**4. Spine plasticity**

In young adolescent mice (1-month old), within a 2 week period of time, 13–20% of total spines were eliminated with 5–8% formed in the barrel, motor and frontal cortices, and this imbalance led to a significant spine loss in many brain regions. However, in the adult mice (4–6 months old), 3–5% of spines were eliminated and formed in 2 weeks [18]. Most evidences support that dendritic spines are stable in the adulthood [37, 38]. Grutzendler et al. reported that spines in the primary visual cortex of young adolescence have a turnover rate of 27% per month but this rate dropped to only 4% in the adult [15]. In contrast, Trachtenberg et al. found that adult spines are highly dynamic with about 20% turnover per day in the mouse barrel cortex [16]. This discrepancy is likely due to differences in the methodology in that the use of cranial window in the latter study triggered inflammatory responses in the brain which resulted in elevated turnover rates. It should be pointed out that spine turnover rate differs in various brain regions. For example, Holtmaat et al. reported that spines turn over more slowly (both generation and elimination) in the visual cortex than in the somatosensory cortex, with the fraction of transient spines (lifetime ≤ 4 days) also lower in visual

**3. Signaling events during spine formation and maintenance**

Motility of dendritic spines is regulated by the dynamic balance between G-actin and F-actin [39]. F-actin consists of two pools, a large dynamic pool in the tip of the spine head and a small stable pool in the base of spine [6] [40]. With LTP induction, the stable F-actin is severed into short segments and reorganized to expand the spine [41]. Thus, the dynamics of actin cytoskeleton controls dendritic spine morphological remodeling and plenty of signaling mol-

Spine morphology is regulated by actin binding and cytoskeleton proteins. Drebrin was the first identified to modify dendritic spines since overexpression of drebrin in cultured neurons increased the length of spines [45]. Spines in the drebrin knockout mice exhibited normal morphology but altered plasticity [46]. Takahashi et al. reported that drebrin entered filopodia and formed an actin filament cluster to recruit postsynaptic components (including scaffolding protein PSD95), and this process enables the transition from filopodia to mature spines. Based on this observation, filopodia are classified into two types, an immature diffuse-type and a mature cluster-type. A filopodium with a drebrin cluster, whose maximum intensity was higher than twice the average intensity of the filopodium, was classified as a cluster-type filopodium. Otherwise, it was classified as a diffuse-type filopodium. The clustertype filopodia were likely to be converted to mature spines [47]. In addition, overexpression of drebrin in neurons caused F-actin to accumulate in the growth cone, whereas knockdown of drebrin reduced F-actin level [48]. Drebrin binds to F-actin to generate thick bundles of F-actin [49], and drebrin also competes with other actin binding proteins such as ADF/cofilin

Besides drebrin, other actin-binding proteins including myosin II, Abi-1 and spinophilin regulate actin polymerization in the dendritic spines. Myosin II belongs to the family of molecular motors which is highly expressed in dendritic spines, and regulates dendritic spine morphology and

cortex [17].

ecules participate in this process [42–44].

150 Neuroplasticity - Insights of Neural Reorganization

which depolymerizes F-actin [50, 51].

Spine plasticity may be exhibited in two forms—changes in spine density and spine dimension. Change in spine density reflects modification of connection density between the presynaptic and postsynaptic neurons, which happens most commonly during brain development (increase, decrease/pruning) and aging/degeneration (decrease). Changes in spine morphology/dimension, especially the size of spine head, have been widely reported, and are believed to be associated with changes in the strength of synapses that reside on these altered spines. In this chapter, we will focus on alterations of spine dimension.

#### **4.1. Changes in spine morphology**

Due to the heterogeneity of spine size/morphology, the most convincing way to demonstrate that spine morphology is altered is to compare the same set of spines before and after a manipulation, such as synaptic plasticity-inducing stimuli in brain slices or learning *in vivo*. By using time-lapse two-photon imaging on the same set of spines, Yang et al. found new spines were formed after *in vivo* experience in the form of sensory or motor, and a fraction of these newly appeared spines persisted for months after the experience. More importantly, of the appearance of these new spines is specifically related to the *in vivo* experience or training [66]. In addition, Hayashi-Takagi et al. showed that motor learning on rotarod led to an enhanced *Arc* signaling, together with an expansion of a subset of spines in the motor cortex. By expressing a photoactivatable GTPase Rac1 in spines, they further showed that prolonged photo-activation of Rac1 resulted in reversal of spine expansion and loss of motor memory. This is a striking demonstration that potentiated synapses and enlarged spines are likely the underlying biological substrates of stored memory and formed memory can be erased by reversing these changes [67].

