Changes In Dendritic Structure Research Articles

PGC-1α in the hippocampus mediates depressive-like and stress-coping behaviours and regulates excitatory synapses in the dentate gyrus in mice

Decreased hippocampal synaptic plasticity is an important pathological change in stress-related mood disorders, including major depressive disorder. However, the underlying mechanism is unclear. PGC-1α, a transcriptional coactivator, is a key factor in synaptic plasticity. We investigated the relationships between changes in hippocampal PGC-1α expression and depressive-like and stress-coping behaviours, and whether they are related to hippocampal synapses. Adeno-associated virus was used to alter hippocampal PGC-1α expression in male C57BL/6 mice. The sucrose preference test and forced swimming test were used to assess their depressive-like and stress-coping behaviours, respectively. Immunohistochemistry and stereology were used to calculate the total number of excitatory synapses in each hippocampal subregion (the cornu ammonis (CA) 1, CA3, and dentate gyrus). Immunofluorescence was used to visualize the changes in dendritic structure. Western blotting was used to detect the expression of hippocampal PGC-1α and mitochondrial-associated proteins, such as UCP2, NRF1 and mtTFAs. Our results showed that mice with downregulated PGC-1α expression in the hippocampus exhibited depressive-like and passive stress-coping behaviours, while mice with upregulated PGC-1α in the hippocampus exhibited increased stress-coping behaviours. Moreover, the downregulation of hippocampal PGC-1α expression resulted in a decrease in the number of excitatory synapses in the DG and in the protein expression of UCP2 in the hippocampus. Alternatively, upregulation of hippocampal PGC-1α yielded the opposite results. This suggests that hippocampal PGC-1α is involved in regulating depressive-like and stress-coping behaviours and modulating the number of excitatory synapses in the DG. This provides new insight for the development of antidepressants.

Abstract WP293: Elucidating the Neuronal Basis of Diaschisis in Stroke Recovery

Numerous studies have shown that focal ischemic stroke changes cortical activity patterns, metabolism and blood flow not only in peri-infarct regions, but also in remote regions connected to the ischemic core. This phenomenon known as “diaschesis”, was predicted over 100 years ago and is presumed to play an important role in dictating the extent of stroke recovery. Although regional brain imaging studies provide some support to this idea (with caveats), direct examination of the structure and function of neurons connected to neurons lost in the infarct core, has not been adequately addressed. One reason for our lack of a detailed understanding is the methodologies to trace connected neurons and track their function/activity patterns over weeks and months time after stroke, did not exist until recently. To address this issue, we injected adeno-associated virus (AAV) into the adult mouse primary forelimb somatosensory cortex (FLS1) 2-3 weeks before induction of photothrombotic stroke. These AAVs could either be transported in a retrograde fashion to label neural inputs to the FLS1 (retro pAAV.CAG.GFP), or conversely move in an anterograde, trans-synaptic manner (AAV.Syn.Cre) to label neurons downstream of FLS1 neurons, such as in motor cortex. In the latter case, we utilized a transgenic mouse that co-expressed a cre-recombinase dependent fluorescent reporter (tdTomato), as well as a genetically encoded calcium sensor (GCaMP6s) in neurons. These dual reporters (tdTomato and GCaMP6s) allowed us to examine changes in dendritic structure as well as functional activity patterns in downstream motor neurons. Our initial findings using confocal microscopy to assess dendritic structure, indicate subtle changes in spine density in neuronal inputs and outputs of the FLS1, within the first week after stroke. Repeated in vivo imaging of calcium transients in downstream motor cortex neurons in awake mice before and after FLS1 cortex stroke, suggested a long lasting disruption in neuronal and ensemble activity patterns. These preliminary findings provide direct evidence that neurons in remotely connected brain regions suffer long-lasting impairments after stroke, thereby providing another potential target or substrate for future therapeutics to consider.

Sleep and memory: The impact of sleep deprivation on transcription, translational control, and protein synthesis in the brain.

