compression injury.

The expression of the extracellular matrix phosphoglycoprotein CD44 after compression injury of the spinal cord was examined in rats. Western blot analysis of tissues harvested on days 0 (sham), 1, 4 and 7 post-injury showed significant increases in CD44 expression from 1 to 7 days after compression injury compared to sham-operated controls. Immunohistochemistry revealed that CD44 was constitutively expressed in some astrocytes in sham-operated controls. At days 4-7 post-injury, CD44 was intensely expressed in astrocytes in the periphery of lesions, and in myelin sheaths, vessels, and the majority of inflammatory cells including macrophages in core lesions. The finding that expression of CD44 was upregulated after spinal cord injury suggests that CD44 contributes to cell adhesion and glial cell attraction during the early stages after spinal cord injury, and may thus promote remodeling of injured spinal cords.

Sodium salicylate-induced amelioration of experimental autoimmune encephalomyelitis in Lewis rats is associated with the suppression of inducible nitric oxide synthase and cyclooxygenases.

We studied the effects of oral administration of sodium salicylate on the expression of the pro-inflammatory mediators, nitric oxide synthase (iNOS) and cyclooxygenase- (COX-) 1 and 2, in rats with experimental autoimmune encephalomyelitis (EAE). Sodium salicylate (200 mg/kg) was administered orally for 13 days after the induction of EAE by immunization with guinea pig myelin basic protein and complete Freund's adjuvant. The onset (P<0.0001) and severity (P<0.05) of EAE paralysis in salicylate-treated animals were delayed and suppressed significantly compared with vehicle-treated controls. Western blot analysis showed that expression of COX-2 and iNOS, but not COX-1, decreased significantly in the spinal cords of salicylate-treated rats compared with vehicle-treated controls (P<0.05) and this finding was paralleled by immunohistochemical observations. These results suggest that the amelioration by salicylate of paralysis in rats with EAE is mediated in part by the suppression of COX and iNOS.

Activation of embryonic intermediate filaments contributes to glial scar formation after spinal cord injury in rats.

The expression of two intermediate filaments, nestin and vimentin, was studied in spinal cord injury (SCI) to elucidate their roles in the formation of glial scars. Rats were sacrificed 1, 4, and 7 days after induction of compression injury of the spinal cord using an aneurysm clip. The affected spinal cords were studied using antibodies against nestin and vimentin intermediate filaments. One day after spinal cord injury, some clusters of nestin-positive vessels were detected in the center of the injury, but few were seen in other cell types. Vimentin immunostaining was detected in some glial cells in the center and its level of immunoreactivity was enhanced in the ependymal cells of the central canal. On days 4 and 7 after spinal cord injury, astrocytes and some ependymal cells in the central canal were stained positively for nestin and increased expression of nestin was observed in vessels. Vimentin was detected in some macrophages and astrocytes in the lesions. Nestin was co-localized with glial fibrillary acidic protein in some glial cells in SCI. These findings imply that spinal cord cells in adult animals have embryonic capacity, and these cells are activated after injury, which in turn contributes to repair of spinal cord injury through formation of a glial scar.

Activation of mitogen-activated protein kinases in experimental autoimmune encephalomyelitis.

The expression of mitogen-activated protein (MAP) kinases, including extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal protein kinase (JNK), and p38, was analyzed in experimental autoimmune encephalomyelitis (EAE) in rats. Western blot analysis showed that the three MAP kinases (phosphorylated ERK (p-ERK), p-JNK, and p-p38) were increased significantly in the spinal cords of rats with EAE at the peak stage as compared with the levels in controls (p<0.05), and both p-ERK and p-JNK declined slightly in the recovery stage of EAE. Immunohistochemistry showed that p-ERK was constitutively expressed in brain cells, including astroglial cells, and showed enhanced immunoreactivity in those cells in EAE, while some T cells and macrophages were weakly immunopositive for p-ERK in EAE lesions. Both p-JNK and p-p38 were intensely immunostained in T cells in EAE lesions, while a few glial cells and astrocytes were weakly positive for both. Taking all these facts into consideration, we postulate that increased expression of the phosphorylated form of each MAP kinase plays an important role in the initiation of acute monophasic EAE. Differential expression of three MAP kinases was discerned in an animal model of human autoimmune central nervous system diseases, including multiple sclerosis.