Many studies have examined spine modifications with the induction of long-term potentiation (LTP) and long-term depression (LTD), since this procedure allows the examination of the same set spines associated with fast, large and long-lasting changes in synaptic strength. These two forms of synaptic plasticity refer to the increase and decrease in synaptic strength respectively, and are generally regarded as the cellular basis of synaptic modifications underlying developmental remodeling of neuronal connections, and learning and memory function in the adult brain [68]. When studied in brain slices (acute or organotypic culture), LTP or LTD is induced by stimulation of the presynaptic inputs with distinct patterns. In some studies, changes in the synaptic strength were also monitored, and thus changes in synaptic function and spine morphology can be related to each other in the same set of spines/synapse, or even a single synapse/spine [34, 69–71]. In general, spines exhibit the capacity of bi-directional changes in that spine enlargement is observed with LTP while spine shrinkage with LTD [72–74] (**Figure 2**). Either uncaging of caged glutamate onto a single spine [75] or electrical stimulation of a population of synapses [70, 71] had confirmed the above observations. Uncaging of glutamate directly enhances postsynaptic AMPAR function, and since it bypasses presynaptic release, and thus has provided the unambiguous evidence that postsynaptic changes can underlie the expression of LTP [69, 75]. These observations further indicate that morphological and functional changes are likely driven by the same stimuli or process (see below).

reported that the number of AMPAR in spines is of a large range. In addition, mushroom spines are enriched with AMPARs, compared to the low distribution of them in the thin spines and filopodia. These observations support a strong relationship between number of AMPAR and volume of spines [77]. In addition, it provides direct evidence that mushroom spines are func-

**Figure 2.** Bi-directional modification of spine size by synaptic plasticity. (A) Recording, synaptic stimulation and fluorescence imaging of the same set of synapses and spines in acute hippocampal slices. (B) Spine enlargement associated with LTP induction by theta burst stimulation, this enlargement is persistent and long-lasting. In addition, enlargement was restricted to spines close (upper) but not far away from the stimulation site (lower). (C) Change in spine volume before and after LTP induction and its requirement of activation of NMDARs since it was prevented by bath application of NMDAR antagonists APV and MK-801. (D) Increase in synaptic strength (EPSPs) as a result of LTP induction. LTP occurred in two phases, an initial rapid increase (indicated by 2 on the plot) and a slower gradual increase (indicated by 3). LTP was also sensitive to NMDAR blockade. (E) Spine shrinkage associated with LTD, it is persistent and long-lasting. (F) Shrinkage of spines occurred to those spines close to, but not to those far away from the stimulating electrode, or those had not received any stimulation. Compared to the almost instantaneous enlargement of spine heads after LTP, spine shrinkage develops slowly and takes much longer to reach a plateau. (G) Low frequency stimulation led

Dendritic Spine Modifications in Brain Physiology http://dx.doi.org/10.5772/intechopen.73326 153

Synapses can change their strength by the activity patterns that they receive, and this modification allows synaptic strength to be adjusted to better suit the need for adaptation. Originally put forwarded by Donald Hebb and later adopted as "fire together, wire together" model, the current model of synaptic modification states that neurons sharing spike activity have increased connections between these two partners [78]. After the discovery of LTP in 1973, this activity-driven increase in synaptic strength has been extensively studied, both in acute brain slices and *in vivo* [68]; and has been used widely as a tool to induce synaptic modification in order to study the associated processes. Various molecules have been identified to be required for the induction and expression of LTP [68]. In general, Ca2+ entry or elevated intracellular Ca2+ concentration is required to convert electrical activity into intracellular signaling that determines the direction of synaptic changes. Usually a large but transient elevation in

tionally mature while thin spines and filopodia are not.

**4.2. Synaptic plasticity**

to depression of EPSPs. Taken from [34, 71].

In general, there is a good correlation between the strength of a given synapse (measured by electrophysiological responses) and the size of spine. Electrophysiological responses are further determined by the number/density of AMPA receptors at a given synapse. Takumi et al. found a linear relationship between AMPAR density and the diameter of PSD [76]. Matsuzaki et al.

