Activity-dependent Potentiation Research Articles

Potentiation of force by extracellular potassium is not dependent on muscle length in mouse EDL muscle.

Increases in myofiber extracellular potassium with prolonged contractile activity can potentiate twitch force. Activity-dependent potentiation, another mechanism of force increase in skeletal muscle, has a strong dependence on muscle or sarcomere length. Thus, potassium-mediated twitch potentiation could also be length-dependent. However, this has not been previously investigated. To this end, we used isolated C57BL/6 mouse extensor digitorum longus (EDL) muscles and elicited twitches at 0.9 Lo, Lo, and 1.1 Lo (Lo refers to optimal length) in normal (5 mM) and high (10 mM) potassium solutions. Potentiation magnitude was similar to previous observations and was not significantly different between lengths (0.9 Lo: 12.3 ± 4.4%, Lo: 12.2 ± 3.6%, 1.1 Lo: 11.8 ± 4.8%, values are means ± SD). Exposure to dantrolene sodium, a compound that attenuates calcium release, reduced twitch force across lengths by ∼70%. When dantrolene-affected muscles were subsequently exposed to high potassium, potentiation was similar to that observed in the absence of the former. In total, these findings provide novel information on potassium-mediated twitch potentiation.NEW & NOTEWORTHY Here, we investigated the length-dependence of twitch force potentiation by extracellular potassium in mouse extensor digitorum longus (EDL) in vitro, at 25°C. Potentiation magnitude did not display a statistically significant difference between the examined muscle lengths. These results describe, for the first time, the relationship of this form of potentiation with muscle length, thus furthering the understanding of how it is integrated in in vivo muscle function.

Effects of postactivation potentiation on mechanical output and muscle architecture during electrically induced contractions in plantar flexors.

This study investigated the effects of postactivation potentiation (PAP) on the force output and muscle architecture in plantar flexor muscles. The mechanical response to a single electrical stimulus (twitch) and to two (doublet) and three (triplet) stimuli (10-ms interpulse interval) was measured before and after a 6-s maximal voluntary contraction (MVC). Ultrasound imaging was used to measure fascicle length and pennation angle of the gastrocnemius medialis at rest and during the electrically induced contractions. Immediately after the conditioning MVC, twitch peak force (+40%) and its maximal rate of force development (+57%) and relaxation (+62%) were greater than before the MVC (P < 0.001). The PAP extent was less for the doublet than for the twitch and for the triplet than for the doublet (P < 0.05). Whereas none of the architectural parameters changed at rest, fascicle shortening and increase in pennation angle during contractions were greater after than before the conditioning MVC, with a greater extent (P < 0.001) during the twitch (+28% and +58%, respectively) compared with the doublet (+16% and +36%) and the triplet (+12% and +14%). Overall, our results indicate that the effect of the conditioning MVC on mechanical output and muscle architecture decreased from the twitch to the triplet in plantar flexor muscles. The decreased PAP observed during doublet and triplet compared with twitch indicates that the benefit of this mechanism to the enhancement of muscle performance becomes progressively less effective during successive muscle activation.NEW & NOTEWORTHY Postactivation potentiation (PAP), a form of activity-dependent potentiation, is defined as the acute enhancement of muscle contractile properties induced by a conditioning contraction, usually a maximal voluntary contraction. The purpose of our study was to investigate the effects of PAP on force production and muscle architecture in plantar flexor (PF) muscles. The results indicate that the extent of PAP decreased from the twitch to the triplet in PF muscles. Furthermore, this study shows that fascicle shortening and the increase in pennation angle were greater after the maximal voluntary contraction for the twitch, the doublet, and the triplet, with more pronounced effects for the twitch.

