Isoform Specificity in the Relationship of Actin to Dendritic Spines

Kaech, Stefanie; Fischer, Maria; Doll, Thierry; Matus, Andrew

doi:10.1523/jneurosci.17-24-09565.1997

Cited by 141 publications

(99 citation statements)

References 41 publications

(49 reference statements)

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“…For all experiments in neurons, a vector using the chicken ␤-actin promoter (pBetaActin) was used to drive expression (Kaech et al, 1997). Plasmids encoding WAVE-1 were generated by PCR amplification of DNA and ligation of the restriction digested insert at sites downstream of enhanced green fluorescent protein (GFP) (Clontech).…”

Section: Plasmidsmentioning

confidence: 99%

A WAVE-1 and WRP Signaling Complex Regulates Spine Density, Synaptic Plasticity, and Memory

Soderling¹,

Guire²,

Kaech³

et al. 2007

J. Neurosci.

199

187

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The scaffolding protein WAVE-1 (Wiskott-Aldrich syndrome protein family member 1) directs signals from the GTPase Rac through the Arp2/3 complex to facilitate neuronal actin remodeling. The WAVE-associated GTPase activating protein called WRP is implicated in human mental retardation, and WAVE-1 knock-out mice have altered behavior. Neuronal time-lapse imaging, behavioral analyses, and electrophysiological recordings from genetically modified mice were used to show that WAVE-1 signaling complexes control aspects of neuronal morphogenesis and synaptic plasticity. Gene targeting experiments in mice demonstrate that WRP anchoring to WAVE-1 is a homeostatic mechanism that contributes to neuronal development and the fidelity of synaptic connectivity. This implies that signaling through WAVE-1 complexes is essential for neural plasticity and cognitive behavior.

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Section: Plasmidsmentioning

confidence: 99%

A WAVE-1 and WRP Signaling Complex Regulates Spine Density, Synaptic Plasticity, and Memory

Soderling¹,

Guire²,

Kaech³

et al. 2007

J. Neurosci.

199

187

View full text Add to dashboard Cite

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“…Actin remodeling underlies the establishment and maturation of dendritic spines, and ␤-actinimmunoreactive levels are a measure of dendritic spine density (Matus et al, 1982;Kaech et al, 1997). To begin analyzing the mechanism of estradiol action, phosphorylated cofilin was measured (Schubert et al, 2006;Carlisle et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Membrane-Initiated Estradiol Signaling Induces Spinogenesis Required for Female Sexual Receptivity

Christensen¹,

Dewing²,

Micevych³

2011

J. Neurosci.

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Estrogens have profound actions on the structure of the nervous system during development and in adulthood. One of the signature actions of estradiol is to alter the morphology of neural processes. In the hippocampus, estradiol modulates spines and cellular excitability that affect cognitive behaviors. In the hypothalamus, estradiol increases spine density in mediobasal hypothalamic nuclei that regulate reproduction. The hypothalamic arcuate nucleus (ARH), an important site for modulation of female sexual receptivity, has a sexual dimorphism in dendritic spine density that favors females. In the present study, we used both ␤-actin immunostaining and Golgi staining to visualize estradiol-induced changes in spine density in Long-Evans rats. Golgi impregnation was used to visualize spine shape, and then ␤-actin immunoreactivity was used as a semiquantitative measure of spine plasticity since actin forms the core of dendritic spines. At 4 h after estradiol treatment, both ␤-actin immunofluorescence and filopodial spines were increased (from 70.57 Ϯ 1.09% to 78.01 Ϯ 1.05%, p Ͻ 0.05). Disruption of estradiol-induced ␤-actin polymerization with cytochalasin D attenuated lordosis behavior, indicating the importance of estradiol-mediated spinogenesis for female sexual receptivity (81.43 Ϯ 7.05 to 35.00 Ϯ 11.76, p Ͻ 0.05). Deactivation of cofilin, an actin depolymerizing factor is required for spinogenesis. Membrane-initiated estradiol signaling involving the metabotropic glutamate receptor 1a was responsible for the phosphorylation and thereby deactivation of cofilin. These data demonstrate that estradiolinduced spinogenesis in the ARH is an important cellular mechanism for the regulation of female sexual behavior.

