This study was performed to investigate the sequential changes of the retinal tissue in tissue culture condition. The human sensory retinal tissues were cultured for up to 2 weeks and 4 weeks, respectively. The initial changes showed the separation of the intercellular space and the consequent widening of the intercellular space with prolapse of cytoplasmic processes into the widened intercellular space. The internal limiting membrane was also separated from the inner retina, which led to the prolapse of the cytoplasm of the Muller cell. The growth of the Muller cell was most prominent during the 4-weeks' tissue culture period. These findings suggest that the Muller cell might contribute to the formation of cellular membrane in case of the defect of the internal limiting membrane in several pathologic conditions.
Adult ; Cell Membrane/ultrastructure ; Cells, Cultured ; Humans ; Male ; Middle Aged ; Neuroglia/*ultrastructure ; Retina/*ultrastructure

Development and differentiation of astrocytes and oligodendrocytes (OC) in the developing human fetal spinal cord (HFSC) have been investigated by the correlative analysis of light microscopic, EM, Golgi and immunocytochemical studies. The evidence is presented to suggest, (a) that radial glia are the first distinguishable neuroglial element among the cells within the ventricular zone, (b) that radial glia contains astrocyte-specific glial fibrillary acidic protein (GFAP), (c) that radial glia undergoes transformation into astroglial cells, (d) that "transitional forms" possessing the light and EM features of both astroglial and oligodendroglial cells appear just prior to the onset of myelination, and (e) the myelin-forming OC are most likely derived from radial glial cells, either directly or through intermediated astroglial forms.
Human ; In Vitro ; Microscopy, Electron ; Neuroglia/ultrastructure* ; Oligodendroglia/ultrastructure* ; Spinal Cord/embryology* ; Spinal Cord/ultrastructure

OBJECTIVETo understand the possible pathogenesis of sporadic multiple system atrophy (MSA).

METHODSThe immunoreactivity and ultrastructural features of glial cytoplasmic inclusions (GCIs) in 12 autopsy patients with MSA and 4 normal control groups were studied. All regional sections from each subject were evaluated with HE staining, Klüver-Barrera (KB), Holzer's, modified Gallyas-Braak's (GB) methods and immunohistochemical staining with alpha-synuclein and ubiquitin antibodies. Pontine white matter with abundant GCIs from case 1 was examined, using conventional electron microscopy, Gallyas-Braak's electron microscopy and immunoelectron microscopy.

RESULTSThe presence of GCIs as constantly demonstrated in all MSA patients. Strong alpha-synuclein immunoreactivity was observed in all of the ubiquitinated GCIs. However, the density of alpha-synuclein positive GCIs differed from case to case, and there was no relationship between the density of GCIs and age, sex, or MSA subtype. Ultrastructural features indicated that argyrophilic granule-associated filaments of about 25 nm in diameter were the predominant constituents of GCIs, and the anti alpha-synuclein antibody selectively labeled in these filaments. No GCIs and alpha-synuclein immunoreaction were found in control brain tissues.

CONCLUSIONSGCI was a pathognomonic change in sporadic MSA patients. Accumulation of alpha-synuclein in GCIs may occur during the early stags of MSA. Seletcive alpha-synuclein positive abnormal microtubules in GCIs therefore play an important role in the pathogenesis of MSA.

Aged ; Aged, 80 and over ; Female ; Humans ; Immunohistochemistry ; Inclusion Bodies ; ultrastructure ; Male ; Microscopy, Immunoelectron ; Middle Aged ; Multiple System Atrophy ; etiology ; metabolism ; pathology ; Nerve Tissue Proteins ; analysis ; Neuroglia ; ultrastructure ; Synucleins ; alpha-Synuclein