**Figure 2.** Bi-directional modification of spine size by synaptic plasticity. (A) Recording, synaptic stimulation and fluorescence imaging of the same set of synapses and spines in acute hippocampal slices. (B) Spine enlargement associated with LTP induction by theta burst stimulation, this enlargement is persistent and long-lasting. In addition, enlargement was restricted to spines close (upper) but not far away from the stimulation site (lower). (C) Change in spine volume before and after LTP induction and its requirement of activation of NMDARs since it was prevented by bath application of NMDAR antagonists APV and MK-801. (D) Increase in synaptic strength (EPSPs) as a result of LTP induction. LTP occurred in two phases, an initial rapid increase (indicated by 2 on the plot) and a slower gradual increase (indicated by 3). LTP was also sensitive to NMDAR blockade. (E) Spine shrinkage associated with LTD, it is persistent and long-lasting. (F) Shrinkage of spines occurred to those spines close to, but not to those far away from the stimulating electrode, or those had not received any stimulation. Compared to the almost instantaneous enlargement of spine heads after LTP, spine shrinkage develops slowly and takes much longer to reach a plateau. (G) Low frequency stimulation led to depression of EPSPs. Taken from [34, 71].

reported that the number of AMPAR in spines is of a large range. In addition, mushroom spines are enriched with AMPARs, compared to the low distribution of them in the thin spines and filopodia. These observations support a strong relationship between number of AMPAR and volume of spines [77]. In addition, it provides direct evidence that mushroom spines are functionally mature while thin spines and filopodia are not.

#### **4.2. Synaptic plasticity**

morphology/dimension, especially the size of spine head, have been widely reported, and are believed to be associated with changes in the strength of synapses that reside on these altered

Due to the heterogeneity of spine size/morphology, the most convincing way to demonstrate that spine morphology is altered is to compare the same set of spines before and after a manipulation, such as synaptic plasticity-inducing stimuli in brain slices or learning *in vivo*. By using time-lapse two-photon imaging on the same set of spines, Yang et al. found new spines were formed after *in vivo* experience in the form of sensory or motor, and a fraction of these newly appeared spines persisted for months after the experience. More importantly, of the appearance of these new spines is specifically related to the *in vivo* experience or training [66]. In addition, Hayashi-Takagi et al. showed that motor learning on rotarod led to an enhanced *Arc* signaling, together with an expansion of a subset of spines in the motor cortex. By expressing a photoactivatable GTPase Rac1 in spines, they further showed that prolonged photo-activation of Rac1 resulted in reversal of spine expansion and loss of motor memory. This is a striking demonstration that potentiated synapses and enlarged spines are likely the underlying biological substrates of stored memory and formed memory can be erased by

Many studies have examined spine modifications with the induction of long-term potentiation (LTP) and long-term depression (LTD), since this procedure allows the examination of the same set spines associated with fast, large and long-lasting changes in synaptic strength. These two forms of synaptic plasticity refer to the increase and decrease in synaptic strength respectively, and are generally regarded as the cellular basis of synaptic modifications underlying developmental remodeling of neuronal connections, and learning and memory function in the adult brain [68]. When studied in brain slices (acute or organotypic culture), LTP or LTD is induced by stimulation of the presynaptic inputs with distinct patterns. In some studies, changes in the synaptic strength were also monitored, and thus changes in synaptic function and spine morphology can be related to each other in the same set of spines/synapse, or even a single synapse/spine [34, 69–71]. In general, spines exhibit the capacity of bi-directional changes in that spine enlargement is observed with LTP while spine shrinkage with LTD [72–74] (**Figure 2**). Either uncaging of caged glutamate onto a single spine [75] or electrical stimulation of a population of synapses [70, 71] had confirmed the above observations. Uncaging of glutamate directly enhances postsynaptic AMPAR function, and since it bypasses presynaptic release, and thus has provided the unambiguous evidence that postsynaptic changes can underlie the expression of LTP [69, 75]. These observations further indicate that morphological and functional changes are likely driven

In general, there is a good correlation between the strength of a given synapse (measured by electrophysiological responses) and the size of spine. Electrophysiological responses are further determined by the number/density of AMPA receptors at a given synapse. Takumi et al. found a linear relationship between AMPAR density and the diameter of PSD [76]. Matsuzaki et al.

spines. In this chapter, we will focus on alterations of spine dimension.