In countries around the world, sleep deprivation represents a widespread problem affecting school-age children, teenagers, and adults. Acute sleep deprivation and more chronic sleep restriction adversely affect individual health, impairing memory and cognitive performance as well as increasing the risk and progression of numerous diseases. In mammals, the hippocampus and hippocampus-dependent memory are vulnerable to the effects of acute sleep deprivation. Sleep deprivation induces changes in molecular signaling, gene expression and may cause changes in dendritic structure in neurons. Genome wide studies have shown that acute sleep deprivation alters gene transcription, although the pool of genes affected varies between brain regions. More recently, advances in research have drawn attention to differences in gene regulation between the level of the transcriptome compared with the pool of mRNA associated with ribosomes for protein translation following sleep deprivation. Thus, in addition to transcriptional changes, sleep deprivation also affects downstream processes to alter protein translation. In this review, we focus on the multiple levels through which acute sleep deprivation impacts gene regulation, highlighting potential post-transcriptional and translational processes that may be affected by sleep deprivation. Understanding the multiple levels of gene regulation impacted by sleep deprivation is essential for future development of therapeutics that may mitigate the effects of sleep loss.

Imipramine Ameliorates Depressive Symptoms by Blocking Differential Alteration of Dendritic Spine Structure in Amygdala and Prefrontal Cortex of Chronic Stress-Induced Mice.

Previous studies have shown disrupted synaptic plasticity and neural activity in depression. Such alteration is strongly associated with disrupted synaptic structures. Chronic stress has been known to induce changes in dendritic structure in the basolateral amygdala (BLA) and medial prefrontal cortex (mPFC), but antidepressant effect on structure of these brain areas has been unclear. Here, the effects of imipramine on dendritic spine density and morphology in BLA and mPFC subregions of stressed mice were examined. Chronic restraint stress caused depressive-like behaviors such as enhanced social avoidance and despair level coincident with differential changes in dendritic spine structure. Chronic stress enhanced dendritic spine density in the lateral nucleus of BLA with no significant change in the basal nucleus of BLA, and altered the proportion of stubby or mushroom spines in both subregions. Conversely, in the apical and basal mPFC, chronic stress caused a significant reduction in spine density. The proportion of stubby or mushroom spines in these subregions overall reduced while the proportion of thin spines increased after repeated stress. Interestingly, most of these structural alterations by chronic stress were reversed by imipramine. In addition, structural changes caused by stress and blocking the changes by imipramine were corelated well with altered activation and expression of synaptic plasticity-promoting molecules such as phospho-CREB, phospho-CAMKII, and PSD-95. Collectively, our data suggest that imipramine modulates stress-induced changes in synaptic structure and synaptic plasticity-promoting molecules in a coordinated manner although structural and molecular alterations induced by stress are distinct in the BLA and mPFC.

The SEM Metallography Analysis of Vacuum Cast ZhS6K Superalloy Turbine Blade after Various Working Hours

A vacuum cast Ni-base superalloy ZhS6K is used for the production of turbine blades whose are working in the hot section of turbine in former USSR DV-2 jet engine. The usual working temperature for this kind of alloy is designed for 800/1050°C. Turbine blades with protective alitized layer made from this alloy were evaluated at the starting stage, then after 600, 1000, 1500 and 2000 hours of regular working. The SEM methods of quantitative metallography were used for structural characteristics evaluation. The microstructural analysis shows increasing of the medium distance of secondary dendrite arms D, what is related to chemical changes in dendritic structure, and the intermetallic strengthening gamma prime phase coarse and turn into spherical shape what leads to decreasing of dislocation and precipitation strengthening mechanism and further decreasing of hot temperature properties of the alloy. The method used at quantitative evaluation and techniques of colour optical microscopy and SEM is useful when microstructure degradation of superalloys has occurred.

Motor Areas Show Altered Dendritic Structure in an Amyotrophic Lateral Sclerosis Mouse Model.