Force\u2013frequency relationship during fatiguing contractions of rat medial gastrocnemius muscle

The force–frequency relationship presents the amount of force a muscle can produce as a function of the frequency of activation. During repetitive muscular contractions, fatigue and potentiation may both impact the resultant contractile response. However, both the apparent fatigue observed, and the potential for activity-dependent potentiation can be affected by the frequency of activation. Thus, we wanted to explore the effects that repetitive stimulation had on the force–frequency relationship. The force–frequency relationship of the rat medial gastrocnemius muscle was investigated during consecutive bouts of increasing fatigue with 20 to 100 Hz stimulation. Force was measured prior to the fatiguing protocol, during each of three levels of fatigue, and after 30 min of recovery. Force at each frequency was quantified relative to the pre-fatigued 100 Hz contractions, as well as the percentage reduction of force from the pre-fatigued level at a given frequency. We observed less reduction in force at low frequencies compared to high frequencies, suggesting an interplay of fatigue and potentiation, in which potentiation can “protect” against fatigue in a frequency-dependent manner. The exact mechanism of fatigue is unknown, however the substantial reduction of force at high frequency suggests a role for reduced force per cross-bridge.

Calcium-induced calcium release and type 3 ryanodine receptors modulate the slow afterhyperpolarising current, sIAHP, and its potentiation in hippocampal pyramidal neurons.

The slow afterhyperpolarising current, sIAHP, is a Ca2+-dependent current that plays an important role in the late phase of spike frequency adaptation. sIAHP is activated by voltage-gated Ca2+ channels, while the contribution of calcium from ryanodine-sensitive intracellular stores, released by calcium-induced calcium release (CICR), is controversial in hippocampal pyramidal neurons. Three types of ryanodine receptors (RyR1-3) are expressed in the hippocampus, with RyR3 showing a predominant expression in CA1 neurons. We investigated the specific role of CICR, and particularly of its RyR3-mediated component, in the regulation of the sIAHP amplitude and time course, and the activity-dependent potentiation of the sIAHP in rat and mouse CA1 pyramidal neurons. Here we report that enhancement of CICR by caffeine led to an increase in sIAHP amplitude, while inhibition of CICR by ryanodine caused a small, but significant reduction of sIAHP. Inhibition of ryanodine-sensitive Ca2+ stores by ryanodine or depletion by the SERCA pump inhibitor cyclopiazonic acid caused a substantial attenuation in the sIAHP activity-dependent potentiation in both rat and mouse CA1 pyramidal neurons. Neurons from mice lacking RyR3 receptors exhibited a sIAHP with features undistinguishable from wild-type neurons, which was similarly reduced by ryanodine. However, the lack of RyR3 receptors led to a faster and reduced activity-dependent potentiation of sIAHP. We conclude that ryanodine receptor-mediated CICR contributes both to the amplitude of the sIAHP at steady state and its activity-dependent potentiation in rat and mouse hippocampal pyramidal neurons. In particular, we show that RyR3 receptors play an essential and specific role in shaping the activity-dependent potentiation of the sIAHP. The modulation of activity-dependent potentiation of sIAHP by RyR3-mediated CICR contributes to plasticity of intrinsic neuronal excitability and is likely to play a critical role in higher cognitive functions, such as learning and memory.

Synergistic excitability plasticity in cerebellar functioning

The cerebellum, a universal processor for sensory acquisition and internal models, and its association with synaptic and nonsynaptic plasticity have been envisioned as the biological correlates of learning, perception, and even thought. Indeed, the cerebellum is no longer considered merely as the locus of motor coordination and its learning. Here, we introduce the mechanisms underlying the induction of multiple types of plasticity in cerebellar circuit and give an overview focusing on the plasticity of nonsynaptic intrinsic excitability. The discovery of long-term potentiation of synaptic responsiveness in hippocampal neurons led investigations into changes of their intrinsic excitability. This activity-dependent potentiation of neuronal excitability is distinct from that of synaptic efficacy. Systematic examination of excitability plasticity has indicated that the modulation of various types of Ca2+ - and voltage-dependent K+ channels underlies the phenomenon, which is also triggered by immune activity. Intrinsic plasticity is expressed specifically on dendrites and modifies the integrative processing and filtering effect. In Purkinje cells, modulation of the discordance of synaptic current on soma and dendrite suggested a novel type of cellular learning mechanism. This property enables a plausible synergy between synaptic efficacy and intrinsic excitability, by amplifying electrical conductivity and influencing the polarity of bidirectional synaptic plasticity. Furthermore, the induction of intrinsic plasticity in the cerebellum correlates with motor performance and cognitive processes, through functional connections from the cerebellar nuclei to neocortex and associated regions: for example, thalamus and midbrain. Taken together, recent advances in neuroscience have begun to shed light on the complex functioning of nonsynaptic excitability and the synergy.

