Cortical dysplasia is a cause of intractable epilepsy and a candidate for surgical resection to control epileptic attacks. The neuronal cytomegaly and balloon cell change are the diagnostic hallmarks of cortical dysplasia. Little research has been performed about the normal-sized dysplastic neuron which has complex arborizing dendrites and lacks in its polarity. The aim of this study was to define the histopathologic characteristics of the neurons in cortical dysplasia. Twelve cases of cortical dysplasia who underwent partial lobectomy for intractable seizures were selected and immunohistochemical staining for NF-M/H, MAP2, tau, and ubiquitin was performed. The perikarya and dendrite of dysplastic neurons were more intensely labeled with antibodies for the high and medium molecular weight neurofilament proteins (NF-M/H) than normal neurons. Immunoreactivity with the MAP2 antibody expressed mainly within the somatodendritic regions was present in the dysplastic or normal neurons without any significant difference in intensity. The complex arborizing dendrites of dysplastic neurons were easily identified due to pronounced immunoreactivity within the somatodendritic regions. Immunoreactivity with the primary antibody against tau and ubiquitin was present in the normal-looking neurons as well as the dysplastic neurons. This study suggests that the dysplastic neurons in cortical dysplasia are accompanied by changes of cytoskeletal neurofilaments, and the immunohistochemical stains for NF-M/H, MAP2, tau, and ubiquigin are useful to detect them.
Antibodies ; Coloring Agents ; Dendrites ; Epilepsy ; Malformations of Cortical Development* ; Molecular Weight ; Neurofilament Proteins ; Neurons ; Seizures ; Ubiquitin

OBJECTIVETo identify the properties of nerve fibers of dogs by immunohistochemical staining method.

METHODSIntact bone blocks above the inferior alveolar nerve canal were cut from the medial of the second premolar to the distal of the third premolar of healthy adult Beagle dogs of 18 months, embedded to make hard tissue sections, stained with S100 and neurofilament protein (NFP) antibodies, and finally observed the nerve distribution under the microscope.

RESULTSThe distribution of S100 positive tissue in the periodontal ligament of dogs showed the following patterns: bundles of densely gathered rings with different diameters, filaments accompanied by lumens, free endings and deep-dyeing oval lamellasome. The location of NFP positive tissue was similar to that of S100 positive tissue, but the distribution of these NFP positive filaments with various diameters showed largely as bundles, free ending and branches scattering in periodontal membrane.

CONCLUSIONSWe may firstly distinguish the structure of the nerve fibers in periodontal ligament of nerve distribution, and then judge the categories of the nerve fibers by S100 immunohistochemistry furtherly according to comparison of the thickness of neural axon by NFP immunohistochemistry, and finally distinct the function and attribute of the nerve fibers in the periodontal ligament of dogs. 
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Animals ; Bicuspid ; Cytoskeleton ; Dogs ; Immunohistochemistry ; Mandibular Nerve ; Nerve Fibers ; Nervous System ; Neurofilament Proteins ; Periodontal Ligament

BACKGROUND: Neuronal migration disorder (NMD) is one of the causes of medically intractable epilepsy. As neurosurgical treatments for medically intractable epilepsy have expanded recently, precise histopathologic diagnosis is required. Histopathologic grading of NMD is important due to its association with neocortical development and expectation of prognosis. Many studies revealed abnormalities of neuronal cytoskeletal protein in abnormal neuronal cells of NMD. METHODS: We performed immunohistochemical staining for neurofilament protein (NF) subtypes, one of the neuronal cytoskeletal proteins, and investigated the staining pattern of specific cells in each grade of NMD. RESULTS: NF-L was more intensely labeled in perikarya, dendrites, and axons of normal or small sized dysplastic neurons, cytomegalic neurons, and balloon cells than of normal-looking neurons. Furthermore, positive reaction was more intense in high-grade lesion. NF-H and NF-M were mainly positive in the axons of gray and white matter and weakly positive in a few cytomegalic neurons and some balloon cells. CONCLUSION: NF-L is a better marker than NF-H and NF-M for the detection of normal or small sized dysplastic neurons, cytomegalic neurons, and balloon cells and for grading of NMD.
Axons ; Cerebral Cortex ; Cytoskeletal Proteins ; Dendrites ; Diagnosis ; Epilepsy ; Nervous System Malformations ; Neurofilament Proteins ; Neuronal Migration Disorders* ; Neurons* ; Prognosis