Objective: Motor neurons (MNs) die in amyotrophic lateral sclerosis (ALS), a clinically heterogeneous neurodegenerative disease of unknown etiology. In human or rodent studies, MN loss is preceded by increased excitability. As increased neuronal excitability correlates with structural changes in dendritic arbors and spines, we have examined longitudinal changes in dendritic structure in vulnerable neuron populations in a mouse model of familial ALS.Methods: We used a modified Golgi-Cox staining method to determine the progressive changes in dendritic structure of hippocampal CA1 pyramidal neurons, striatal medium spiny neurons, and resistant (trochlear, IV) or susceptible (hypoglossal, XII; lumbar) MNs from brainstem and spinal cord of mice over-expressing the human SOD1G93A (SOD1) mutation, in comparison to wild-type (WT) mice, at four postnatal (P) ages of 8–15, 28–35, 65–75, and 120 days.Results: In SOD1 mice, dendritic changes occur at pre-symptomatic ages in both XII and spinal cord lumbar MNs. Spine loss without dendritic changes was present in striatal neurons from disease onset. Spine density increases were present at all ages studied in SOD1 XII MNs. Spine density increased in neonatal lumbar MNs, before decreasing to control levels by P28-35 and was decreased by P120. SOD1 XII MNs and lumbar MNs, but not trochlear MNs showed vacuolization from the same time-points. Trochlear MN dendrites were unchanged.Interpretation: Dendritic structure and spine alterations correlate with the neuro-motor phenotype in ALS and with cognitive and extra-motor symptoms seen in patients. Prominent early changes in dendritic arbors and spines occur in susceptible cranial and spinal cord MNs, but are absent in MNs resistant to loss in ALS.

Marked changes in dendritic structure and spine density precede significant neuronal death in vulnerable cortical pyramidal neuron populations in the SOD1(G93A) mouse model of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is characterised by the death of upper (corticospinal) and lower motor neurons (MNs) with progressive muscle weakness. This incurable disease is clinically heterogeneous and its aetiology remains unknown. Increased excitability of corticospinal MNs has been observed prior to symptoms in human and rodent studies. Increased excitability has been correlated with structural changes in neuronal dendritic arbors and spines for decades. Here, using a modified Golgi-Cox staining method, we have made the first longitudinal study examining the dendrites of pyramidal neurons from the motor cortex, medial pre-frontal cortex, somatosensory cortex and entorhinal cortex of hSOD1G93A (SOD1) mice compared to wild-type (WT) littermate controls at postnatal (P) days 8–15, 28–35, 65–75 and 120. Progressive decreases in dendritic length and spine density commencing at pre-symptomatic ages (P8-15 or P28-35) were observed in layer V pyramidal neurons within the motor cortex and medial pre-frontal cortex of SOD1 mice compared to WT mice. Spine loss without concurrent dendritic pathology was present in the pyramidal neurons of the somatosensory cortex from disease-onset (P65-75). Our results from the SOD1 model suggest that dendritic and dendritic spine changes foreshadow and underpin the neuromotor phenotypes present in ALS and may contribute to the varied cognitive, executive function and extra-motor symptoms commonly seen in ALS patients. Determining if these phenomena are compensatory or maladaptive may help explain differential susceptibility of neurons to degeneration in ALS.

Characteristic patterns of dendritic remodeling in early-stage glaucoma: evidence from genetically identified retinal ganglion cell types.

Retinal ganglion cell (RGC) loss is a hallmark of glaucoma and the second leading cause of blindness worldwide. The type and timing of cellular changes leading to RGC loss in glaucoma remain incompletely understood, including whether specific RGC subtypes are preferentially impacted at early stages of this disease. Here we applied the microbead occlusion model of glaucoma to different transgenic mouse lines, each expressing green fluorescent protein in 1-2 specific RGC subtypes. Targeted filling, reconstruction, and subsequent comparison of the genetically identified RGCs in control and bead-injected eyes revealed that some subtypes undergo significant dendritic rearrangements as early as 7 d following induction of elevated intraocular pressure (IOP). By comparing specific On-type, On-Off-type and Off-type RGCs, we found that RGCs that target the majority of their dendritic arbors to the scleral half or "Off" sublamina of the inner plexiform layer (IPL) undergo the greatest changes, whereas RGCs with the majority of their dendrites in the On sublamina did not alter their structure at this time point. Moreover, M1 intrinsically photosensitive RGCs, which functionally are On RGCs but structurally stratify their dendrites in the Off sublamina of the IPL, also underwent significant changes in dendritic structure 1 week after elevated IOP. Thus, our findings reveal that certain RGC subtypes manifest significant changes in dendritic structure after very brief exposure to elevated IOP. The observation that RGCs stratifying most of their dendrites in the Off sublamina are first to alter their structure may inform the development of new strategies to detect, monitor, and treat glaucoma in humans.