**4.1. Changes in spine morphology**

152 Neuroplasticity - Insights of Neural Reorganization

reversing these changes [67].

by the same stimuli or process (see below).

Synapses can change their strength by the activity patterns that they receive, and this modification allows synaptic strength to be adjusted to better suit the need for adaptation. Originally put forwarded by Donald Hebb and later adopted as "fire together, wire together" model, the current model of synaptic modification states that neurons sharing spike activity have increased connections between these two partners [78]. After the discovery of LTP in 1973, this activity-driven increase in synaptic strength has been extensively studied, both in acute brain slices and *in vivo* [68]; and has been used widely as a tool to induce synaptic modification in order to study the associated processes. Various molecules have been identified to be required for the induction and expression of LTP [68]. In general, Ca2+ entry or elevated intracellular Ca2+ concentration is required to convert electrical activity into intracellular signaling that determines the direction of synaptic changes. Usually a large but transient elevation in Ca2+ concentration induces LTP while a small but much longer elevation in Ca2+ concentration results in LTD [79]. This Ca2+ entry can be through opening NMDA receptors, voltagegated Ca2+ channels or metabotropic glutamate receptors [68]. Increase in postsynaptic kinase activity is usually required for LTP while phosphatase activity required for LTD. After a long debate, it is now generally agreed that synaptic modifications are expressed in the postsynaptic neurons, except in a few specific cases (such as mossy fiber LTP in the hippocampal CA3 region) [80, 81]. Postsynaptic changes involve the translocation of AMPARs and their phosphorylation state, in that LTP is associated with translocation of AMPARs to synapse and/ or increased phosphorylation of AMPARs while LTD with endocytosis of AMPARs and/or dephosphorylation of AMPARs [82–85].

#### **4.3. Relationship between spine and synapse plasticity**

Since both increase in the synaptic strength and enlargement of dendritic spine occur with LTP, an obvious question is whether changes in synaptic physiology/function are casually related to changes in spine morphology/structure. More specifically, are these two processes driven by the same initial process? Does the occurrence or persistence of one process require the occurrence/presence of the other? It is now well established that influx of Ca2+ through synaptic NMDARs during LTP induction drives AMPAR phosphorylation and/or insertion [68], and polymerization of action filaments inside spines which drives enlargement of spine heads [74] (**Figure 3**). Thus, the initial changes in function and structure are driven by the same signaling process. This initial increase in synaptic response and spine volume occurs very rapidly (less than 1 min) [71].

Dendritic spine heads accumulate F-actin during the rapid expansion phase of synaptic modification. Potentiation of single synapse/spine with uncaging of glutamate led to a significant expansion of the spine head and a shortening and widening of the spine neck [86, 87]. Spine expansion takes place rapidly after LTP, as fast as it can be measured (~ 20 sec after LTP induction) [71]. F-actin concentration inside the spine head rises, together with the entry of actinsevering, actin-depolymerizing/−polymerizing, actin-capping proteins, while actin-stabilizing proteins leave the spines [6, 39, 86, 88–90]. Actin-depolymerizing agent, cofilin, is highly elevated in spines during this initial process [89]. Interestingly, unlike the expression of LTP, this initial spine expansion did not require postsynaptic exocytosis or PKA signaling [71], suggesting the involvement of different signaling pathways in spine enlargement than that supports LTP. After this initial rapid expansion, the next phase of events lasts up to 1 h, with spine head volume decreased from the initial increase, but still larger than the pre-LTP baseline. In addition, the total actin concentration in the spine may drop to the baseline level [89].

and Linden demonstrated that LTD expression and spine changes in cerebellar Purkinje cells could be induced independently of each other, and induction of one did not affect the other [92]. Wang et al. reported that trafficking of AMPARs to and away from PSDs was activityindependent and not associated with alterations in spine size. The significance of this finding

**Figure 3.** Time-dependent reversal of synaptic and spine modification. (A) Spine shrinkage induced by low frequency stimulation (LFS) can be readily reversed by subsequent high frequency stimulation (HFS). (B) Spine enlargement induced by HFS is also readily reversed by subsequent LFS. (C) Spine enlargement induced by TBP can be reversed by LFS only if LFS is given within a time window of about 15 min after TBP. (D) this critical reversal window also holds for reversing TBP-induced LTP. (E) LFS given outside this reversal window does not affect spine size. (F) LFS given outside

