Local protein synthesis by BDNF is potentiated in hippocampal neurons exposed to ephrins

Miyata, Sumiko; Mori, Yuichiro; Fujiwara, Tatsuji; Ikenaka, Kazuhiro; Matsuzaki, Shinsuke; Oono, Kayoko; Katayama, Taiichi; Tohyama, Masaya

doi:10.1016/j.molbrainres.2004.10.034

Cited by 9 publications

(4 citation statements)

References 25 publications

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“…This currently include the use of fluorescent derivatives of cycloheximide to identify a specific site of protein synthesis (25) transfection of genetically engineered vectors for the expression of tagged mRNAs to visualize growing nascent peptide at sites of specific mRNA localization (26); expression of tagged translation initiation factors that serve as FRET pairs to specifically monitor translation initiation (24); and in situ spectral monitoring using the ratio between the peak areas of RNA and proteins as a measure of mRNA translation (28). The results presented in the current study demonstrate the ability of FtTM to quantitatively measure the process of translation and delineate the intracellular topography of protein synthesis activity under normal physiological conditions, in single cells and at subcellular resolution.…”

Section: Discussionmentioning

confidence: 99%

Quantitative single cell monitoring of protein synthesis at subcellular resolution using fluorescently labeled tRNA

Barhoom

Kaur

Cooperman

et al. 2011

Nucleic Acids Research

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We have developed a novel technique of using fluorescent tRNA for translation monitoring (FtTM). FtTM enables the identification and monitoring of active protein synthesis sites within live cells at submicron resolution through quantitative microscopy of transfected bulk uncharged tRNA, fluorescently labeled in the D-loop (fl-tRNA). The localization of fl-tRNA to active translation sites was confirmed through its co-localization with cellular factors and its dynamic alterations upon inhibition of protein synthesis. Moreover, fluorescence resonance energy transfer (FRET) signals, generated when fl-tRNAs, separately labeled as a FRET pair occupy adjacent sites on the ribosome, quantitatively reflect levels of protein synthesis in defined cellular regions. In addition, FRET signals enable detection of intra-populational variability in protein synthesis activity. We demonstrate that FtTM allows quantitative comparison of protein synthesis between different cell types, monitoring effects of antibiotics and stress agents, and characterization of changes in spatial compartmentalization of protein synthesis upon viral infection.

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Section: Discussionmentioning

confidence: 99%

Quantitative single cell monitoring of protein synthesis at subcellular resolution using fluorescently labeled tRNA

Barhoom

Kaur

Cooperman

et al. 2011

Nucleic Acids Research

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“…These include endoplasmic reticulum (ER) and Golgi membranes and also vesicle pools carrying either NMDA or AMPA receptor subunits (Kennedy and Ehlers 2006). The former, together with the presence of dendritic ribosomes and mRNAs (Steward and Schuman 2001;Sutton and Schuman 2005), raise the possibility that both cytosolic and integral membrane proteins can be locally synthesized, processed, and inserted into nascent and mature synapses, a concept supported by several recent publications (Ju et al 2004;Miyata et al 2005). However, several studies have provided evidence that prefabricated protein complexes contribute to the assembly of the PSD.…”

Section: Cellular Mechanisms Of Synapse Assemblymentioning

confidence: 99%

Synapse development: still looking for the forest, still lost in the trees

2006

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Synapse development in the vertebrate central nervous system is a highly orchestrated process occurring not only during early stages of brain development, but also (to a lesser extent) in the mature nervous system. During development, the formation of synapses is intimately linked to the differentiation of neuronal cells, the extension of their axons and dendrites, and the course wiring of the nervous system. Subsequently, the stabilization, elimination, and strengthening of synaptic contacts is coupled to the refinement of axonal and dendritic arbors, to the establishment of functionally meaningful connections, and probably also to the day-to-day acquisition, storage, and retrieval of memories, higher order thought processes, and behavioral patterns.

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“…In the developing brain, the final connectivity of a neuron is determined largely by signals received from the neurotrophin family of proteins, which are expressed in a laminar-specific pattern and influence the growth and complexity of the dendritic tree (McAllister et al, 1995 ). Of this family, brain-derived neurotrophic factor (BDNF) has a profound effect on pyramidal neurons in cortical layers 4 and 5, in addition to which it has demonstrated the capacity to target mRNAs to the synapse (Tongiorgi et al, 1997 ; Righi et al, 2000 ) and stimulate their local translation (Miyata et al, 2005 ). In particular, polysome profiling of active translation conducted by Schratt and colleagues demonstrated that BDNF is required to stimulate local translation of key synaptic components such as CamKII, NMDA receptors NR1, and NR3, PSD93 and LIMK-1 (Schratt et al, 2004 ).…”

Section: Neuronal Maturation With Bdnfmentioning

confidence: 99%

BDNF and the maturation of posttranscriptional regulatory networks in human SH-SY5Y neuroblast differentiation

Goldie

Barnett

Cairns

2014

Front. Cell. Neurosci.

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The SH-SY5Y culture system is a convenient neuronal model with the potential to elaborate human/primate-specific transcription networks and pathways related to human cognitive disorders. While this system allows for the exploration of specialized features in the human genome, there is still significant debate about how this model should be implemented, and its appropriateness for answering complex functional questions related to human neural architecture. In view of these questions we sought to characterize the posttranscriptional regulatory structure of the two-stage ATRA differentiation, BDNF maturation protocol proposed by Encinas et al. (2000) using integrative whole-genome gene and microRNA (miRNA) expression analysis. We report that ATRA-BDNF induced significant increases in expression of key synaptic genes, brain-specific miRNA and miRNA biogenesis machinery, and in AChE activity, compared with ATRA alone. Functional annotation clustering associated BDNF more significantly with neuronal terms, and with synaptic terms not found in ATRA-only clusters. While our results support use of SH-SY5Y as a neuronal model, we advocate considered selection of the differentiation agent/s relative to the system being modeled.

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Local protein synthesis by BDNF is potentiated in hippocampal neurons exposed to ephrins

Cited by 9 publications

References 25 publications

Quantitative single cell monitoring of protein synthesis at subcellular resolution using fluorescently labeled tRNA

Quantitative single cell monitoring of protein synthesis at subcellular resolution using fluorescently labeled tRNA

Synapse development: still looking for the forest, still lost in the trees

BDNF and the maturation of posttranscriptional regulatory networks in human SH-SY5Y neuroblast differentiation

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