Localized Calcium Signaling and the Control of Coupling at Cx36 Gap Junctions.

A variety of electrical synapses are capable of activity-dependent plasticity, including both activity-dependent potentiation and activity-dependent depression. In several types of neurons, activity-dependent electrical synapse plasticity depends on changes in the local Ca2+ environment. To enable study of local Ca2+ signaling that regulates plasticity, we developed a GCaMP Ca2+ biosensor fused to the electrical synapse protein Connexin 36 (Cx36). Cx36-GCaMP transfected into mammalian cell cultures formed gap junctions at cell-cell boundaries and supported Neurobiotin tracer coupling that was regulated by protein kinase A signaling in the same way as Cx36. Cx36-GCaMP gap junctions robustly reported local Ca2+ increases in response to addition of a Ca2+ ionophore with increases in fluorescence that recovered during washout. Recovery was strongly dependent on Na+-Ca2+ exchange activity. In cells transfected with NMDA receptor subunits, Cx36-GCaMP revealed transient and concentration-dependent increases in local Ca2+ on brief application of glutamate. In HeLa cells, glutamate application increased Cx36-GCaMP tracer coupling through a mechanism that depended in part on Ca2+, calmodulin-dependent protein kinase II (CaMKII) activity. This potentiation of coupling did not require exogenous expression of glutamate receptors, but could be accomplished by endogenously expressed glutamate receptors with pharmacological characteristics reminiscent of NMDA and kainate receptors. Analysis of RNA Sequencing data from HeLa cells confirmed expression of NMDA receptor subunits NR1, NR2C, and NR3B. In summary, Cx36-GCaMP is an effective tool to measure changes in the Ca2+ microenvironment around Cx36 gap junctions. Furthermore, HeLa cells can serve as a model system to study glutamate receptor-driven potentiation of electrical synapses.

Structure and plasticity of silent synapses in developing hippocampal neurons visualized by super-resolution imaging

Excitatory synapses in the mammalian brain exhibit diverse functional properties in transmission and plasticity. Directly visualizing the structural correlates of such functional heterogeneity is often hindered by the diffraction-limited resolution of conventional optical imaging techniques. Here, we used super-resolution stochastic optical reconstruction microscopy (STORM) to resolve structurally distinct excitatory synapses formed on dendritic shafts and spines. The majority of these shaft synapses contained N-methyl-d-aspartate receptors (NMDARs) but not α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs), suggesting that they were functionally silent. During development, as more spine synapses formed with increasing sizes and expression of AMPARs and NMDARs, shaft synapses exhibited moderate reduction in density with largely unchanged sizes and receptor expression. Furthermore, upon glycine stimulation to induce chemical long-term potentiation (cLTP), the previously silent shaft synapses became functional shaft synapses by recruiting more AMPARs than did spine synapses. Thus, silent shaft synapse may represent a synaptic state in developing neurons with enhanced capacity of activity-dependent potentiation.

