Subcellular Dynamics of Type II PKA in Neurons

Zhong, Haining; Sia, Gek Ming; Satō, Takashi; Gray, Noah W.; Mao, Tianyi; Khuchua, Zaza; Huganir, Richard L.; Svoboda, Karel

doi:10.1016/j.neuron.2009.03.013

Cited by 111 publications

(123 citation statements)

References 87 publications

Supporting

Mentioning

115

Contrasting

Order By: Relevance

“…It is tempting to speculate that recruitment of EB3 to MAP2-decorated mi- Illenberger et al, 1996;Lim and Halpain, 2000;Zhong et al, 2009). Our data also indicate that EB3 has a higher affinity for dephosphorylated MAP2.…”

Section: Stable Map2-decorated Microtubules Trap Eb3mentioning

confidence: 58%

NMDA Receptor Activation Suppresses Microtubule Growth and Spine Entry

Kapitein

Yau²,

Gouveia³

et al. 2011

J. Neurosci.

106

105

View full text Add to dashboard Cite

Dynamic microtubules are important to maintain neuronal morphology and function, but whether neuronal activity affects the organization of dynamic microtubules is unknown. Here, we show that a protocol to induce NMDA-dependent long-term depression (LTD) rapidly attenuates microtubule dynamics in primary rat hippocampal neurons, removing the microtubule-binding protein EB3 from the growing microtubule plus-ends in dendrites. This effect requires the entry of calcium and is mediated by activation of NR2B-containing NMDA-type glutamate receptor. The rapid NMDA effect is followed by a second, more prolonged response, during which EB3 accumulates along MAP2-positive microtubule bundles in the dendritic shaft. MAP2 is both required and sufficient for this activity-dependent redistribution of EB3. Importantly, NMDA receptor activation suppresses microtubule entry in dendritic spines, whereas overexpression of EB3-GFP prevents NMDA-induced spine shrinkage. These results suggest that short-lasting and long-lasting changes in dendritic microtubule dynamics are important determinants for NMDA-induced LTD.

show abstract

Section: Stable Map2-decorated Microtubules Trap Eb3mentioning

confidence: 58%

NMDA Receptor Activation Suppresses Microtubule Growth and Spine Entry

Kapitein

Yau²,

Gouveia³

et al. 2011

J. Neurosci.

106

105

View full text Add to dashboard Cite

show abstract

“…It is interesting to note that apart from AKAP79/150 there are numerous other AKAPs that are expressed in neurons (68 -70) that may take on distinct or partially overlapping roles in compartmentalizing PKA activity. Recently, the non-AKAP microtubule-binding protein MAP2 was identified as a major anchoring protein for PKA (type II) in neurons (71). It was found to be crucial for regulating trafficking of the catalytic subunit between neuronal dendrites and spines.…”

