Neuronal Nuclear Antigen (NeuN): A New Tool in the Diagnosis of Central Neurocytoma
Department of Pathology, Faculty of Medicine, Hacettepe University, Ankara, Turkey. Pathology - Research and Practice
(Impact Factor: 1.4).
02/2003; 199(7):463-8. DOI: 10.1078/0344-0338-00446
The use of neuronal nuclear antigen (NeuN) as a reliable neuronal marker in the differential diagnosis of clear cell neoplasms of the central nervous system was determined in a biopsy series of 23 cases. Immunohistochemical analyses were carried out by antisera against neuronal nuclear antigen, synaptophysin, neuron-specific enolase, microtubule-associated protein 2, and glial fibrillary acidic protein. All eight central neurocytomas were characteristically immunolabeled by NeuN. NeuN immunoreactivity was uniformly strong and basically located in the nuclei of neurocytes. Despite this uniform staining pattern of central neurocytomas, 12 cases of oligodendrogliomas and three cases of ependymoma were negative for NeuN. As the diagnostic criteria for central neurocytoma include immunohistochemical and/or ultrastructural evidence for neuronal differentiation, NeuN as a sensitive and specific neuronal marker in formalin-fixed, paraffin-embedded tissues may greatly facilitate the differential diagnosis of central neurocytomas.
Available from: Rebecca Jane Rylett
- "NeuN is a DNAbinding protein found exclusively in neurons with a molecular mass of 46–48 kDa (Lind et al., 2005; Mullen et al., 1992; Wolf et al., 1996). Although the function of NeuN is not known, it appears to be a nervous system-specific nuclear regulatory molecule (Soylemezoglu et al., 2003). This study illustrates some of the difficulties encountered when studying neuronal nuclear proteins by immunostaining, and provides solutions for exposing nuclear antigens to allow characterization of expression, distribution, and localization of endogenous nuclear proteins. "
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ABSTRACT: Immunohistochemical and immunofluorescence staining approaches are powerful tools for characterization of the endogenous protein expression and subcellular compartmentalization. However, several technical problems hamper identification of low-abundance nuclear proteins in archival formalin-fixed, paraffin-embedded human neural tissue. These include loss of protein antigenicity during tissue fixation and processing, and intrinsic auto-fluorescence associated with the tissue related to its fixation and the presence of lipofuscin. We evaluated several antigen retrieval methods to establish a strategy for detection of neuronal nuclear proteins in human spinal cord formalin-fixed, paraffin-embedded tissue. Thus, using immunostaining of the neuron-specific nuclear protein NeuN as the outcome measure, we found that heating tissue sections in an alkaline pH buffer unmasked protein epitopes most effectively. Moreover, staining by immunohistochemistry with diaminobenzidine tetrahydrochloride chromagen was superior to immunofluorescence labeling, likely due to the signal amplification steps included in the former approach. Auto-fluorescence in the tissue sections can be effectively reduced, but a sufficient fluorescence signal associated with specific antibody labeling could not be detected above this background for NeuN in the nucleus.
Journal of Neuroscience Methods 11/2005; 148(1):26-35. DOI:10.1016/j.jneumeth.2005.03.008 · 2.05 Impact Factor
Available from: Li-Hsien Lin
- "To examine the nature of nNOS - IR positive and / or VGLUT3 - IR positive cells we performed triple fluorescent immunostaining for VGLUT3 , nNOS and the neuronal marker , NeuN ( Magavi et al . , 2000 ; Pluchino et al . , 2003 ; Soylemezoglu et al . , 2003 ) . We incubated sections in a mixture of Guinea pig anti - VGLUT3 antibody ( 1 : 500 ) , sheep anti - nNOS antibody ( 1 : 1000 ) and mouse anti - NeuN ( 1 : 50 , Chemicon ) for 20 – 24 h at 25 8C . Sections were then incubated in a mixture of fluorescein - conjugated donkey anti"
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ABSTRACT: Earlier we reported that glutamate transporter (VGLUT) 2 and neuronal nitric oxide synthase (nNOS) are colocalized in some fibers and are present in apposing fibers in the nucleus tractus solitarii (NTS). Those findings provided anatomical support for a hypothesized physiological link between glutamate and nitric oxide (NO.) in the NTS. Recently a third class of VGLUT, VGLUT3, was identified, but its distribution in NTS and its anatomical relationship with nNOS have not been shown. In this study we tested the hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS. We perfused rats and obtained brain stem sections and nodose ganglion sections for immunofluorescent staining analyzed by confocal microscopy. The NTS contained moderate VGLUT3-immunoreactivity (IR), with the intermediate, medial and interstitial subnuclei containing higher VGLUT3-IR than other subnuclei. Although all three forms of VGLUT were present in the NTS, VGLUT3-IR was not colocalized with either VGLUT1-IR or VGLUT2-IR in either processes or cells in the brain stem. Cells and processes containing both VGLUT3-IR and nNOS-IR were noted in all NTS subnuclei and in the nodose ganglion. Triple immunofluorescent staining revealed that cells double-labeled for nNOS-IR and VGLUT3-IR were all additionally labeled for neuronal nuclear antigen (NeuN), a neuronal marker. These findings support our hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS.
Journal of Chemical Neuroanatomy 06/2005; 29(3):179-91. DOI:10.1016/j.jchemneu.2005.01.002 · 1.50 Impact Factor
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ABSTRACT: In this paper the applicability of the compact Green functions
technique in the problems of signal restoration and device
characterization is discussed. It is shown that the compact Green
functions, in a straightforward way, can be used to restore the incident
signal from the distorted signal which is received at the other end of
the line. Further, an exact method to reconstruct two of the electrical
parameters of a nonuniform transmission line from the transient response
is presented. This method may be used to characterize a microwave device
in terms of transmission line parameters
Signals, Systems, and Electronics, 1995. ISSSE '95, Proceedings., 1995 URSI International Symposium on; 11/1995
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