Automated and quantitative image analysis of ischemic dendritic blebbing using in vivo 2-photon microscopy data

Ischemia induces a ‘blebbing’ of dendrites, a structural alteration where dendrites take on a ‘beads on a string’ appearance. We developed a toolkit program, BlebQuant, for quantitative automated bleb analysis to chart the morphology of dendrites labeled with GFP/YFP under normal conditions and after ischemia-induced damage. In vivo 2-photon data from mouse layer 5 neurons with apical dendritic tufts extending to the cortical surface were examined before, during, and after global ischemia. To quantify changes in dendritic structure, we used morphometric tools that exploit characteristic features of blebbing, distinguished as localized regions of spherical or ellipsoid swellings. By comparing acquired images during ischemia and reperfusion to a pre-ischemia reference image, our automated approach detected blebs based on defined eccentricity and area thresholds and quantified the percentage of blebbed dendrites based on a block-selection method. Our results indicate that the automated morphometric indices we employ yield results that correlate with manual assessment. The automated approach permits rapid and effective analysis of dendritic structure and may facilitate the study of ischemic dendritic damage.

Changes in dendritic morphology and spine density in motor cortex of the adult rat after stroke during infancy

Cognitive and motor deficits are pervasive in children that suffer early brain injury. The aim of this study was to determine the impact that early damage has on dendritic spine density and other aspects of dendritic morphology of neurons in the motor cortex. Also of interest was how changes in dendritic structure evolved across the lifespan. Ischemia was induced in 10-day-old Long Evans rats by injection of Rose Bengal dye and a laser positioned over right motor cortex. Animals were sacrificed at two and six months of age, and brains were processed for Golgi-Cox staining. Animals exposed to early damage exhibited increases in length of basilar dendrites at two months of age, however no differences in spine density were found across groups at this age. At six months of age, injured animals demonstrated an overall decrease in apical and basilar spine density. Our results suggest that the changes in dendritic length and spine density observed after early damage are unable to be maintained as the animal ages. The observation that increases in spine density do not necessarily coincide with increases in dendritic length suggests that the two processes may not be dependent on one another and suggest two independent plasticity processes responding to damage.

Reversal of long-term dendritic spine alterations in Alzheimer disease models

Synapse loss is strongly correlated with cognitive impairment in Alzheimer's disease (AD). We have previously reported the loss of dendritic spines and the presence of dystrophic neurites in both the hippocampi of transgenic mice overexpressing amyloid precursor protein (APP) and in the human brain affected with AD. In the studies reported here we have asked whether the acute alterations in dendritic spines induced by Abeta, as well as the chronic loss of spine density seen in hAPP transgenic mice, are reversible by treatments that restore the cAMP/PKA/CREB signaling pathway or proteasome function to control levels. The results show that both rolipram and TAT-HA-Uch-L1 restore spine density to near control conditions, even in elderly mice. The results suggest that changes in dendritic structure and function that occur after Abeta elevation are reversible even after long periods of time, and that one could envision therapeutic approaches to AD based on this restoration that could work independently of therapies aimed at lowering Abeta levels in the brain.