OBJECTIVETo study the effects of Ginkgo biloba extract 50 (GBE50) on inflammatory cytokines and glia cell injury in the prefrontal cortex and hippocampus of aging rats and its probable mechanism. Methods Totally 45 male SD rats were randomly divided into 4 groups, i.e., the normal control group (n=12), the model group (n=11), the low dose GBE50 group (n=10), and the high dose GBE50 group (n=12). The aging rat model was intraperitoneally injected with D-galactose to establish the aging model for 42 days. Starting from the 22nd day of modeling, rats in the low dose GBE50 group and the high dose GBE50 group were administered by gastrogavage with 75 mg/kg and 150 mg/kg respectively. The protein contents and mRNA expressions of IL-1beta, IL-6, and TNF-a in the prefrontal cortex and hippocampus of rats were detected by radioimmunoassay and Real-time fluorescence quantitative PCR assay respectively. The ultrastructural changes of glia cells in the hippocampal CA1 region were observed by transmission electron microscope. Results The protein contents and mRNA expressions of IL-1beta and TNF-alpha in the prefrontal cortex and the hippocampus of aging rats obviously increased when compared with the normal control group (P < 0.05, P < 0.01). The content of IL-6 in the hippocampus of aging rats obviously decreased (P < 0.01). Compared with the model group, the protein content and mRNA expression of IL-1beta in the prefrontal cortex and the hippocampus were obviously downregulated in the low and high dose GBE50 groups. The content of TNF-alpha in the prefrontal cortex was obviously downregulated in the low and high dose GBE50 groups, the content of TNF-alpha in the hippocampus was obviously downregulated in the low dose GBE50 group (P < 0.05, P < 0.01). The content of IL-6 in the prefrontal cortex of the low dose GBE50 group was up-regulated. The content of IL-6 in the hippocampus of the high dose GBE50 group was also upregulated. The mRNA expression of IL-6 in the prefrontal cortex of the low and high dose GBE50 groups obviously increased (P < 0.05, P < 0.01). Low and high dose GBE50 showed obvious recovery on the ultrastructural damage of glia cells in the hippocampal CA1 region.

CONCLUSIONSGBE50 showed inhibitive effects on the inflammatory reaction of nerves of aging rats. Its mechanism might be possibly correlated with its regulatory effects on the cytokines in the prefrontal cortex and the hippocampus, as well as the ultrastructures of glia cells in the prefrontal cortex and hippocampus to some degree.

Aging ; Animals ; Cytokines ; metabolism ; Ginkgo biloba ; Hippocampus ; cytology ; drug effects ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Male ; Neuroglia ; ultrastructure ; Plant Extracts ; pharmacology ; Prefrontal Cortex ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVECombine olfactory ensheathing glia (OEG) implantation with ex vivo non-viral vector-based neurotrophin-3 (NT-3) gene therapy in attempting to enhance regeneration after thoracic spinal cord injury (SCI).

METHODSPrimary OEG were transfected with cationic liposome-mediated recombinant plasmid pcDNA3.1(+)-NT3 and subsequently implanted into adult Wistar rats directly after the thoracic spinal cord (T9) contusion by the New York University impactor. The animals in 3 different groups received 4x10(5) OEG transfected with pcDNA3.1(+)-NT3 or pcDNA3.1(+) plasmids, or the OEGs without any plasmid transfection, respectively; the fourth group was untreated group, in which no OEG was implanted.

RESULTSNT-3 production was seen increased both ex vivo and in vivo in pcDNA3.1(+)-NT3 transfected OEGs. Three months after implantation of NT-3-transfected OEGs, behavioral analysis revealed that the hindlimb function of SCI rats was improved. All spinal cords were filled with regenerated neurofilament-positive axons. Retrograde tracing revealed enhanced regenerative axonal sprouting.

CONCLUSIONNon-viral vector-mediated genetic engineering of OEG was safe and more effective in producing NT-3 and promoting axonal outgrowth followed by enhancing SCI recovery in rats.

Animals ; Animals, Newborn ; Brain Tissue Transplantation ; methods ; Cells, Cultured ; DNA, Recombinant ; therapeutic use ; Disease Models, Animal ; Female ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Graft Survival ; genetics ; Growth Cones ; metabolism ; ultrastructure ; Nerve Regeneration ; genetics ; Neuroglia ; metabolism ; transplantation ; Neurotrophin 3 ; biosynthesis ; genetics ; Olfactory Bulb ; cytology ; transplantation ; Paralysis ; metabolism ; physiopathology ; therapy ; Plasmids ; genetics ; Rats ; Rats, Wistar ; Recovery of Function ; genetics ; Spinal Cord Injuries ; metabolism ; physiopathology ; therapy ; Treatment Outcome ; Up-Regulation ; genetics