Activity-Dependent Plasticity at Associative Memory Cells in the Prefrontal Cortex

In addition to structural and functional alternations of neurons during their recruitment to be associative memory cells, these associative memory cells undergo the functional plasticity in response to their activity, called as recruitment-relevant plasticity and activity-dependent plasticity, respectively (Associative Memory Cells: Basic Units of Memory Trace by Jin-Hui Wang 2019). Here, we report the features of activity-dependent plasticity at associative memory cells studied by two-photon cell imaging. After the learning of triple associated signals (pair-stimulations by odor, whisker tactile and tail warming-up), secondary associative memory cells in response to these signals are recruited in the prefrontal cortex. The portion of functional associative memory cells is upregulated by intensive activity in one of three synaptic inputs or their combinations. When whisker, odor or tail stimulus is given intensively, the activity levels of these associative memory cells show low-to-high or high-to-low in response to original cues, especially activity-dependent depression in response to homosynaptic signal and activity-dependent potentiation in response to heterosynaptic signal. When whisker, odor and tail stimulations are given simultaneously, these associative memory cells express activity-dependent potentiation in response to all these input signals. The upregulated activities of norepinephrine and serotonin in the central nervous system strengthen the recruitment, the functional strength and the activity-dependent plasticity of associative memory cell in response to homosynaptic and heterosynaptic signals. As activity-dependent plasticity at associative memory cells occurs immediately after their activation, our results may be due to a possibility that the new synapses formed on associative memory cells during associative learning can be upregulated and downregulated between active state and inactive state, in which their intensive activities and monoamine neurotransmitters are critical modulators. The results can explain activity-related memory strengthening and the role of the arousal system through norepinephrine and serotonin in memory.

Calcium sensitivity during staircase with sequential incompletely fused contractions.

Activity dependent potentiation is thought to result from phosphorylation of the regulatory light chains of myosin, increasing Ca2+ sensitivity. Yet, Ca2+ sensitivity decreases early in a period of intermittent contractions. The purpose of this study was to investigate the early change in Ca2+ sensitivity during intermittent submaximal tetanic contractions. Flexor digitorum brevis muscle fibres were dissected from mice after cervical disarticulation. Fibres were superfused with Tyrode solution at 32 °C. Length was set to yield maximal tetanic force. Indo-1 was microinjected into fibres and allowed to dissipate for 30 min. Fluorescence was measured at 405 and 495 nm wavelength and the ratio was used to estimate [Ca2+]. A control force-Ca2+ relationship was determined with stimulation over a range of frequencies, yielding constants for slope, max force, and half-maximal [Ca2+] (pCa2 +50). Data were collected for sequential contractions at 40 Hz at 2 s intervals. Active force decreased over the first 1-4 contractions then increased. A force-pCa2+ curve was fit to each contraction, using the control values for the Hill slope and max force by adjusting pCa2+50 until the curve passed through the target contraction. Data are presented for three contractions for each fibre: first, maximum shift to the right, and last contraction. There was a significant shift to the right for pCa2+50 (decreased Ca2+ sensitivity), usually early in the series of intermittent contractions, then pCa2 +50 shifted to the left, but remained significantly different from the control value. Although potentiation is associated with increased Ca2+ sensitivity, this increase begins only after Ca2+ sensitivity has decreased and, in most cases, Ca2+ sensitivity does not increase above the control level.

Latent Sex Differences in Molecular Signaling That Underlies Excitatory Synaptic Potentiation in the Hippocampus.