Phosphatidylinositol phosphates (PtdInsPs) are ubiquitous membrane phospholipids that play diverse roles in cell growth and differentiation. To clarify the regulation mechanism acting on neurofilament light chain (NF-L) self assembly, we examined the effects of various PtdInsPs on this process. We found that PtdInsPs, including PI(4,5)P2, directly bind to the positively charged Arg54 of murine NF-L, and this binding promotes NF-L self assembly in vitro. Mutant NF-L (R53A/R54A) proteins lacking binding affinity to PtdInsPs did not have the same effect, but the mutant NF-L proteins showed greater self assembly than the wild-type in the absence of any PtdInsP. These results collectively suggest that Arg54 plays a pivotal role in NF-L self assembly by binding with PtdInsPs.
Animals ; Fluorescent Antibody Technique ; Mice ; Mutation/genetics ; Neurofilament Proteins/genetics/*metabolism ; Phosphatidylinositol Phosphates/*metabolism ; Phospholipase C gamma/metabolism ; *Protein Multimerization

Retinal horizontal cell is second-order neuron that integrates the information from photoreceptors over large retinal areas, mediating the lateral spread of visual signals to distant retina. The Neurofilament proteins, considered as neuronal markers, have been imunolocalized to mammalian retinal horizontal cess. However, the immunolabeling of Neurofilament in human, has been focused on the studies of visual pathway and large ganglion cells The goal of this study is to see whether human retinal horizontal cells are indeed neuronal nature or glial nature by immunogold labeling for electron microscopy. The sections of 65 year-old human retina showed the expression of Neurofilament by horizontal cells, which confirms the evidence of human retinal horizontal cell as neuronal nature.
Aged ; Ganglion Cysts ; Humans* ; Microscopy, Electron ; Negotiating ; Neurofilament Proteins ; Neurons ; Retina ; Retinal Horizontal Cells* ; Retinaldehyde* ; Visual Pathways

OBJECTIVETo screen the proteins with differential expression levels in the cerebral tissue of hens exposed to tri-ortho-cresyl phosphate (TOCP), and to provide target proteins for studying the mechanism of organophosphoms ester-induced delayed neurotoxicity (OPIDN).

METHODSThirty two adult Roman hens were randomly divided into four groups: TOCP group was exposed to 1000 mg/kg TOCP, PMSF group was exposed to 40 mg/kg PMSF, PMSF plus TOCP group was exposed to 40 mg/kg PMSF and after 24 h exposed to 1000 mg/kg TOCP, control group was exposed to normal saline. All hens exposed to chemicals by gastro-intestine for 5 days were sacrificed, and the cerebral tissue were dissected and homogenized in ice bath. Total proteins extracted from the cerebral tissue were separated by isoelectric focusing as the first dimension and SDS-PAGE as the second dimension. The 2-DE maps were visualized after silver staining and analyzed by Image Master 2D software. At last ,the expressed protein spots were identified by Mass spectrometry.

RESULTSFrom total proteins in TOCP group, the PMSF plus TOCP group and PMSF group, 1185, 1294 and 1063 spots were detected, respectively. One thousand three hundred thirty two spots from total proteins in control group were detected. The match rates of protein spots in TOCP group, the PMSF plus TOCP group and PMSF group were 78.32 %, 79.56 % and 80.93%, respectively. There were 235 protein spots with differential expression levels between TOCP group and control group, which included 158 up regulation spots and 77 down regulation spots. According to the PMSF features, there were 102 spots with differential expression levels between TOCP group and control group and without differential expression levels between TOCP group and PMSF plus TOCP group, among them there were 13 spots with 4 fold differential expression levels between TOCP group and control group and without differential expression levels between TOCP group and PMSF group. Seven protein spots (homer-1b, Destrin, heat shock protein 70, eukaryotic translation initiation factors, proteasome alpha1 subunit, lactate dehydrogenase B, glutamine synthetase) were detected by Mass spectrometry.