Dendritic Spine Modifications in Brain Physiology http://dx.doi.org/10.5772/intechopen.73326 155

One interesting and important feature of synaptic modification is its reversibility. This reversibility is defined by reversal of synaptic modification after its induction [34, 93]. There are a few aspects to this reversal: (1) reversal applies to both LTP and LTD, and spine enlargement and spine shrinkage [34, 71] (**Figure 4**). More specifically, low frequency stimulation reverses LTP and spine enlargement, while high frequency stimulation reverses LTD

requires further investigation [70].

the reversal window does not reverse LTP either. Taken from [34, 71].

LTD is associated with the shrinkage of spines and removal of synaptic AMPARs via internalization [34, 36, 39]. During the induction of LTD with low frequency synaptic stimulation, Ca2+ influx through the activated NMDARs is required for both LTD and spine shrinkage [34, 70]. Ca2+ entry through synaptic NMDARs leads to the activation of calcineurin which is also required for both LTD and spine shrinkage, while activation of protein phosphatase 2A is required for LTD expression but not spine shrinkage, while elevated cofilin activity is required for spine shrinkage but not LTD [34, 39, 70, 91]. Consistent with the above conclusion, Sdrulla

Ca2+ concentration induces LTP while a small but much longer elevation in Ca2+ concentration results in LTD [79]. This Ca2+ entry can be through opening NMDA receptors, voltagegated Ca2+ channels or metabotropic glutamate receptors [68]. Increase in postsynaptic kinase activity is usually required for LTP while phosphatase activity required for LTD. After a long debate, it is now generally agreed that synaptic modifications are expressed in the postsynaptic neurons, except in a few specific cases (such as mossy fiber LTP in the hippocampal CA3 region) [80, 81]. Postsynaptic changes involve the translocation of AMPARs and their phosphorylation state, in that LTP is associated with translocation of AMPARs to synapse and/ or increased phosphorylation of AMPARs while LTD with endocytosis of AMPARs and/or de-

Since both increase in the synaptic strength and enlargement of dendritic spine occur with LTP, an obvious question is whether changes in synaptic physiology/function are casually related to changes in spine morphology/structure. More specifically, are these two processes driven by the same initial process? Does the occurrence or persistence of one process require the occurrence/presence of the other? It is now well established that influx of Ca2+ through synaptic NMDARs during LTP induction drives AMPAR phosphorylation and/or insertion [68], and polymerization of action filaments inside spines which drives enlargement of spine heads [74] (**Figure 3**). Thus, the initial changes in function and structure are driven by the same signaling process. This initial increase in synaptic response and spine volume occurs

Dendritic spine heads accumulate F-actin during the rapid expansion phase of synaptic modification. Potentiation of single synapse/spine with uncaging of glutamate led to a significant expansion of the spine head and a shortening and widening of the spine neck [86, 87]. Spine expansion takes place rapidly after LTP, as fast as it can be measured (~ 20 sec after LTP induction) [71]. F-actin concentration inside the spine head rises, together with the entry of actinsevering, actin-depolymerizing/−polymerizing, actin-capping proteins, while actin-stabilizing proteins leave the spines [6, 39, 86, 88–90]. Actin-depolymerizing agent, cofilin, is highly elevated in spines during this initial process [89]. Interestingly, unlike the expression of LTP, this initial spine expansion did not require postsynaptic exocytosis or PKA signaling [71], suggesting the involvement of different signaling pathways in spine enlargement than that supports LTP. After this initial rapid expansion, the next phase of events lasts up to 1 h, with spine head volume decreased from the initial increase, but still larger than the pre-LTP baseline. In addi-

tion, the total actin concentration in the spine may drop to the baseline level [89].

LTD is associated with the shrinkage of spines and removal of synaptic AMPARs via internalization [34, 36, 39]. During the induction of LTD with low frequency synaptic stimulation, Ca2+ influx through the activated NMDARs is required for both LTD and spine shrinkage [34, 70]. Ca2+ entry through synaptic NMDARs leads to the activation of calcineurin which is also required for both LTD and spine shrinkage, while activation of protein phosphatase 2A is required for LTD expression but not spine shrinkage, while elevated cofilin activity is required for spine shrinkage but not LTD [34, 39, 70, 91]. Consistent with the above conclusion, Sdrulla

phosphorylation of AMPARs [82–85].