Excitatory synapses can be potentiated by chemical neuromodulators, including 17β-estradiol (E2), or patterns of synaptic activation, as in long-term potentiation (LTP). Here, we investigated kinases and calcium sources required for acute E2-induced synaptic potentiation in the hippocampus of each sex and tested whether sex differences in kinase signaling extend to LTP. We recorded EPSCs from CA1 pyramidal cells in hippocampal slices from adult rats and used specific inhibitors of kinases and calcium sources. This revealed that, although E2 potentiates synapses to the same degree in each sex, cAMP-activated protein kinase (PKA) is required to initiate potentiation only in females. In contrast, mitogen-activated protein kinase, Src tyrosine kinase, and rho-associated kinase are required for initiation in both sexes; similarly, Ca2+/calmodulin-activated kinase II is required for expression/maintenance of E2-induced potentiation in both sexes. Calcium source experiments showed that L-type calcium channels and calcium release from internal stores are both required for E2-induced potentiation in females, whereas in males, either L-type calcium channel activation or calcium release from stores is sufficient to permit potentiation. To investigate the generalizability of a sex difference in the requirement for PKA in synaptic potentiation, we tested how PKA inhibition affects LTP. This showed that, although the magnitude of both high-frequency stimulation-induced and pairing-induced LTP is the same between sexes, PKA is required for LTP in females but not males. These results demonstrate latent sex differences in mechanisms of synaptic potentiation in which distinct molecular signaling converges to common functional endpoints in males and females.SIGNIFICANCE STATEMENT Chemical- and activity-dependent neuromodulation alters synaptic strength in both male and female brains, yet few studies have compared mechanisms of neuromodulation between the sexes. Here, we studied molecular signaling that underlies estrogen-induced and activity-dependent potentiation of excitatory synapses in the hippocampus. We found that, despite similar magnitude increases in synaptic strength in males and females, the roles of cAMP-regulated protein kinase, internal calcium stores, and L-type calcium channels differ between the sexes. Therefore, latent sex differences in which the same outcome is achieved through distinct underlying mechanisms in males and females include kinase and calcium signaling involved in synaptic potentiation, demonstrating that sex is an important factor in identification of molecular targets for therapeutic development based on mechanisms of neuromodulation.

Dopaminergic innervation and modulation of hippocampal networks.

The catecholamine dopamine plays an important role in hippocampus-dependent plasticity and related learning and memory processes. Dopamine secretion in the hippocampus is activated by, e.g., salient or novel stimuli, thereby helping to establish and to stabilize hippocampus-dependent memories. Disturbed dopaminergic function in the hippocampus leads to severe pathophysiological conditions. While the role and importance of dopaminergic modulation of hippocampal networks have been unequivocally proven, there is still a lack of detailed molecular and cellular mechanistic understanding of how dopamine orchestrates these hippocampal processes. In this chapter of the special issue "Hippocampal structure and function," we will discuss the current understanding of dopaminergic modulation of basal synaptic transmission and long-lasting, activity-dependent potentiation or depression.

Use-dependent potentiation of voltage-gated calcium channels rescues neurotransmission in nerve terminals intoxicated by botulinum neurotoxin serotype A

Botulinum neurotoxins (BoNTs) are highly potent toxins that cleave neuronal SNARE proteins required for neurotransmission, causing flaccid paralysis and death by asphyxiation. Currently, there are no clinical treatments to delay or reverse BoNT-induced blockade of neuromuscular transmission. While aminopyridines have demonstrated varying efficacy in transiently reducing paralysis following BoNT poisoning, the precise mechanisms by which aminopyridines symptomatically treat botulism are not understood. Here we found that activity-dependent potentiation of presynaptic voltage-gated calcium channels (VGCCs) underlies 3,4-diaminopyridine (3,4-DAP)-mediated rescue of neurotransmission in central nervous system synapses and mouse diaphragm neuromuscular junctions fully intoxicated by BoNT serotype A. Combinatorial treatments with 3,4-DAP and VGCC agonists proved synergistic in restoring suprathreshold endplate potentials in mouse diaphragms fully intoxicated by BoNT/A. In contrast, synapses fully intoxicated by BoNT serotypes D or E were refractory to synaptic rescue by any treatment. We interpret these data to propose that increasing the duration or extent of VGCC activation prolongs the opportunity for low-efficiency fusion by fusogenic complexes incorporating BoNT/A-cleaved SNAP-25. The identification of VGCC agonists that rescue neurotransmission in BoNT/A-intoxicated synapses provides compelling evidence for potential therapeutic utility in some cases of human botulism.