CONCLUSIONThere are 112 protein spots with differential expression levels of the cerebral tissue in TOCP group, which may be related to OPIDN, among them 13 protein spots with differential expression levels are associated closely with OPIDN. Seven protein spots detected by Mass spectrometry may be related to the mechanism induced by OPIDN.

Animals ; Brain ; metabolism ; Cerebrum ; drug effects ; metabolism ; Chickens ; Neurofilament Proteins ; metabolism ; Phenylmethylsulfonyl Fluoride ; toxicity ; Proteome ; analysis ; Tritolyl Phosphates ; toxicity

OBJECTIVETo investigate the dynamic alterations of neurofilament subunits (NF) in sciatic nerve of hens with organophosphorus ester induced the delayed neurotoxicity or neuropathy (OPIDN).

METHODSHens with OPIDN were produced by giving 30 mg/kg methamidophos subcutaneously to the 10-month-old Roman hens daily for 15 days, and sacrificed after manifesting neurotoxic clinical signs on the 2nd, 10th, and 23rd day respectively. The sciatic nerves were dissected, homogenized and centrifuged. The levels of NF in supernatant and pellet of sciatic nerves were examined by Western blotting respectively at different time from 2 to 23 days.

RESULTSIntegrated optional density (IOD) of high molecular weight neurofilament (NF-H) in sciatic nerve pellet of hens on the day 2, 10, 23 after appearance of OPIDN were 145,117 +/- 17,038, 55,917 +/- 17,333 and 45,038 +/- 6,662 respectively. As compared with the control group (78,875 +/- 22,569), the contents of NF-H in pellet were increased by 84% on day 2, and decreased by 29% and 43% on day 10 and 23 respectively. IOD of NF-H in supernatant of sciatic nerves were 4,709 +/- 1,739, 12,337 +/- 3,205 and 16,745 +/- 931, which were reduced significantly as compared with the control (44,083 +/- 6,895) at three different times. There was no significant difference in IOD of middle molecular weight neurofilament (NF-M) between control group (27,925 +/- 2,660) and on day 2 (31,493 +/- 4,625) in pellet. Those were 19,367 +/- 2,746 and 6,612 +/- 1,119 respectively on day 10 and day 23 in pellet of hen's sciatic nerve, which were much less than that in control. Little were detected in supernatant on day 10, and the IOD of NF-M were 3,196 +/- 269 and 5,206 +/- 1,292 on day 2 and day 23 respectively, which were lessened by 81% and 70% as compared with the control (17,243 +/- 3,232). In sciatic nerve pellet of hens, IOD of low molecular weight neurofilament (NF-L) on day 2 was 39,211 +/- 3,800, which was much higher than that in the control (28,749 +/- 9,319). There were no significant differences between IOD on day 10 (27,974 +/- 3,611), day 23 (21,507 +/- 2,286) and the control. There was no detection both on day 2 and 10 in supernatant of sciatic nerve, and IOD of NF-L were 5,962 +/- 1,929 on day 23, which were reduced significantly compared with the control (11,897 +/- 352).

CONCLUSIONThe alterations of NF in sciatic nerve might contribute to the occurrence and development of OPIDN.