Prion Protein with an Insertional Mutation Accumulates on Axonal and Dendritic Plasmalemma and Is Associated with Distinctive Ultrastructural Changes

Prion diseases are fatal neurological diseases characterized by central nervous system deposition of abnormal forms of a membrane glycoprotein designated PrP (prion protein). Tg(PG14) transgenic mice express PrP that harbor a nine-octapeptide insertional mutation homologous to one described in a familial prion disease of humans. Tg(PG14) mice spontaneously develop a fatal neurological illness accompanied by massive apoptosis of cerebellar granule neurons and accumulation of an aggregated and weakly protease-resistant form of PrP that is not infectious. Previous light microscopic analyses of these mice left open questions regarding the subcellular distribution of the mutant protein and the nature of the neuropathological lesions produced. To address these questions, we undertook an immunogold electron microscopic study of Tg(PG14) mice. We found that mutant PrP is localized primarily on the plasma membrane of dendrites and unmyelinated axons in the hippocampus and cerebellum, with little labeling of either neuronal cell bodies or intracellular organelles. PrP deposits were shown to be associated with degenerative changes in dendritic structure. We also describe for the first time marked pathology in myelinated axons, and alterations in the axon/oligodendrocyte interface. Taken together, our results suggest cellular mechanisms by which mutant PrPs produce pathology. In addition, they highlight distinctions between familial and infectious prion disorders at the ultrastructural level that correlate with differences in cellular trafficking of the disease-associated PrP forms.

Two-Photon Imaging of Stroke OnsetIn VivoReveals That NMDA-Receptor Independent Ischemic Depolarization Is the Major Cause of Rapid Reversible Damage to Dendrites and Spines

We adapt a mouse global ischemia model to permit rapid induction of ischemia and reperfusion in conjunction with two-photon imaging to monitor the initial ionic, structural, and functional implications of brief interruptions of blood flow (6-8 min) in vivo. After only 2-3 min of global ischemia, a wide spread loss of mouse somatosensory cortex apical dendritic structure is initiated during the passage of a propagating wave (3.3 mm/min) of ischemic depolarization. Increases in intracellular calcium levels occurred during the wave of ischemic depolarization and were coincident with the loss of dendritic structure, but were not triggered by reperfusion. To assess the role of NMDA receptors, we locally applied the antagonist MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate] at concentrations sufficient to fully block local NMDA agonist-evoked changes in intracellular calcium levels in vivo. Changes in dendritic structure and intracellular calcium levels were independent of NMDA receptor activation. Local application of the non-NMDA glutamate receptor antagonist CNQX also failed to block ischemic depolarization or rapid changes in dendrite structure. Within 3-5 min of reperfusion, damage ceased and restoration of synaptic structure occurred over 10-60 min. In contrast to a reperfusion promoting damage, over this time scale, the majority of spines and dendrites regained their original structure during reperfusion. Intrinsic optical signal imaging of sensory evoked maps indicated that reversible alteration in dendritic structure during reperfusion was accompanied by restored functional maps. Our results identify glutamate receptor-independent ischemic depolarization as the major ionic event associated with disruption of synaptic structure during the first few minutes of ischemia in vivo.

Chronic phencyclidine treatment increases dendritic spine density in prefrontal cortex and nucleus accumbens neurons

We examined whether repeated exposure to the noncompetitive NMDA receptor antagonist phencyclidine (PCP) produces enduring changes in dendritic structure in a manner similar to the stimulants cocaine and amphetamine. Adult rats were treated with i.p. injections of PCP (5 mg/kg) or saline, twice a day, for 5 consecutive days, for a total of 4 weeks. One month after the last injection, their brains were removed and processed for Golgi-Cox staining. Prior exposure to PCP increased dendritic spine density in the mPFC and NAcc core, but not in the parietal cortex. These findings, which are similar to those observed after chronic treatment with cocaine and amphetamine, raise the possibility that, despite differences in their mechanisms of action, PCP and stimulant drugs may induce some of their enduring effects via common processes.