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“…A viral epitope such as the 11-amino acid peptide derived from vesicular stomatitis virus G protein (VSV-G) (Soldati and Perriard, 1991) decreases the risk that the antibody recognizing the tag crossreacts with cellular components. Insertion of this particular tag at the C terminus of different actin isoforms has been used to study their distribution relative to the endogenous actins in fibroblasts and cardiomyocytes (von Arx et al, 1995), smooth muscle, endothelial and epithelial cells (Mounier et al, 1997), and hippocampal neurons (Kaech et al, 1997). For the interpretation of these experiments it has been assumed that the tag does not interfere with the correct folding and function of the protein.…”

mentioning

confidence: 99%

Differential Epitope Tagging of Actin in TransformedDrosophilaProduces Distinct Effects on Myofibril Assembly and Function of the Indirect Flight Muscle

Brault¹,

Sauder

Reedy

et al. 1999

MBoC

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We have tested the impact of tags on the structure and function of indirect flight muscle (IFM)-specific Act88F actin by transforming mutant Drosophila melanogaster, which do not express endogenous actin in their IFMs, with tagged Act88F constructs. Epitope tagging is often the method of choice to monitor the fate of a protein when a specific antibody is not available. Studies addressing the functional significance of the closely related actin isoforms rely almost exclusively on tagged exogenous actin, because only few antibodies exist that can discriminate between isoforms. Thereby it is widely presumed that the tag does not significantly interfere with protein function. However, in most studies the tagged actin is expressed in a background of endogenous actin and, as a rule, represents only a minor fraction of the total actin. The Act88F gene encodes the only Drosophila actin isoform exclusively expressed in the highly ordered IFM. Null mutations in this gene do not affect viability, but phenotypic effects in transformants can be directly attributed to the transgene. Transgenic flies that express Act88F with either a 6x histidine tag or an 11-residue peptide derived from vesicular stomatitis virus G protein at the C terminus were flightless. Overall, the ultrastructure of the IFM resembled that of the Act88F null mutant, and only low amounts of C-terminally tagged actins were found. In contrast, expression of N-terminally tagged Act88F at amounts comparable with that of wild-type flies yielded fairly normal-looking myofibrils and partially reconstituted flight ability in the transformants. Our findings suggest that the N terminus of actin is less sensitive to modifications than the C terminus, because it can be tagged and still polymerize into functional thin filaments. INTRODUCTIONActins, a highly conserved family of cytoplasmic proteins, are among the most abundant proteins in eukaryotic cells. As a major component of the cytoskeleton, they control shape and motility in nonmuscle cells. In muscle, actin assembles into thin filaments, which together with interdigitating myosin thick filaments provide the framework for muscle contraction.Many organisms synthesize multiple isoforms of actin that are very similar in amino acid sequence even within the same cell. The differential expression of distinct actins as well as the high conservation of specific isoforms across species emphasize the functional importance of isoforms. In the case of actin, the question of how structure determines function appears to be particularly challenging. Considerable efforts have been made to understand how the different isoforms fulfill their various functions despite their extremely high sequence identity, and yet the basis of their functional diversity remains elusive. § Corresponding author. E-mail address: Schoenenberg@ ubaclu.unibas.ch. © 1999 by The American Society for Cell Biology 135Studying the specific role of a particular actin isoform has always been hampered by the difficulty of discriminating between the introduced a...

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Isoform Specificity in the Relationship of Actin to Dendritic Spines

Cited by 141 publications

References 41 publications

A WAVE-1 and WRP Signaling Complex Regulates Spine Density, Synaptic Plasticity, and Memory

A WAVE-1 and WRP Signaling Complex Regulates Spine Density, Synaptic Plasticity, and Memory

Membrane-Initiated Estradiol Signaling Induces Spinogenesis Required for Female Sexual Receptivity

Differential Epitope Tagging of Actin in TransformedDrosophilaProduces Distinct Effects on Myofibril Assembly and Function of the Indirect Flight Muscle

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