Chapter 11 Mechanisms of neuronal plasticity as analyzed at the single cell level

Eberwine, James; Cao, Yanxiang; Nair, Suresh; Miyashiro, Kevin; Mackler, Scott A.; Finnell, Richard H.; Surmeier, James D.; Dichter, Marc A.

doi:10.1016/s0079-6123(08)63289-6

Cited by 11 publications

(7 citation statements)

References 17 publications

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“…In spinal cord, the most intense staining is in the large motor neurons of the anterior horn and in the large neurons of the intermediate zone . Single cell RT-PCR [Eberwine et al, 1995] a sophisticated technique that analyzes the message repertoire of an individual cell, will deter- mine whether MAP-2a coexists with MAP-2b in the same cell or whether there are mature neurons expressing only MAP-2a. Transient transfection of MAP-2a in COS cells resulted in expression of full-length MAP-2a protein, which co-migrated with the MAP-2a expressed in rat brain and was also recognized by a number of MAP-2 antibodies that spanned the entire MAP-2 molecule.…”

Section: Discussionmentioning

confidence: 99%

Molecular and functional characteristics of MAP-2a: Ability of MAP-2a versus MAP-2b to induce stable microtubules in COS cells

Kalcheva

Rockwood

Kress

et al. 1998

Cell Motil. Cytoskeleton

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Section: Discussionmentioning

confidence: 99%

Molecular and functional characteristics of MAP-2a: Ability of MAP-2a versus MAP-2b to induce stable microtubules in COS cells

Kalcheva

Rockwood

Kress

et al. 1998

Cell Motil. Cytoskeleton

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“…cDNA synthesis, amplified R NA amplification, and e xpression profiles f rom single isolated cells. During whole-cell patch-clamp recording, the electrode contained avian myeloblastoma virus (AM V) reverse transcriptase and oligo-dT 24 -T7 primer (Eberwine et al, 1995). C ellular contents were aspirated into the patch pipette with a small amount of suction after a 10 min recording.…”

Section: Methodsmentioning

confidence: 99%

“…cDNA synthesis was completed in vitro at 37°C for 60 min. The cDNA was resuspended in 20 l of H 2 O, heated at 95°C for 3 min, and converted into double-stranded template as described previously (Eberwine et al, 1995). T wenty percent of the cDNA template was used to synthesize amplified RNA (aRNA) in 40 mM Tris, pH 7.4, 10 mM NaC l, 10 mM MgC l 2 , 0.01 mM dithiothreitol, 500 M ATP, GTP, C TP, and UTP, 20 U of RNAsin, and 100 U of T7 RNA polymerase at 37°C for 3-4 hr.…”

Section: Methodsmentioning

confidence: 99%

“…T wenty percent of the cDNA template was used to synthesize amplified RNA (aRNA) in 40 mM Tris, pH 7.4, 10 mM NaC l, 10 mM MgC l 2 , 0.01 mM dithiothreitol, 500 M ATP, GTP, C TP, and UTP, 20 U of RNAsin, and 100 U of T7 RNA polymerase at 37°C for 3-4 hr. aRNA was synthesized into doublestranded cDNA and used as a template for a second round of amplification as described previously (Eberwine et al, 1995). Ten nanograms of random hexamers with AM V reverse transcriptase were used to prime the first-strand cDNA synthesis.…”

Section: Methodsmentioning

confidence: 99%

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Rab3A Is Required for Brain-Derived Neurotrophic Factor-Induced Synaptic Plasticity: Transcriptional Analysis at the Population and Single-Cell Levels

et al. 2001

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Brain-derived neurotrophic factor (BDNF) modulates synaptic strength in hippocampal neurons, in addition to promoting survival and differentiation. To identify genes involved in trophic regulation of synaptic plasticity, we have used a multidisciplinary approach of differential display and family-specific slot blots in combination with whole-cell patch-clamp recordings of dissociated hippocampal neurons. Three hour exposure to BDNF elicited a 2.6-fold increase in synaptic charge and a concomitant induction of 11 genes as revealed by differential display, including the small GTP-binding vesicular trafficking protein Rab3A and the enzyme guanylate cyclase (GC). Slot blot analysis on a population of neurons confirmed an average of 3.1-fold induction of these clones. In contrast, individual pyramidal-like neurons that were first characterized electrophysiologically in the presence of BDNF and subjected to transcriptional analysis displayed more robust increases (4.8-fold), emphasizing the neuronal heterogeneity. Transcriptional changes of Rab3A and GC were accompanied by translational regulation, shown by Western blot analysis. Furthermore, a number of GC-associated and Rab3A effector molecules were induced by BDNF at either the gene or protein levels. The functional role of Rab3A in BDNF-induced synaptic plasticity was assessed using cells derived from Rab3A knock-out mice. These neurons failed to show an increase in synaptic charge in response to BDNF at 10 min; however a late response to BDNF was detected at 20 min. This late response was similar in time course to that induced by postsynaptic activation of glutamate receptors. Our results demonstrate a requirement for Rab3A and may reveal a temporal distinction between presynaptic and postsynaptic mechanisms of BDNF-induced synaptic plasticity associated with learning and memory.