Hepatocyte growth factor‐induced enhancement of dendritic branching is blocked by inhibitors of N‐methyl‐D‐aspartate receptors and calcium/calmodulin‐dependent kinases

Hepatocyte growth factor (HGF) and its receptor, Met, are clustered at excitatory synapses and can enhance N-methyl-D-aspartate (NMDA) receptor current and promote formation of neurites and dendrites. In this study, we examine the effects of HGF on dendritic arborization in mature cultures of dissociated hippocampal neurons. Exogenous HGF treatment caused a dose-dependent increase in total dendritic branch tip number, total dendritic branch length, and dendritic complexity in these neurons. NMDA receptor activity has been linked to changes in dendritic structure, so we tested the effects of HGF on the dendritic arbor in the presence of DL-2-amino-5-phosphonopentanoic acid (APV), an NMDA receptor inhibitor. APV blocked the HGF-induced enhancement of the dendritic arbor in a dose-dependent manner. Similarly, pretreatment of neurons with KN62, an inhibitor of calcium-dependent kinases, suppressed changes in dendritic branching induced by HGF. These results suggest that HGF initiates Ca2+-dependent processes, so we examined the effect of HGF on intracellular calcium levels and autophosphorylation of the calcium/calmodulin-dependent protein kinase II (CaMKII). HGF caused a persistent increase in fluorescence in clusters along dendrites of neurons preloaded with the Ca2+ indicator Fluo-4. HGF treatment also enhanced autophosphorylation of CaMKII. The increases in Fluo-4 fluorescence and autophosphorylation of CaMKII were blocked by pretreatment of neurons with APV. These results indicate that HGF stimulates Ca2+ influx into dendrites through the NMDA receptor and that this effect is necessary for the changes in dendritic morphology induced by HGF.

Methamphetamine‐induced structural plasticity in the dorsal striatum

Repeated exposure to psychostimulant drugs produces long-lasting changes in dendritic structure, presumably reflecting a reorganization in patterns of synaptic connectivity, in brain regions that mediate the psychomotor activating and incentive motivational effects of these drugs, including the nucleus accumbens and prefrontal cortex. However, repeated exposure to psychostimulant drugs also facilitates a transition in the control of some behaviors from action-outcome associations to behavior controlled by stimulus-response (S-R) habits. This latter effect is thought to be due to increasing engagement and control over behavior by the dorsolateral (but not dorsomedial) striatum. We hypothesized therefore that repeated exposure to methamphetamine would differentially alter the density of dendritic spines on medium spiny neurons (MSNs) in the dorsolateral vs. dorsomedial striatum. Rats were treated with repeated injections of methamphetamine, and 3 months later dendrites were visualized using Sindbis virus-mediated green fluorescent protein (GFP) expression in vivo. We report that prior exposure to methamphetamine produced a significant increase in mushroom and thin spines on MSNs in the dorsolateral striatum, but a significant decrease in mushroom spines in the dorsomedial striatum. This may be due to changes in the glutamatergic innervation of these two subregions of the dorsal striatum. Thus, we speculate that exposure to psychostimulant drugs may facilitate the development of S-R habits because this reorganizes patterns of synaptic connectivity in the dorsal striatum in a way that increases control over behavior by the dorsolateral striatum.

Exercise-induced changes in dendritic structure and complexity in the adult hippocampal dentate gyrus

Neurogenesis is a constitutive activity in the adult dentate gyrus whereby new cells are created in the subgranular zone, before becoming neurons in the dentate gyrus granule cell layer. New granule cells are thought to migrate from the subgranular zone outwards to the edge of the cell layer as they mature. In these experiments we examined the dendritic morphology of granule cells in the subgranular zone, and the inner and outer regions of the granule cell zone in Sprague–Dawley rats with low and high rates of neurogenesis. In animals with lower rates of neurogenesis, the number of primary dendrites, degree of dendritic complexity and total dendritic length was lowest in cells located in the subgranular zone, higher in inner granule cell zone neurons, and highest in outer granule cell zone granule cells. Subgranular zone granule cells typically extended one primary dendrite and had a simple, immature dendritic tree, while granule cells in the outer granule cell zone had an increased number of primary dendrites, greater dendritic complexity, and greater total dendritic length. Animals that engaged in voluntary exercise showed increased neurogenesis, and the proportion of cells with one or two primary dendrites was increased in all of the granule cell zones. Despite having fewer primary processes, these cells showed enhanced dendritic complexity and an overall increase in their total dendritic length. These results indicate that granule cell dendritic morphology may be indicative of the age and position of a cell in the granule cell layer, but that in animals with increased rates of neurogenesis, the proportion of cells exhibiting what is considered an immature phenotype is increased throughout the all regions of the dentate gyrus cell layer.