A Cooperative Mechanism Involving Ca2+-Permeable AMPA Receptors and Retrograde Activation of GABAB Receptors in Interpeduncular Nucleus Plasticity

The medial habenula-interpeduncular nucleus (MHb-IPN) pathway, which connects the limbic forebrain to the midbrain, has recently been implicated in aversive behaviors. The MHb-IPN circuit is characterized by a unique topographical organization, an excitatory role of GABA, and a prominent co-release of neurotransmitters and neuropeptides. However, little is known about synaptic plasticity in this pathway. An application of a high-frequency stimulation resulted in a long-lasting potentiation of glutamate release in IPN neurons. Our experiments reveal that a Ca2+-permeable AMPA receptor (CPAR)-dependent release of GABA from IPN neurons and a retrograde activation of GABAB receptors on MHb terminals result in a long-lasting enhancement of glutamate release. Strikingly, adolescent IPN neurons lacked CPARs and exhibited an inability to undergo plasticity. In addition, fear conditioning suppressed an activity-dependent potentiation of MHb-IPN synapses, whereas fear extinction reversed this plasticity deficit, suggesting a role of the MHb-IPN synaptic plasticity in the regulation of aversive behaviors.

TRPV1 Nociceptor Activity Initiates USP5/T-type Channel-Mediated Plasticity

Peripheral nerve injury and tissue inflammation resultin upregulation of the deubiquitinase USP5, thus causing a dysregulation of T-type calcium channel activity and increased pain sensitivity. Here, we have explored the role of afferent fiber activity in this process. Conditioning stimulation of optogenetically targeted cutaneous TRPV1 expressing nociceptors, but not that of non-nociceptive fibers, resulted in enhanced expression of USP5 in mouse dorsal root ganglia and spinal dorsal horn, along with decreased withdrawal thresholds for thermal and mechanical stimuli that abated after 24hr. This sensitization was drastically reduced by an interfering peptide that prevented USP5-Cav3.2 association. Sensitization was relieved by pharmacological block of TRPV1 afferents, but not of myelinated neurons. In spinal cord slice recordings, we could optogenetically trigger an activity-dependent potentiation of presynaptic neurotransmission in the spinal dorsal horn that relied on Cav3.2 channel activity. This neuronal-activity-induced USP5 upregulation may underlie a protective, transient sensitization of the pain pathway.

Impaired neuronal operation through aberrant intrinsic plasticity in epilepsy.

Patients with temporal lobe epilepsy often display cognitive comorbidity with recurrent seizures. However, the cellular mechanisms underlying the impairment of neuronal information processing remain poorly understood in temporal lobe epilepsy. Within the hippocampal formation neuronal networks undergo major reorganization, including the sprouting of mossy fibers in the dentate gyrus; they establish aberrant recurrent synapses between dentate granule cells and operate via postsynaptic kainate receptors. In this report, we tested the hypothesis that this aberrant local circuit alters information processing of perforant path inputs constituting the major excitatory afferent pathway from entorhinal cortex to dentate granule cells. Experiments were performed in dentate granule cells from control rats and rats with temporal lobe epilepsy induced by pilocarpine hydrochloride treatment. Neurons were recorded in patch clamp in whole cell configuration in hippocampal slices. Our present data revealed that an aberrant readout of synaptic inputs by kainate receptors triggered a long-lasting impairment of the perforant path input-output operation in epileptic dentate granule cells. We demonstrated that this is due to the aberrant activity-dependent potentiation of the persistent sodium current altering intrinsic firing properties of dentate granule cells. We propose that this aberrant activity-dependent intrinsic plasticity, which lastingly impairs the information processing of cortical inputs in dentate gyrus, may participate in hippocampal-related cognitive deficits, such as those reported in patients with epilepsy.