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“…Neostriatal neurons from 3-to 4-week-old mice were acutely dissociated by using procedures similar to those described (25). …”

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confidence: 99%

Regulation of cyclin-dependent kinase 5 and casein kinase 1 by metabotropic glutamate receptors

Liu

Hong

Ule

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

120

129

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Cyclin-dependent kinase 5 (Cdk5) is a multifunctional neuronal protein kinase that is required for neurite outgrowth and cortical lamination and that plays an important role in dopaminergic signaling in the neostriatum through phosphorylation of Thr-75 of DARPP-32 (dopamine and cAMP-regulated phosphoprotein, molecular mass 32 kDa). Casein kinase 1 (CK1) has been implicated in a variety of cellular functions such as DNA repair, circadian rhythm, and intracellular trafficking. In the neostriatum, CK1 has been found to phosphorylate Ser-137 of DARPP-32. However, first messengers for the regulation of Cdk5 or CK1 have remained unknown. Here we report that both Cdk5 and CK1 are regulated by metabotropic glutamate receptors (mGluRs) in neostriatal neurons. (S)-3,5-dihydroxyphenylglycine (DHPG), an agonist for group I mGluRs, increased Cdk5 and CK1 activities in neostriatal slices, leading to the enhanced phosphorylation of Thr-75 and Ser-137 of DARPP-32, respectively. The effect of DHPG on Thr-75, but not on Ser-137, was blocked by a Cdk5-specific inhibitor, butyrolactone. In contrast, the effects of DHPG on both Thr-75 and Ser-137 were blocked by CK1-7 and IC261, specific inhibitors of CK1, suggesting that activation of Cdk5 by mGluRs requires CK1 activity. In support of this possibility, the DHPG-induced increase in Cdk5 activity, measured in extracts of neostriatal slices, was abolished by CK1-7 and IC261. Treatment of acutely dissociated neurons with DHPG enhanced voltage-dependent Ca 2؉ currents. This enhancement was eliminated by either butyrolactone or CK1-7 and was absent in DARPP-32 knockout mice. Together these results indicate that a CK1-Cdk5-DARPP-32 cascade may be involved in the regulation by mGluR agonists of Ca 2؉ channels. C yclin-dependent kinase 5 (Cdk5)͞p35 plays a variety of roles in the developing and adult nervous system (1-3). Studies of mice in which the gene encoding either Cdk5 or p35 has been disrupted have indicated that both mutants exhibit abnormalities in the laminar structure of the cerebral cortex (4, 5). Conversion of p35 to p25 by the action of calpain causes altered localization and prolonged activation of Cdk5 (6-9). Cdk5 phosphorylates DARPP-32 (dopamine-and cAMP-regulated-phosphoprotein, molecular mass 32 kDa) on Thr-75 (10, 11). Phosphorylation at Thr-34 by cAMP-dependent protein kinase (PKA) converts DARPP-32 into an inhibitor of protein phosphatase-1 (PP-1), a process that is critical for the actions of dopamine in the neostriatum (12). Phosphorylation of Thr-75 by Cdk5 converts DARPP-32 into an inhibitor of PKA, reducing phosphorylation of DARPP-32 at Thr-34, and modulating the DARPP-32͞PP-1 cascade (10).Casein kinase 1 (CK1) was one of the first serine͞threonine protein kinases to be isolated and characterized (13). It is a ubiquitous enzyme that can be found in the nucleus and the cytosol and bound to the cytoskeleton and membranes. The CK1 family consists of multiple isoforms encoded by seven distinct genes (CK1␣, ␤, ␥1, ␥2, ␥3, ␦, and ). These isoforms exhibit more than ...

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Chapter 11 Mechanisms of neuronal plasticity as analyzed at the single cell level

Cited by 11 publications

References 17 publications

Molecular and functional characteristics of MAP-2a: Ability of MAP-2a versus MAP-2b to induce stable microtubules in COS cells

Molecular and functional characteristics of MAP-2a: Ability of MAP-2a versus MAP-2b to induce stable microtubules in COS cells

Rab3A Is Required for Brain-Derived Neurotrophic Factor-Induced Synaptic Plasticity: Transcriptional Analysis at the Population and Single-Cell Levels

Regulation of cyclin-dependent kinase 5 and casein kinase 1 by metabotropic glutamate receptors

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