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1. Novel types of frequency filtering in the lateral perforant path projections to dentate gyrus

   https://doi.org/10.1113/JP283012  

   Quintanilla, Julian ; Jia, Yousheng ; Lauterborn, Julie C. ; Pruess, Benedict S. ; Le, Aliza A. ; Cox, Conor D. ; Gall, Christine M. ; Lynch, Gary ; Gunn, Benjamin G. ( August 2022 , The Journal of Physiology)

   Despite its evident importance to learning theory and models, the manner in which the lateral perforant path (LPP) transforms signals from entorhinal cortex to hippocampus is not well understood. The present studies measured synaptic responses in the dentate gyrus (DG) of adult mouse hippocampal slices during different patterns of LPP stimulation. Theta (5 Hz) stimulation produced a modest within‐train facilitation that was markedly enhanced at the level of DG output. Gamma (50 Hz) activation resulted in a singular pattern with initial synaptic facilitation being followed by a progressively greater depression. DG output was absent after only two pulses. Reducing release probability with low extracellular calcium instated frequency facilitation to gamma stimulation while long‐term potentiation, which increases release by LPP terminals, enhanced within‐train depression. Relatedly, per terminal concentrations of VGLUT2, a vesicular glutamate transporter associated with high release probability, were much greater in the LPP than in CA3–CA1 connections. Attempts to circumvent the potent gamma filter using a series of short (three‐pulse) 50 Hz trains spaced by 200 ms were only partially successful: composite responses were substantially reduced after the first burst, an effect opposite to that recorded in field CA1. The interaction between bursts was surprisingly persistent (>1.0 s). Low calcium improved throughput during theta/gamma activation but buffering of postsynaptic calcium did not. In all, presynaptic specializations relating to release probability produce an unusual but potent type of frequency filtering in the LPP. Patterned burst input engages a different type of filter with substrates that are also likely to be located presynaptically.image

   The lateral perforant path (LPP)–dentate gyrus (DG) synapse operates as a low‐pass filter, where responses to a train of 50 Hz, γ frequency activation are greatly suppressed.

   Activation with brief bursts of γ frequency information engages a secondary filter that persists for prolonged periods (lasting seconds).

   Both forms of LPP frequency filtering are influenced by presynaptic, as opposed to postsynaptic, processes; this contrasts with other hippocampal synapses.

   LPP frequency filtering is modified by the unique presynaptic long‐term potentiation at this synapse.

   Computational simulations indicate that presynaptic factors associated with release probability and vesicle recycling may underlie the potent LPP–DG frequency filtering.

    

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2. Violation of the ultrastructural size principle in the dorsolateral prefrontal cortex underlies working memory impairment in the aged common marmoset (Callithrix jacchus)

   https://doi.org/10.3389/fnagi.2023.1146245  

   Glavis-Bloom, Courtney ; Vanderlip, Casey R. ; Weiser Novak, Sammy ; Kuwajima, Masaaki ; Kirk, Lyndsey ; Harris, Kristen M. ; Manor, Uri ; Reynolds, John H. ( April 2023 , Frontiers in Aging Neuroscience)

   Morphology and function of the dorsolateral prefrontal cortex (dlPFC), and corresponding working memory performance, are affected early in the aging process, but nearly half of aged individuals are spared of working memory deficits. Translationally relevant model systems are critical for determining the neurobiological drivers of this variability. The common marmoset (Callithrix jacchus) is advantageous as a model for these investigations because, as a non-human primate, marmosets have a clearly defined dlPFC that enables measurement of prefrontal-dependent cognitive functions, and their short (∼10 year) lifespan facilitates longitudinal studies of aging. Previously, we characterized working memory capacity in a cohort of marmosets that collectively covered the lifespan, and found age-related working memory impairment. We also found a remarkable degree of heterogeneity in performance, similar to that found in humans. Here, we tested the hypothesis that changes to synaptic ultrastructure that affect synaptic efficacy stratify marmosets that age with cognitive impairment from those that age without cognitive impairment. We utilized electron microscopy to visualize synapses in the marmoset dlPFC and measured the sizes of boutons, presynaptic mitochondria, and synapses. We found that coordinated scaling of the sizes of synapses and mitochondria with their associated boutons is essential for intact working memory performance in aged marmosets. Further, lack of synaptic scaling, due to a remarkable failure of synaptic mitochondria to scale with presynaptic boutons, selectively underlies age-related working memory impairment. We posit that this decoupling results in mismatched energy supply and demand, leading to impaired synaptic transmission. We also found that aged marmosets have fewer synapses in dlPFC than young, though the severity of synapse loss did not predict whether aging occurred with or without cognitive impairment. This work identifies a novel mechanism of synapse dysfunction that stratifies marmosets that age with cognitive impairment from those that age without cognitive impairment. The process by which synaptic scaling is regulated is yet unknown and warrants future investigation. 