154 Neuroplasticity - Insights of Neural Reorganization

very rapidly (less than 1 min) [71].

**4.3. Relationship between spine and synapse plasticity**

**Figure 3.** Time-dependent reversal of synaptic and spine modification. (A) Spine shrinkage induced by low frequency stimulation (LFS) can be readily reversed by subsequent high frequency stimulation (HFS). (B) Spine enlargement induced by HFS is also readily reversed by subsequent LFS. (C) Spine enlargement induced by TBP can be reversed by LFS only if LFS is given within a time window of about 15 min after TBP. (D) this critical reversal window also holds for reversing TBP-induced LTP. (E) LFS given outside this reversal window does not affect spine size. (F) LFS given outside the reversal window does not reverse LTP either. Taken from [34, 71].

and Linden demonstrated that LTD expression and spine changes in cerebellar Purkinje cells could be induced independently of each other, and induction of one did not affect the other [92]. Wang et al. reported that trafficking of AMPARs to and away from PSDs was activityindependent and not associated with alterations in spine size. The significance of this finding requires further investigation [70].

One interesting and important feature of synaptic modification is its reversibility. This reversibility is defined by reversal of synaptic modification after its induction [34, 93]. There are a few aspects to this reversal: (1) reversal applies to both LTP and LTD, and spine enlargement and spine shrinkage [34, 71] (**Figure 4**). More specifically, low frequency stimulation reverses LTP and spine enlargement, while high frequency stimulation reverses LTD and spine shrinkage. (2) There is a critical time window only during which reversal can occur [34, 71, 93, 94] (**Figure 4**). In hippocampal slices, the window for LTP reversal is about 15–30 min [71]. (3) The typical stimuli that can induce reversal are not capable altering basal synaptic strength or spine dimension [71, 93, 95, 96].

then move laterally into spines/PSDs. Thus, with LTP induction, two rapid processes (within 30 sec) occurs independent of each other: spine expansion which requires actin polymerization and activation of NMDARs but not postsynaptic PKA activation, and delivery of AMPARs to the perisynaptic regions which requires activation of NMDARs and postsynaptic PKA signaling but not actin polymerization. The next 15 min or so (reversal time window) determines whether LTP and spine expansion can be stabilized into a long-term change. During this period, translocation of the newly delivered perisynaptic AMPARs stabilized spine expansion, while removal of these receptors led to collapse of enlarged spines [71]. On the other hand, reversal of spine enlargement also removed these perisynaptic AMPARs. Hence, there is a mutual interaction between perisynaptic AMPARs and spine enlargement in that the presence of one is required to sustain the other (**Figure 3**). Yang et al. found that postsynaptic PKC activity is required for the translocation of perisynaptic AMPARs to synapse, and in the absence of PKC activity, these AMPARs remain perisynaptic. Importantly, as long as the perisynaptic AMPARs are present, both LTP and spine expansion exist in a labile state in that they can either be reverted to the baseline state (no plasticity), or they can enter a stabilized state of persistent increase in synaptic strength and spine size (persistent plasticity). Low frequency synaptic stimulation given within a 15 min "grace period" post-LTP induction reversed spine expansion and removed AMPARs from the perisynaptic regions and hence blocked the conversion of short-term plasticity to a long-

Dendritic Spine Modifications in Brain Physiology http://dx.doi.org/10.5772/intechopen.73326 157

**Figure 5.** Two-step model for coordinated expression of synaptic potentiation and spine enlargement. TBP triggers two initial processes, spine enlargement and perisynaptic delivery of AMPARs. These two processes occur simultaneously and mostly independent of each other. In the absence of any disturbance, perisynaptic AMPARs translocate into synapse, which stabilizes both AMPARs and spine enlargement, and this leads to persistent potentiation of synaptic responses and spine enlargement. On the other hand, when low frequency synaptic stimulation (LFS) is given within this "grace period" when both processes are in a liable state, it readily removes perisynaptic AMPARs. As a result, synaptic potentiation is aborted and spine enlargement collapses, and no LTP and spine enlargement. This model shows the interaction between functional and structural aspects of synapse modification is critical to the persistency and long-

term one (**Figure 5**) [71, 82].

lasting occurrence of synaptic modification.

