Visual stimulation‐induced phosphorylation of neurofilament‐L in the visual cortex of dark‐reared rats

Hashimoto, Ryota; Nakamura, Yu; Imamura, Kazuyuki; Nakadate, Kazuhiko; Kashiwagi, Yujiro; Matsumoto, Nobuyoshi; Takeda, Masatoshi

doi:10.1046/j.0953-816x.2001.01747.x

Cited by 8 publications

(2 citation statements)

References 53 publications

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“…Cells were lysed with sodium dodecyl sulfate (SDS) buffer (50 mM Tris–HCl, pH 7.5, 0.5% SDS, 1 mM dithiothreitol). The lysates were sonicated at 30 amplitudes (Sonic&Materials, Danbury, CT, USA) in an ice‐bath four times, for 10 s each time, and then an aliquot of 50 μg protein was dissolved in the sample buffer for SDS–polyacrylamide gel electrophoresis and Western blotting as described previously [15]. Separated proteins were transferred onto a polyvinylidene difluoride membrane, which was incubated with a primary antibody against Src (monoclonal; Oncogene, San Diego, CA, USA), Fyn (polyclonal; Santa Cruz, Santa Cruz, CA, USA), c‐Yes (monoclonal; Wako Chemicals, Richmond, VA, USA), phospho‐Src (polyclonal against phospho‐Tyr416; Cell Signaling Technology, Beverly, MA, USA), postsynaptic density (PSD)‐95 (monoclonal; Upstate, Lake Placid, NY, USA) or SynGAP (polyclonal; Upstate).…”

Section: Methodsmentioning

confidence: 99%

Lithium‐induced inhibition of Src tyrosine kinase in rat cerebral cortical neurons: a role in neuroprotection against N‐methyl‐D‐aspartate receptor‐mediated excitotoxicity

et al. 2003

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The neuroprotective e¡ects of lithium, a mood stabilizer, against glutamate-induced excitotoxicity in rat cortical neurons were associated with a decrease in Tyr1472 phosphorylation of the N-methyl-D-aspartate (NMDA) receptor NR2B subunit and a loss of receptor activity. Since this receptor tyrosine phosphorylation is mediated by the Src-family tyrosine kinases, we investigated the e¡ects of lithium on the Src kinase activity. Levels of phosphorylated Src kinase at Tyr416, an index of Src activation, were reduced after treatment with LiCl (1 mM) for more than 3 days. Protein levels of Src-family kinases such as Src, Fyn, and Yes were unchanged by lithium treatment. The activities of cytosolic protein tyrosine kinase and protein phosphatase were also unchanged by lithium treatment, indicating the selectivity and the modulation. Moreover, the levels of postsynaptic densities (PSD) and SynGAP, the scaffolding proteins of the NMDA receptor complex, were unaltered by lithium. A Src kinase inhibitor, SU6656, and an NR2B antagonist, ifenprodil, partially blocked glutamate excitotoxicity. Our results suggest that lithium-induced inactivation of Src kinase contributes to this drug-induced NMDA receptor inhibition and neuroprotection against excitotoxicity. ß

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Section: Methodsmentioning

confidence: 99%

Lithium‐induced inhibition of Src tyrosine kinase in rat cerebral cortical neurons: a role in neuroprotection against N‐methyl‐D‐aspartate receptor‐mediated excitotoxicity

et al. 2003

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“…lation of NF-L (Hashimoto et al, 2001). Furthermore, in hippocampal slices of adult rats, the phosphorylation of NF-L by calcium/calmodulin-dependent protein kinase has occurred in a subpopulation of apical dendrites of pyramidal cells (Hashimoto et al, 2000).…”

Section: Lopez-picon Et Almentioning

confidence: 97%

Differential expression and localization of the phosphorylated and nonphosphorylated neurofilaments during the early postnatal development of rat hippocampus

López-Picón¹,

Uusi‐Oukari²,

Holopainen³

2003

Hippocampus

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Neurofilament (NF) proteins are expressed in most mature neurons in the central nervous system. Although they play a crucial role in neuronal growth, organization, shape, and plasticity, their expression pattern and cellular distribution in the developing hippocampus remain unknown. In the present study, we have used Western blotting and immunocytochemistry to study the low- (NF-L), medium- (NF-M), and high- (NF-H) molecular-weight NF proteins; phosphorylated epitopes of NF-M and NF-H; and a nonphosphorylated epitope of NF-H in the early postnatal (through P1-P21) development of the rat hippocampus. During the first postnatal week, NF-M was the most abundantly expressed NF, followed by NF-L, whereas the expression of NF-H was very low. Through P7-P14, the expression of NF-H increased dramatically and later began to plateau, as also occurred in the expression of NF-M and NF-L. At P1, no NF-M immunopositive cell bodies were detected, but cell processes in the CA1-CA3 fields were faintly immunopositive for NF-M and for the phosphorylated epitopes of NF-M and NF-H. At P7, CA3 pyramidal neurons were strongly immunopositive for NF-L and NF-H, but not for NF-M. The axons of granule cells, the mossy fibers (MFs), were NF-L and NF-M positive through P7-P21 but were NF-H immunonegative at all ages. Although they stained strongly for the phosphorylated NF-M and NF-H at P7, the staining intensity sharply decreased at P14 and remained so at P21. The cell bodies of CA1 pyramidal neurons and granule cells remained immunonegative against all five antibodies in all age groups. Our results show a different time course in the expression and differential cell type and cellular localization of the NF proteins in the developing hippocampus. These developmental changes could be of importance in determining the reactivity of hippocampal neurons in pathological conditions in the immature hippocampus.

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Neuroproteomics in stem cell differentiation

Beyer

Mix

Hoffrogge

et al. 2007

Proteomics Clinical Apps

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The term "proteome" is used to describe the entire complement of proteins in a given organism or in a system at a given time. Proteome analysis in neuroscience, also called "neuroproteomics" or "neuromics" is in its initial stage, and shows a deficit of studies in the context of brain development. It is the main objective of this review to illustrate the potential of neuroproteomics as a tool to unravel the differentiation of neural stem or progenitor cells to terminally differentiated neurons. Experimental results regarding the rat striatal progenitor model cell line ST14A are presented to illustrate the large rearrangements of the proteome during the differentiation process of neural progenitor cells and their modification by neurotrophic factors like the glial cell line-derived neurotrophic factor (GDNF). Thereby native stem cells and cells transfected with GDNF gene were investigated at the proliferative state and at seven time points up to 72 h after induction of differentiation. In addition, the immortalized human fetal midbrain stem cell line ReNcell VM was analyzed in order to detect stem cell differentiation associated changes of the protein profile. This review gives also an outlook on technical improvements and perspectives of application of neural stem cell proteomics.

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Visual stimulation‐induced phosphorylation of neurofilament‐L in the visual cortex of dark‐reared rats

Cited by 8 publications

References 53 publications

Lithium‐induced inhibition of Src tyrosine kinase in rat cerebral cortical neurons: a role in neuroprotection against N‐methyl‐D‐aspartate receptor‐mediated excitotoxicity

Lithium‐induced inhibition of Src tyrosine kinase in rat cerebral cortical neurons: a role in neuroprotection against N‐methyl‐D‐aspartate receptor‐mediated excitotoxicity

Differential expression and localization of the phosphorylated and nonphosphorylated neurofilaments during the early postnatal development of rat hippocampus

Neuroproteomics in stem cell differentiation

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