Young Chul Yang

Axotomy-induced dopaminergic neurodegeneration is accompanied with c-Jun phosphorylation and activation transcription factor 3 expression.

Accumulating evidence has shown that both phosphorylated c-Jun (pc-Jun) and activating transcription factor 3 (ATF3) were upregulated in a variety of tissue injuries and proposed to play an important role in cell death/survival. To elucidate the significance and functional role of these immediate-early genes during neuronal damage in the central nervous system, we examined temporal and spatial profiles of pc-Jun and ATF3 in dopaminergic neurons of the substantia nigra (SN) following transection of the medial forebrain bundle (MFB) in adult rats. Morphological characteristics of pc-Jun-positive dopaminergic neurons as well as microglial reaction in response to the axotomy-induced neurodegeneration were also investigated. Following MFB transection, both c-Jun phosphorylation and ATF3 were found in the nuclei of tyrosine hydroxylase-immunoreactive (TH-ir) neurons of the ipsilateral SN, but not in those of the contralateral SN. In the ipsilateral SN, the number of pc-Jun- and ATF3-positive nuclei was increased by 5-7 days post-lesion, and then progressively decreased probably due to the loss of neurons. Retrograde tracing with FluoroGold (FG) in hemi-axotomized rat brain demonstrated that none of the intact, unaxotomized (FG-ir) neurons was pc-Jun-positive, indicating phosphorylation of c-Jun occurs only in axotomized neurons. Concomitant co-localization of pc-Jun and ATF3 in the same TH-ir neuron was also demonstrated by triple immunofluorescence labeling. Many TH-ir neurons that underwent various steps of consecutive neurodegenerative changes retained pc-Jun in the condensed or fragmented nuclei. Moreover, numerous activated microglia, identified by both phagocytic (ED1) and MHC II (OX6) markers, closely apposed to these neurons throughout the entire neurodegenerative process, suggesting that they are actively phagocytosing dying neurons. Taken together, these results support the idea that pc-Jun and its putative dimeric partner ATF3 may be closely participating in axotomy-induced neurodegeneration.

Transplantation of human mesenchymal stem cells into the cisterna magna and its neuroprotective effects in a parkinsonian animal model

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases in the elderly, and therefore, the demand for effective therapies against PD has greatly increased. Therapeutic applications of stem cells have been considered to be one of the promising approaches in PD therapy. In the present study, the neuroprotective effects of hMSCs were evaluated in a parkinsonian rat model. The animal model was established by injecting 6-hydroxydopamine into the striatum of rats. Two weeks later, cultured hMSCs were transplanted into the cisterna magna. We subsequently identified changes in the expression of inflammatory cytokines, neurotrophic factors, microglial activation, and the survival rate of dopaminergic neurons in SNc. Behavioral recovery was also examined. The results indicated that hMSC transplantation increased the expression of anti-inflammatory cytokines as well as neurotrophic factors, and decreased the number of activated microglia. Compared to the sham-grafted group, relatively large numbers of TH-positive neurons were found in the ipsilateral SNc, and amphetamine-induced asymmetrical rotation was significantly reduced after hMSC transplantation. These findings suggest that hMSCs might be neuroprotective, probably through up-regulation of neurotrophic factors and anti-inflammatory cytokines, and this could have a functional impact in reversing PD symptoms.

Neurotoxin 6-aminonicotinamide affects levels of soluble proteins and enzyme activities in various tissues of golden hamsters.

The effects of neurotoxin 6-aminonicotinamide (6-AN) on the levels of soluble proteins and enzyme activities in various tissues of golden hamsters were investigated. SDS-polyacrylamide gel electrophoresis showed that a soluble spinal cord protein with molecular mass 75.0 kDa was present at a higher concentration in the treated group compared to that in the control while that of a molecular mass 64.8 kDa appeared to be missing. However, there were no noticeable differences in protein concentrations observed with the cerebrum, brain stem, and cerebellum. Similarly, treatment with 6-AN decreased the concentration of a soluble protein in pectoral muscle having molecular mass 97.2 kDa and increased those having molecular masses 207.4 and 32.1 kDa. In the kidney, soluble proteins with molecular masses 176.6 kDa was missing and those of molecular masses 97.6, 49, 43.3, and 33.8 kDa were decreased whereas those of molecular masses 64.7 and 33.1 kDa were increased. In the testis the soluble proteins with molecular masses 125.4, 88.7, 69.0, 31.2, 19.1, and 17.4 kDa were missing and those of molecular masses 97.0, 51.3, 42.0, 33.0, 27.2, and 22.6 kDa were present in lower amounts whereas those of molecular masses 311.5, 75.0, 64.0, 54.1, and 53.2 kDa were present in higher amounts. The specific activity of 6-phosphogluconate dehydrogenase was markedly increased in the liver but that of other tissues was not affected. Acetylcholinesterase activity was markedly reduced in the spleen but was enhanced in the intestine. Monoamine oxidase activity was markedly reduced in the brain stem, cerebrum, kidney, and liver. The results suggest that the changes in levels of soluble proteins and enzyme activities shown with golden hamster tissues by 6-AN administration were quite different from those shown with quail tissues.

Upregulated and prolonged differentiation potential of the ependymal cells lining the ventriculus terminalis in human fetuses

The ventriculus terminalis (VT) is a dilated cavity within the conus medullaris of the spinal cord. Although the VT was discovered in the mid-nineteenth century, little is known about its characteristics during development in human fetuses. Ependymal cells lining the cavities within the CNS retain high differentiation potential, and are believed to be responsible for the postnatal neurogenesis. To evaluate the differentiation capacity of the ependymal cells lining the VT during development, we examined glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) expression in the spinal cord of 18-24-week-old human fetuses. GFAP is a marker for the degree of ependymal cell differentiation in the human fetus, and PCNA is a well-known marker for cell division. Morphological characteristics of the VT were also examined. At the lower portion of the conus medullaris, the central canal abruptly expands dorsally to become the VT. Then the VT widens bilaterally while its anteroposterior diameter reduces gradually in a caudal direction. Finally, the VT becomes a narrow, transverse slit at the level of the lowermost conus medullaris. Compared with those lining the central canal, more numerous ependymal cells lining the VT showed more intensive GFAP and PCNA expression throughout all gestational ages examined. This suggests that, in the developing human spinal cord, ependymal cells lining the VT retain their differentiation potential, including a higher proliferative capacity, until a later stage of development than those lining the central canal.

CCL11 promotes angiogenic activity by activating the PI3K/Akt pathway in HUVECs

Abstract CCR3, the receptor for CCL11, is expressed on the surface of immune cells and even on non-immune cells. CCL11-CCR3 interactions can promote cell migration and proliferation. In this study, we investigated the effect of CCL11 on angiogenesis in HUVECs and also examined the molecular mechanisms of this process. We found that CCL11 induced mRNA transcription and protein expression of CCR3 in HUVECs. Moreover, the scratch wound healing assay and MTS proliferation assay both demonstrated that CCL11 promotes endothelial cell migration and induces weak proliferation. CCL11 directly induced microvessel sprouting from the rat aortic ring; these effects occurred earlier and to a greater extent than with VEGF stimulation. Furthermore, CCL11-induced phosphorylation of Akt was abolished by PI3K inhibitors. siRNA-mediated knockdown of CCR3 led to a significant reduction of PI3K phosphorylation. However, the phosphorylation levels of ERK1/2 were not changed, even after CCL11 treatment. Cumulatively, our data suggest that the CCL11–CCR3 interaction mainly activates PI3K/Akt signal transduction pathway in HUVECs.