Low Dose ZD7288 Attenuates the Ischemia/Reperfusion-Induced Impairment of Long-Term Potentiation Induction at Hippocampal Schaffer Collateral-CA1 Synapses

Focal cerebral ischemia can impair the induction of activity-dependent long-term potentiation (LTP) in the hippocampus. This impairment of hippocampal synaptic plasticity can be caused by excitotoxicity and subsequent perturbation of hippocampal LTP-relevant transmitter systems, which include NR2B and PSD-95. It has been suggested that hyperpolarization-activated cyclic-nucleotide-gated (HCN) channels may play an important role in the control of membrane excitability and rhythmic neuronal activity. Our previous study has indicated that the selective HCN channel blocker ZD7288 can produce a dose-dependent inhibition of the induction of LTP at the Schaffer collateral-CA1 synapse of hippocampus by reducing the amount of glutamate released. It has also been demonstrated that ZD7288 can protect against neuronal injury caused by oxygen glucose deprivation. In the present study, we investigated the effect of ZD7288 on the induction of activity-dependent LTP and the expression of NR2B and PSD-95 after focal cerebral ischemia/reperfusion injury. The results showed that the induction of LTP was significantly impaired and the levels of NR2B and PSD-95 mRNA and protein were markedly decreased in the CA1 region of hippocampus following focal cerebral ischemia/reperfusion injury. Administration of low dose ZD7288 (0.25 μg) at 30 min and 3 h after the onset of ischemia attenuated the impairment of LTP induction and alleviated the NR2B and PSD-95 mRNA and protein down-regulation commonly induced by cerebral ischemia/reperfusion injury. These results suggest that low dose ZD7288 can ameliorate the ischemia/reperfusion-induced impairment of synaptic plasticity in the hippocampal CA1 region.

Activity-Dependent Potentiation of an Asynchronous Component of GABA-ergic Synaptic Currents in Cultured Hippocampal Neurons

After stimulation of GABA-ergic synaptic connections in a culture of hippocampal neurons by a series of depolarizing current pulses (5-45 sec-1, 5-20 stimuli), an inhibitory postsynaptic current (IPSC) consisting of a phasic component and an asynchronous component develops in the postsynaptic neuron. In experiments with simultaneous recording from two synapticaly connected neurons, we found that 2-min-long depolarization of the presynaptic cell to +40 mV also led to the development of the asynchronous IPSC component (аIPSC) induced by a single action potential (AP) in the presynaptic neuron. In this case, the decay time of IPSC was, on average, three times greater than in the first case. Analogous potentiation of аIPSC was observed in a part of the synaptic connections after their longlasting (15 min) stimulation by series of depolarizing current pulses (45 sec-1, 20 stimuli, betweenseries intervals 23 sec). Such potentiation of аIPSC was preserved for a long time (from 3 to 17 min). Simultaneously with potentiation of аIPSC, we observed potentiation of plateau-like inward current through the membrane of the presynaptic neuron in some synaptic connections. The charge transferred by this current through the membrane of the presynaptic cell correlated significantly with the decay time of evoked IPSC in the postsynaptic neuron (mean correlation coefficient 0.83±0.10). This fact can be considered an indication that the above-mentioned inward current mediates potentiation of the above IPSC. The obtained data allow us to hypothesize that the above-described form of plasticity can regulate the efficacy of GABA-ergic synaptic transmission in the hippocampus in the physiological modes of synaptic activity.

