Synapses: Changing the size of dendritic spines

Interactions between an enzyme kinase, an ion channel and cytoskeletal proteins maintain the structure of synapses involved in memory formation.

Sep 7, 2023

https://doi.org/10.7554/eLife.91566

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Takeo Saneyoshi

Department of Pharmacology, Kyoto University Graduate School of Medicine, Japan

Neurons connect to one another at sites called synapses, which are essential for creating and storing memories. Synapses are also dynamic and can undergo structural changes, such as increasing in size, that modify their strength. Structures called dendritic spines – which protrude from the dendrites that extend from the cell body of a neuron – have an important role in synapses as they contain an actin cytoskeleton that allows them to change shape and size.

An enzyme called CaMKII (which is short for Ca²⁺/calmodulin-dependent protein kinase II) also has a central role in the functional and structural changes that take place in dendritic spines during memory formation. During synapse strengthening, an influx of calcium ions through a receptor known as NMDAR, triggers the activation of CaMKII: the calcium ions form a complex with a messenger protein called calmodulin, and this complex binds to the regulatory segment of CaMKII. This influences the dynamics of the actin cytoskeleton and hence determines the size of the synapses. However, the details of this process – especially the link between CaMKII and the actin cytoskeleton at the molecular level – are not fully understood.

One candidate for linking CaMKII to the actin cytoskeleton is alpha-actinin-2, a structural protein that crosslinks actin filaments (Walikonis et al., 2001). This protein can anchor and organize other proteins in the dendritic spine, such as CaMKII (Robison et al., 2005), the receptor NMDAR (Wyszynski et al., 1997), and certain channel proteins that allow calcium ions to enter cells (Hall et al., 2013). Now, in eLife, Matthew Gold from University College London and colleagues – including Ashton Curtis and Jian Zhu as joint first authors – report new insights into the molecular interactions between CaMKII, alpha-actinin-2, and a subunit of NMDAR called GluN2B (Curtis et al., 2023; Figure 1).

Figure 1

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Molecular interactions and synapses.

(A) In the naïve synapse, the only protein interactions are between actin filaments (blue) and alpha-actinin-2, a structural protein that crosslinks actin filaments. In particular, the enzyme CaMKII (hexagon shape) is inactive, and a messenger protein called calmodulin (CaM) does not interact with anything else. (B) When the NMDAR receptor in the cell membrane is activated by the neurotransmitter glutamate, calcium ions (Ca²⁺; solid pink circles) enter the cell. The ions form a complex with CaM, and this complex activates the enzyme CaMKII, which subsequently interacts with the GluN2B subunits in the intracellular region of the NMDAR receptor. Crosslinking of the actin filaments also causes the dendritic spine to increase in size. (C) In the enlarged synapse, the interaction between GluN2B, CaMKII and alpha-actinin-2 continues, even in the absence of Ca²⁺/CaM, and the enzymatic activity of CaMKII is maintained by binding to GluN2B. The complex formed by GluN2B, CaMKII and alpha-actinin-2 also associates with actin filaments to maintain the synaptic structure.

Using a proximity labeling assay method – which enables molecular interactions to be detected in situ – the researchers showed that CaMKII interacts with alpha-actinin-2 when the NMDAR receptor is stimulated, leading to an enlargement of the dendritic spine. The interaction takes place at a domain in alpha-actinin-2 called EF hands 1–4, which is known to bind calcium ions. Furthermore, when this domain was overexpressed, stimulation of the NMDAR receptor did not result in dendritic spine enlargement, suggesting that interaction via the EF1-4 domain has a critical role in structural changes to the dendritic spine.

Affinity and structural experiments showed that alpha-actinin-2 binds to the CaMKII enzyme at the same regulatory domain segment as calmodulin (which also contains EF hands) does. However, Curtis et al. found that the EF hands of alpha-actinin-2 clashed with one of the domains in the enzyme. Consistent with this, the inactive form of the enzyme cannot bind to alpha-actinin-2 via its regulatory domain segment. However, the interaction between the CaMKII enzyme and alpha-actinin-2 in the dendritic spine increased four hours after stimulation of the NMDAR receptor: this led Curtis et al. to hypothesize that, following stimulation, the configuration of the enzyme changes when it interacts with a subunit of the NMDAR receptor called GluN2B, and these changes enable alpha-actinin-2 to fully access the enzyme.

A pull-down assay – which detects interactions between proteins – revealed that GluN2B enhances the interaction between the CaMKII enzyme and alpha-actinin-2, independently of the concentration of calcium ions (Ca²⁺). This indicates that GluN2B enhances the binding of CaMKII with alpha-actinin-2 even in the presence of Ca²⁺/calmodulin, although the EF hands of alpha-actinin-2 have a lower affinity for CaMKII than the EF hands of Ca²⁺/calmodulin. This provides important insights into a reorganization process for the molecular architecture of synapses.