Although it is generally believed that expression of LTP requires the addition of synaptic AMPARs, some evidences suggest that these newly added AMPARs are not delivered directly into the PSDs inside spines, but rather they are either delivered to regions outside synapses (i.e., the perisynaptic regions; [82] or onto dendritic shaft [83]. These AMPARs

**Figure 4.** Actin polymerization is required for spine expansion and maintenance of perisynaptic AMPA receptors. (A) Actin depolymerizing agent Latrunculin A (LatA) added before and during TBP abolished spine expansion. (B) Bath perfusion of LatA before and during TBP impaired enhancement of synaptic response (EPSPs). (C) Bath perfusion of LatA 2 min after TBP did not disrupt the delivery of AMPARs to perisynaptic site, as revealed by the increased in responses to application of glutamate transporter blocker TBOA (open symbols). (D) Bath perfusion of LatA 2 min after TBP did not affect spine enlargement, indicating that persistent spine enlargement does not require persistent actin polymerization. (E) Bath perfusion of LatA 2 min after TBP prevented the occurrence of gradual increase in EPSP enhancement which requires the synaptic addition of new AMPARs. (F) Addition of LatA 2 min after TBP removed the newly delivered perisynaptic AMPARs as revealed by the absence of increase in response to TBOA. Taken from [83].

then move laterally into spines/PSDs. Thus, with LTP induction, two rapid processes (within 30 sec) occurs independent of each other: spine expansion which requires actin polymerization and activation of NMDARs but not postsynaptic PKA activation, and delivery of AMPARs to the perisynaptic regions which requires activation of NMDARs and postsynaptic PKA signaling but not actin polymerization. The next 15 min or so (reversal time window) determines whether LTP and spine expansion can be stabilized into a long-term change. During this period, translocation of the newly delivered perisynaptic AMPARs stabilized spine expansion, while removal of these receptors led to collapse of enlarged spines [71]. On the other hand, reversal of spine enlargement also removed these perisynaptic AMPARs. Hence, there is a mutual interaction between perisynaptic AMPARs and spine enlargement in that the presence of one is required to sustain the other (**Figure 3**). Yang et al. found that postsynaptic PKC activity is required for the translocation of perisynaptic AMPARs to synapse, and in the absence of PKC activity, these AMPARs remain perisynaptic. Importantly, as long as the perisynaptic AMPARs are present, both LTP and spine expansion exist in a labile state in that they can either be reverted to the baseline state (no plasticity), or they can enter a stabilized state of persistent increase in synaptic strength and spine size (persistent plasticity). Low frequency synaptic stimulation given within a 15 min "grace period" post-LTP induction reversed spine expansion and removed AMPARs from the perisynaptic regions and hence blocked the conversion of short-term plasticity to a longterm one (**Figure 5**) [71, 82].

and spine shrinkage. (2) There is a critical time window only during which reversal can occur [34, 71, 93, 94] (**Figure 4**). In hippocampal slices, the window for LTP reversal is about 15–30 min [71]. (3) The typical stimuli that can induce reversal are not capable altering basal

Although it is generally believed that expression of LTP requires the addition of synaptic AMPARs, some evidences suggest that these newly added AMPARs are not delivered directly into the PSDs inside spines, but rather they are either delivered to regions outside synapses (i.e., the perisynaptic regions; [82] or onto dendritic shaft [83]. These AMPARs

**Figure 4.** Actin polymerization is required for spine expansion and maintenance of perisynaptic AMPA receptors. (A) Actin depolymerizing agent Latrunculin A (LatA) added before and during TBP abolished spine expansion. (B) Bath perfusion of LatA before and during TBP impaired enhancement of synaptic response (EPSPs). (C) Bath perfusion of LatA 2 min after TBP did not disrupt the delivery of AMPARs to perisynaptic site, as revealed by the increased in responses to application of glutamate transporter blocker TBOA (open symbols). (D) Bath perfusion of LatA 2 min after TBP did not affect spine enlargement, indicating that persistent spine enlargement does not require persistent actin polymerization. (E) Bath perfusion of LatA 2 min after TBP prevented the occurrence of gradual increase in EPSP enhancement which requires the synaptic addition of new AMPARs. (F) Addition of LatA 2 min after TBP removed the newly delivered perisynaptic AMPARs as revealed by the absence of increase in response to TBOA. Taken from [83].

synaptic strength or spine dimension [71, 93, 95, 96].