Animals ; Chickens ; Female ; Insecticides ; toxicity ; Neurofilament Proteins ; metabolism ; Organothiophosphorus Compounds ; toxicity ; Sciatic Nerve ; drug effects ; metabolism ; pathology ; Toxicity Tests

The long belief that dental primary afferent (DPA) neurons are entirely composed of nociceptive neurons has been challenged by several anatomical and functional investigations. In order to characterize non-nociceptivepopulation among DPA neurons, retrograde transport fluorescent dye was placed in upper molars of rats and immunohistochemical detection of peripherin and neurofilament 200 in the labeled trigeminal ganglia was performed. As the results, majority ofDPA neurons were peripherin-expressing small-sized neurons, showing characteristic ofnociceptive C-fibers. However, 25.7% of DPA were stained with antibody against neurofilament 200, indicating significant portion of DPA neurons are related to large myelinated Abeta fibers. There were a small number of neurons thatexpressed both peripherin and neurofilament 200, suggestive of Adelta fibers. The possible transition of neurochemical properties by neuronal injury induced by retrograde labeling technique was ruled out by detection of minimal expression of neuronal injury marker, ATF-3. These results suggest that in addition to the large population of C-fiber-related nociceptive neurons, a subset of DPA neurons is myelinated large neurons, which is related to low-threshold mechanosensitive Abeta fibers. We suggest that these Abeta fiber-related neurons might play a role as mechanotransducers of fluid movement within dentinal tubules.
Animals ; Dentin ; Intermediate Filament Proteins ; Membrane Glycoproteins ; Molar ; Myelin Sheath ; Nerve Tissue Proteins ; Neurofilament Proteins ; Neurons ; Neurons, Afferent ; Nociceptors ; Rats ; Trigeminal Ganglion

BACKGROUNDAdipose tissue-derived stromal cells (ADSCs) can be greatly expanded in vitro, and induced to differentiate into multiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic, and adipogenic cells. This study was designed to investigate the possibility of ADSCs differentiating into neurons.

METHODSAdipose tissue from rats was digested with collagenase, and adherent stromal cells were cultured. A medium containing a low concentration of fetal bovine serum was adopted to induce the cells to differentiate. ADSCs were identified by immunocytochemistry, and semi-quantitative RT-PCR was applied to detect mRNA expression of neurofilament 1 (NF1), nestin, and neuron-specific enolase (NSE).

RESULTSNestin-positive cells were found occasionally among ADSCs. ADSCs were found to express NSE mRNA and nestin mRNA, but not NF1 mRNA. ADSCs could differentiate into neuron-like cells in a medium composed of a low concentration of fetal bovine serum, and these differentiated cells displayed complicated neuron-like morphologies.

CONCLUSIONSThe data support the hypothesis that adipose tissue contains stem cells capable of differentiating into neurons. These stem cells can overcome their mesenchymal commitment, and may represent an alternative autologous stem cell source for CNS cell transplantation.

Adipose Tissue ; cytology ; Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Immunohistochemistry ; Intermediate Filament Proteins ; analysis ; Nerve Tissue Proteins ; Nestin ; Neurofilament Proteins ; analysis ; Neurons ; cytology ; Phenotype ; Phosphopyruvate Hydratase ; analysis ; Rats

Composite pheochromocytoma or paraganglioma of the adrenal gland is a well-recognized, yet extremely rare tumor with only one case reported in Korea. We report a case of incidentally found composite pheochromocytoma and ganglioneuroma of the adrenal gland in a 44-year-old female composed of intermingled components of pheochromocytom, ganglioneuroma, and cells with intermediate features. On immunohistochemical staining, the pheochromocytoma component was positive for synaptophysin and chromogranin, but negative for S-100 protein. Staining for the S-100 protein revealed sustentacular cells which formed a peripheral coat around the "Zellballen" and Schwann cells. The Fontana-Masson stain defined neuromelanin granules of ganglion cells and the ganglion cells expressed neural markers such as neurofilament proteins. Ultrastructural findings revealed pheochromocytes with a round or ovoid nucleus and occasionally prominent nucleolus containing numerous adrenaline and noradrenaline granules.
Adrenal Glands ; Adult ; Electrons ; Epinephrine ; Female ; Ganglion Cysts ; Ganglioneuroma ; Humans ; Korea ; Melanins ; Neurofilament Proteins ; Norepinephrine ; Paraganglioma ; Pheochromocytoma ; S100 Proteins ; Schwann Cells ; Silver Nitrate ; Synaptophysin