Taken together, the findings suggest that the tripartite interactions of CaMKII, GluN2B, and alpha-actinin-2 crosslink a membrane channel to the cytoskeleton beneath the synapse, and there are plenty of empty binding sites that other proteins can bind to. Moreover, the fact that certain interactions would generate autonomous enzymatic activity in CaMKII (Bayer et al., 2001; Saneyoshi et al., 2019) suggest a possible role in biochemical signalling for synaptic memory.

Recent evidence indicates that the CaMKII and GluN2B in synapses can undergo liquid–liquid phase separation, a process that involves biomolecules separating into distinct components within a cell (Hosokawa et al., 2021; Yasuda et al., 2022). Actin bundling can also be modulated by liquid–liquid phase separation in synapses (Chen et al., 2023), so it would be interesting to explore if this process has a role in changing and/or maintaining the structure of dendritic spines.

References

(2001) Interaction with the NMDA receptor locks CaMKII in an active conformation
Nature 411:801–805.
https://doi.org/10.1038/35081080
- PubMed
- Google Scholar
1. Chen X
2. Jia B
3. Zhu S
4. Zhang M
(2023) Phase separation-mediated actin bundling by the postsynaptic density condensates
eLife 12:e84446.
https://doi.org/10.7554/eLife.84446
- PubMed
- Google Scholar
1. Curtis AJ
2. Zhu J
3. Penny CJ
4. Gold MG
(2023) Molecular basis of interactions between CaMKII and α-actinin-2 that underlie dendritic spine enlargement
eLife 12:e85008.
https://doi.org/10.7554/eLife.85008
- PubMed
- Google Scholar
1. Hall DD
2. Dai S
3. Tseng PY
4. Malik Z
5. Nguyen M
6. Matt L
7. Schnizler K
8. Shephard A
9. Mohapatra DP
10. Tsuruta F
11. Dolmetsch RE
12. Christel CJ
13. Lee A
14. Burette A
15. Weinberg RJ
16. Hell JW
(2013) Competition between α-actinin and Ca²⁺-calmodulin controls surface retention of the L-type Ca²⁺ channel Ca(V)1.2
Neuron 78:483–497.
https://doi.org/10.1016/j.neuron.2013.02.032
- PubMed
- Google Scholar
1. Hosokawa T
2. Liu PW
3. Cai Q
4. Ferreira JS
5. Levet F
6. Butler C
7. Sibarita JB
8. Choquet D
9. Groc L
10. Hosy E
11. Zhang M
12. Hayashi Y
(2021) CaMKII activation persistently segregates postsynaptic proteins via liquid phase separation
Nature Neuroscience 24:777–785.
https://doi.org/10.1038/s41593-021-00843-3
- PubMed
- Google Scholar
1. Robison AJ
2. Bass MA
3. Jiao Y
4. MacMillan LB
5. Carmody LC
6. Bartlett RK
7. Colbran RJ
(2005) Multivalent interactions of calcium/calmodulin-dependent protein kinase II with the postsynaptic density proteins NR2B, densin-180, and alpha-actinin-2
The Journal of Biological Chemistry 280:35329–35336.
https://doi.org/10.1074/jbc.M502191200
- PubMed
- Google Scholar
1. Saneyoshi T
2. Matsuno H
3. Suzuki A
4. Murakoshi H
5. Hedrick NG
6. Agnello E
7. O’Connell R
8. Stratton MM
9. Yasuda R
10. Hayashi Y
(2019) Reciprocal Activation within a Kinase-Effector Complex Underlying Persistence of Structural LTP
Neuron 102:1199–1210.
https://doi.org/10.1016/j.neuron.2019.04.012
- PubMed
- Google Scholar
(2001) Densin-180 forms a ternary complex with the α-subunit of Ca2+/calmodulin-dependent protein kinase II and α-actinin
The Journal of Neuroscience 21:423–433.
https://doi.org/10.1523/JNEUROSCI.21-02-00423.2001
- PubMed
- Google Scholar
1. Wyszynski M
2. Lin J
3. Rao A
4. Nigh E
5. Beggs AH
6. Craig AM
7. Sheng M
(1997) Competitive binding of α-actinin and calmodulin to the NMDA receptor
Nature 385:439–442.
https://doi.org/10.1038/385439a0
- PubMed
- Google Scholar
(2022) CaMKII: a central molecular organizer of synaptic plasticity, learning and memory
Nature Reviews Neuroscience 23:666–682.
https://doi.org/10.1038/s41583-022-00624-2
- PubMed
- Google Scholar

Article and author information

Author details

Takeo Saneyoshi

Takeo Saneyoshi is in the Department of Pharmacology, Kyoto University Graduate School of Medicine, Kyoto, Japan

For correspondence
saneyoshi.takeo.3v@kyoto-u.ac.jp

Competing interests
No competing interests declared

"This ORCID iD identifies the author of this article:" 0000-0003-2361-3339

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Version of Record published: September 7, 2023 (version 1)

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