Tetanic force potentiation of mouse fast muscle is shortening speed dependent

The activity dependent potentiation of peak isometric force associated with phosphorylation of the myosin regulatory light chain (RLC) is generally restricted to low activation frequencies. The purpose of this study was to determine if muscle shortening speed influenced the stimulus frequency domain over which concentric force potentiation was observed. To this end, mouse extensor digitorum longus (EDL) muscles (in vitro, 25 °C) were activated at a range of test frequencies (10, 25, 45, 70 or 100 Hz) during shortening ramps at 0.10, 0.30 or 0.50 of the maximal velocity of shortening (V(max)). This procedure was performed before and after a standard conditioning stimulus (CS) that elevated RLC phosphorylation from 0.08 ± 0.01 (rest) to 0.55 ± 0.01 (stimulated) moles phosphate per mol RLC, respectively (n = 9-11) (P < 0.01). When data from all test frequencies were collapsed, the CS potentiated mean concentric force at 0.10, 0.30 and 0.50 V(max) to 1.02 ± 0.03, 1.37 ± 0.03 and 1.59 ± 0.05 of unpotentiated, pre-CS values, respectively (n = 8, P < 0.05). In addition, increasing shortening speed also increased the activation frequency at which concentric force potentiation was maximal, i.e. from 10 Hz at 0.10 V(max) to 10-25 and 25-45 Hz at 0.30 and 0.50 V(max), respectively. These results indicate that both the magnitude of and activation frequency dependence for concentric force potentiation of mouse EDL muscle is shortening speed dependent. Thus, muscle shortening speed may be a critical factor determining the functional utility of the myosin RLC phosphorylation mechanism.

Two independent forms of activity-dependent potentiation regulate electrical transmission at mixed synapses on the Mauthner cell

Mixed (electrical and chemical) synaptic contacts on the Mauthner cells, known as Club endings, constitute a valuable model for the study of vertebrate electrical transmission. While electrical synapses are still perceived by many as passive intercellular channels that lack modifiability, a wealth of experimental evidence shows that gap junctions at Club endings are subject to dynamic regulatory control by two independent activity-dependent mechanisms that lead to potentiation of electrical transmission. One of those mechanisms relies on activation of NMDA receptors and postsynaptic CaMKII. A second mechanism relies on mGluR activation and endocannabinoid production and is indirectly mediated via the release of dopamine from nearby varicosities, which in turn leads to potentiation of the synaptic response via a PKA-mediated postsynaptic mechanism. We review here these two forms of potentiation and their signaling mechanisms, which include the activation of two kinases with well-established roles as regulators of synaptic strength, as well as the functional implications of these two forms of potentiation. Special Issue entitled Electrical Synapses.

Restoration of Contralateral Representation in the Mouse Somatosensory Cortex after Crossing Nerve Transfer

Avulsion of spinal nerve roots in the brachial plexus (BP) can be repaired by crossing nerve transfer via a nerve graft to connect injured nerve ends to the BP contralateral to the lesioned side. Sensory recovery in these patients suggests that the contralateral primary somatosensory cortex (S1) is activated by afferent inputs that bypassed to the contralateral BP. To confirm this hypothesis, the present study visualized cortical activity after crossing nerve transfer in mice through the use of transcranial flavoprotein fluorescence imaging. In naïve mice, vibratory stimuli applied to the forepaw elicited localized fluorescence responses in the S1 contralateral to the stimulated side, with almost no activity in the ipsilateral S1. Four weeks after crossing nerve transfer, forepaw stimulation in the injured and repaired side resulted in cortical responses only in the S1 ipsilateral to the stimulated side. At eight weeks after crossing nerve transfer, forepaw stimulation resulted in S1 cortical responses of both hemispheres. These cortical responses were abolished by cutting the nerve graft used for repair. Exposure of the ipsilateral S1 to blue laser light suppressed cortical responses in the ipsilateral S1, as well as in the contralateral S1, suggesting that ipsilateral responses propagated to the contralateral S1 via cortico-cortical pathways. Direct high-frequency stimulation of the ipsilateral S1 in combination with forepaw stimulation acutely induced S1 bilateral cortical representation of the forepaw area in naïve mice. Cortical responses in the contralateral S1 after crossing nerve transfer were reduced in cortex-restricted heterotypic GluN1 (NMDAR1) knockout mice. Functional bilateral cortical representation was not clearly observed in genetically manipulated mice with impaired cortico-cortical pathways between S1 of both hemispheres. Taken together, these findings strongly suggest that activity-dependent potentiation of cortico-cortical pathways has a critical role for sensory recovery in patients after crossing nerve transfer.