Section: Discussionmentioning

confidence: 99%

Imaging CREB Activation in Living Cells

Friedrich¹,

Aramuni²,

Mank

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

The Ca 2؉ -and cAMP-responsive element-binding protein (CREB) and the related ATF-1 and CREM are stimulus-inducible transcription factors that link certain forms of cellular activity to changes in gene expression. They are attributed to complex integrative activation characteristics, but current biochemical technology does not allow dynamic imaging of CREB activation in single cells. Using fluorescence resonance energy transfer between mutants of green fluorescent protein we here develop a signal-optimized genetically encoded indicator that enables imaging activation of CREB due to phosphorylation of the critical serine 133. The indicator of CREB activation due to phosphorylation (ICAP) was used to investigate the role of the scaffold and anchoring protein AKAP79/ 150 in regulating signal pathways converging on CREB. We show that disruption of AKAP79/150-mediated protein kinase A anchoring or knock-down of AKAP150 dramatically reduces the ability of protein kinase A to activate CREB. In contrast, AKAP79/150 regulation of CREB via L-type channels may only have minor importance. ICAP allows dynamic and reversible imaging in living cells and may become useful in studying molecular components and cell-type specificity of activity-dependent gene expression.Changes in gene expression are a hallmark of long term adaptive processes in many cell types and are crucially involved in the conversion from short term to long term plasticity of neuronal circuits. Transcription factors of the cAMP-responsive element-binding protein (CREB) 3 family have a prominent role in affecting such processes in various tissues and cell types (1-5). The family includes the homologous proteins CREB (6, 7), ATF-1 (8), and CREM (9). Upon stimulation a number of kinases phosphorylate the critical serine 133 residue within the kinase-inducible domain (KID) of CREB and the related transcription factors ATF-1 and CREM. Serine 133 phosphorylation leads to the recruitment of transcriptional coactivators, like CBP or its paralogue P300 that bind to the KID via the KID interaction domain (KIX) (10, 11). Interaction of CREB with CBP/P300 results in the assembly of the RNA-polymerase complex and in the expression of a large number of genes. Serine 133 phosphorylation is generally accepted as a key event in transcriptional regulation necessary although not in all cases sufficient, for CREB-mediated gene expression. Many studies focused on examining spatiotemporal profiles of CREB activation in cells and tissues by immunostainings using antibodies specific for serine 133 phosphorylation (for a selection, see Refs. 12-17). Few attempts were made to develop assays for real time imaging of CREB, a prerequisite for thorough analysis of the physiological events and the molecular dissection of signaling cascades converging on CREB (18,19). Genetically encoded indicators based on mutants of the green fluorescent protein (GFP) have become valuable tools to monitor spatiotemporal patterns of biochemical signals in live cells (20 -25). A number of indicators have ...

show abstract

“…In utero electroporation was performed as described previously (Gray et al, 2006;Zhong et al, 2009) at E15.5 using an electroporator (CUY21, BEX) with five 100 ms pulses of 38 V. cDNA encoding the Cre recombinase were put into a vector with a CAG promoter, an intron, and a WHP post-transcriptional response element. For in utero viral infection, 0.5-1 l of diluted AAV9 or AAV 2/1 virus that directs the expression of Cre under a CAG promoter was injected to the lateral ventricles of E15.5 mouse embryos using a surgical procedure essentially identical to that for in utero electroporation, with the omission of electroporation.…”

Section: Generation Of Psd-95-enabled and Psd-95-crenabled Micementioning

confidence: 99%

Live Imaging of Endogenous PSD-95 Using ENABLED: A Conditional Strategy to Fluorescently Label Endogenous Proteins

et al. 2014

View full text Add to dashboard Cite

Stoichiometric labeling of endogenous synaptic proteins for high-contrast live-cell imaging in brain tissue remains challenging. Here, we describe a conditional mouse genetic strategy termed endogenous labeling via exon duplication (ENABLED), which can be used to fluorescently label endogenous proteins with near ideal properties in all neurons, a sparse subset of neurons, or specific neuronal subtypes. We used this method to label the postsynaptic density protein PSD-95 with mVenus without overexpression side effects. We demonstrated that mVenus-tagged PSD-95 is functionally equivalent to wild-type PSD-95 and that PSD-95 is present in nearly all dendritic spines in CA1 neurons. Within spines, while PSD-95 exhibited low mobility under basal conditions, its levels could be regulated by chronic changes in neuronal activity. Notably, labeled PSD-95 also allowed us to visualize and unambiguously examine otherwiseunidentifiable excitatory shaft synapses in aspiny neurons, such as parvalbumin-positive interneurons and dopaminergic neurons. Our results demonstrate that the ENABLED strategy provides a valuable new approach to study the dynamics of endogenous synaptic proteins in vivo.

show abstract

Subcellular Dynamics of Type II PKA in Neurons

Cited by 111 publications

References 87 publications

NMDA Receptor Activation Suppresses Microtubule Growth and Spine Entry

NMDA Receptor Activation Suppresses Microtubule Growth and Spine Entry

Imaging CREB Activation in Living Cells

Live Imaging of Endogenous PSD-95 Using ENABLED: A Conditional Strategy to Fluorescently Label Endogenous Proteins

Contact Info

Product

Resources

About