156 Neuroplasticity - Insights of Neural Reorganization

**Figure 5.** Two-step model for coordinated expression of synaptic potentiation and spine enlargement. TBP triggers two initial processes, spine enlargement and perisynaptic delivery of AMPARs. These two processes occur simultaneously and mostly independent of each other. In the absence of any disturbance, perisynaptic AMPARs translocate into synapse, which stabilizes both AMPARs and spine enlargement, and this leads to persistent potentiation of synaptic responses and spine enlargement. On the other hand, when low frequency synaptic stimulation (LFS) is given within this "grace period" when both processes are in a liable state, it readily removes perisynaptic AMPARs. As a result, synaptic potentiation is aborted and spine enlargement collapses, and no LTP and spine enlargement. This model shows the interaction between functional and structural aspects of synapse modification is critical to the persistency and longlasting occurrence of synaptic modification.

The above results are consist with a model (**Figure 5**) in which (1) the expression of functional plasticity (LTP) and structural plasticity (spine enlargement) are initially two independent processes only share the activation of NMDARs; (2) these two processes then enter an interactive state that the continuous presence of one is required for the persistence of the other; (3) the above state is liable in that interference (such as low frequency stimulation) can revert both changes back to the baseline; (4) once both processes are stabilized, synapse modification has entered a state resistant to reversal. We like to note that during the "grace period" coordinated changes in synaptic plasticity (function and structure) are cross-checked to ensure that they do occur together, and in the situation only one such process occurs (perhaps can be viewed as a mistake), the other process will be aborted albeit in process. This double-proof mechanism is essential to ensure that only appropriate changes are allowed to be sustained, and may thus be especially important in face of the highly dynamic nature of synaptic modifications, such as those occurring during early neural development [94].

and structure. Extensive progress has been made on our understanding of the structure and function of spine which vastly has advanced our understanding of neuronal and synaptic communication and plasticity. In addition, changes in spine density and dimension may serve as a marker of pathological processes and hence have potential therapeutic/diagnostic

Dendritic Spine Modifications in Brain Physiology http://dx.doi.org/10.5772/intechopen.73326 159

This work was supported by grants from Peking University Shenzhen Graduate School and grants from Shenzhen government (ZDSYS201504301539161 and KQTD2015032709315529).

School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate

[1] Tackenberg C, Ghori A, Brandt R. Thin, stubby or mushroom: Spine pathology in

[2] Peters A, Kaiserman-Abramof IR. The small pyramidal neuron of the rat cerebral cortex. The perikaryon, dendrites and spines. The American Journal of Anatomy. 1970;**127**

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values.

**Acknowledgements**

**Competing interests**

**Author details**

Jun Ju and Qiang Zhou\*

School, Shenzhen, China

(4):321-355

Neuroscience. 1992;**12**(7):2685-2705

Neurobiology. 1985;**5**(3):271-284

**References**

The authors declare that they have no competing interests.

\*Address all correspondence to: zhouqiang@pkusz.edu.cn

Sustained reduction in synaptic strength may eventually lead to the loss of synaptic connections, and this loss is manifested as a reduction in spine density. Spine loss appears to be a protracted process and hence it is difficult to study. Even if monitoring changes in the same set of synapses/spines, it is usually more difficult to exclude the possibility that the reduced synaptic function and spine number is caused by deterioration of the health of the preparation, or by some other unknown or uncontrolled processes that occur randomly during the long period (>hours) between LTD induction and spine loss. Nonetheless, a few studies have examined this process. By using organotypical slices and monitoring both presynaptic boutons and spines, Becker et al. showed that LTD induction increased the turnover rate of presynaptic boutons and resulted in decreased synaptic contacts between the pre- and post-synaptic sites. Although presynaptic boutons and postsynaptic spines disappear at much greater rate after LTD, there is no particular pattern to follow, since disappearance of either presynaptic boutons or spines could occur prior to the other [97]. Therefore, the above observations suggest that the mismatch between presynaptic and postsynaptic sites is more likely a key factor in the elimination of synapse, while the exact sequence might not play much role.