Large-scale changes in dendritic structure during reorganization of adult somatosensory cortex

In adult rat somatosensory cortex (S1), neurons are biased and have less dendritic arbor close to the border between the forepaw and lower jaw representations. Changes in sensory experience cause changes in the functional organization of the neocortex. Therefore, we examined the morphology of neurons in the reorganized region of S1 after forepaw denervation. We found that during reorganization dendritic arbors changed to reflect the new location of the border.

Synaptic Remodeling through MAPK

Synaptic plasticity is a model for learning and memory. Dendrites, the sites of excitatory synaptic contacts, undergo morphological changes in response to stimuli. Wu et al. investigated the role of the mitogen-activated protein kinase (MAPK) pathway in changes in dendritic structure in response to multiple brief depolarizations over a period of minutes in cultured neurons or explants. Single stimuli (depolarizations induced by elevated extracellular K + ) of long or short duration produced a transient activation of the MAPKs, ERK1 and ERK2, measured as phosphorylation of these MAPKs. However, repeated spaced depolarizations produced a prolonged MAPK phosphorylation. Application of pharmacological inhibitors showed that the initial increase in MAPK activity depended on an increase in the concentration of free intracellular Ca 2+ , activation of calmodulin and calcium/calmodulin-dependent kinase, the small guanosine triphosphatase Ras, and MEK (the upstream kinase for ERK1 and 2). Persistent MAPK phosphorylation required continuous activity of MEK and was dependent on protein kinase A. In explants of dentate gyrus transfected to express green fluorescent protein, MAPK activity was found to be essential for changes in dendrite morphology (filopodial extensions and the formation of new spines) produced by multiple spaced stimulations. G.-Y. Wu, K. Deisseroth, R.W. Tsien, Spaced stimuli stabilize MAPK pathway activation and its effects on dendritic morphology. Nature Neurosci. 4 , 151-158 (2001). [Online Journal]

Developmental switch in excitability, Ca(2+) and K(+) currents of retinal ganglion cells and their dendritic structure.

In the retina of teleost fish, continuous neuronal development occurs at the margin, in the peripheral growth zone (PGZ). We prepared tissue slices from the retina of rainbow trout that include the PGZ and that comprise a time line of retinal development, in which cells at progressive stages of differentiation are present side by side. We studied the changes in dendritic structure and voltage-dependent Ca(2+), Na(+), and K(+) currents that occur as ganglion cells mature. The youngest ganglion cells form a distinct bulge. Cells in the bulge have spare and short dendritic trees. Only half express Ca(2+) currents and then only high-voltage-activated currents with slow inactivation (HVAslow). Bulge cells are rarely electrically excitable. They express a mixture of rapidly inactivating and noninactivating K(+) currents (IKA and IKdr). The ganglion cells next organize into a transition zone, consisting of a layered structure two to three nuclei thick, before forming the single layered structure characteristic of the mature retina. In the transition zone, the dendritic arbor is elaborately branched and extends over multiple laminae in the inner plexiform layer, without apparent stratification. The arbor of the mature cells is stratified, and the span of the dendritic arbor is well over five times the cell body's diameter. The electrical properties of cells in the transition and mature zones differ significantly from those in the bulge cells. Correlated with the more elaborate dendritic structures are the expression of both rapidly inactivating HVA (HVAfast) and of low-voltage-activated (LVA) Ca(2+) currents and of a high density of Na(+) currents that renders the cells electrically excitable. The older ganglion cells also express a slowly activating K(+) current (IKsa).