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3. Postsynaptic Calcium Extrusion at the Mouse Neuromuscular Junction Alkalinizes the Synaptic Cleft

   https://doi.org/10.1523/JNEUROSCI.0815-23.2023  

   Durbin, Ryan J. ; Heredia, Dante J. ; Gould, Thomas W. ; Renden, Robert B. ( July 2023 , The Journal of Neuroscience)

   Neurotransmission is shaped by extracellular pH. Alkalization enhances pH-sensitive transmitter release and receptor activation, whereas acidification inhibits these processes and can activate acid-sensitive conductances in the synaptic cleft. Previous work has shown that the synaptic cleft can either acidify because of synaptic vesicular release and/or alkalize because of Ca²⁺extrusion by the plasma membrane ATPase (PMCA). The direction of change differs across synapse types. At the mammalian neuromuscular junction (NMJ), the direction and magnitude of pH transients in the synaptic cleft during transmission remain ambiguous. We set out to elucidate the extracellular pH transients that occur at this cholinergic synapse under near-physiological conditions and identify their sources. We monitored pH-dependent changes in the synaptic cleft of the mouse levator auris longus using viral expression of the pseudoratiometric probe pHusion-Ex in the muscle. Using mice from both sexes, a significant and prolonged alkalization occurred when stimulating the connected nerve for 5 s at 50 Hz, which was dependent on postsynaptic intracellular Ca²⁺release. Sustained stimulation for a longer duration (20 s at 50 Hz) caused additional prolonged net acidification at the cleft. To investigate the mechanism underlying cleft alkalization, we used muscle-expressed GCaMP3 to monitor the contribution of postsynaptic Ca²⁺. Activity-induced liberation of intracellular Ca²⁺in muscle positively correlated with alkalization of the synaptic cleft, whereas inhibiting PMCA significantly decreased the extent of cleft alkalization. Thus, cholinergic synapses of the mouse NMJ typically alkalize because of cytosolic Ca²⁺liberated in muscle during activity, unless under highly strenuous conditions where acidification predominates.

   SIGNIFICANCE STATEMENTChanges in synaptic cleft pH alter neurotransmission, acting on receptors and channels on both sides of the synapse. Synaptic acidification has been associated with a myriad of diseases in the central and peripheral nervous system. Here, we report that in near-physiological recording conditions the cholinergic neuromuscular junction shows use-dependent bidirectional changes in synaptic cleft pH—immediate alkalinization and a long-lasting acidification under prolonged stimulation. These results provide further insight into physiologically relevant changes at cholinergic synapses that have not been defined previously. Understanding and identifying synaptic pH transients during and after neuronal activity provides insight into short-term synaptic plasticity synapses and may identify therapeutic targets for diseases.

    

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4. ATP diffusional gradients are sufficient to maintain bioenergetic homeostasis in synaptic boutons lacking mitochondria

   https://doi.org/10.1002/cnm.3696  

   Kuznetsov, Ivan A. ; Kuznetsov, Andrey V. ( March 2023 , International Journal for Numerical Methods in Biomedical Engineering)

   Previous work on mitochondrial distribution in axons has shown that approximately half of the presynaptic release sites do not contain mitochondria, raising the question of how the boutons that do not contain mitochondria are supplied with ATP. Here, we develop and apply a mathematical model to study this question. Specifically, we investigate whether diffusive transport of ATP is sufficient to support the exocytic functionality in synaptic boutons which lack mitochondria. Our results demonstrate that the difference in ATP concentration between a bouton containing a mitochondrion and a neighboring bouton lacking a mitochondrion is only approximately 0.4%, which is still 3.75 times larger than the ATP concentration minimally required to support synaptic vesicle release. This work therefore suggests that passive diffusion of ATP is sufficient to maintain the functionality of boutons which do not contain mitochondria.

    

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5. Mitochondrial phosphagen kinases support the volatile power demands of motor nerve terminals

   https://doi.org/10.1113/JP284872  

   Justs, Karlis A. ; Sempertegui, Sergio ; Riboul, Danielle V. ; Oliva, Carlos D. ; Durbin, Ryan J. ; Crill, Sarah ; Stawarski, Michal ; Su, Chenchen ; Renden, Robert B. ; Fily, Yaouen ; et al ( November 2023 , The Journal of Physiology)

   Motor neurons are the longest neurons in the body, with axon terminals separated from the soma by as much as a meter. These terminals are largely autonomous with regard to their bioenergetic metabolism and must burn energy at a high rate to sustain muscle contraction. Here, through computer simulation and drawing on previously published empirical data, we determined that motor neuron terminals inDrosophila larvae experience highly volatile power demands. It might not be surprising then, that we discovered the mitochondria in the motor neuron terminals of bothDrosophila and mice to be heavily decorated with phosphagen kinases ‐ a key element in an energy storage and buffering system well‐characterized in fast‐twitch muscle fibres. Knockdown of arginine kinase 1 (ArgK1) inDrosophilalarval motor neurons led to several bioenergetic deficits, including mitochondrial matrix acidification and a faster decline in the cytosol ATP to ADP ratio during axon burst firing.image

   Neurons commonly fire in bursts imposing highly volatile demands on the bioenergetic machinery that generates ATP.

   Using a computational approach, we built profiles of presynaptic power demand at the level of single action potentials, as well as the transition from rest to sustained activity.

   Phosphagen systems are known to buffer ATP levels in muscles and we demonstrate that phosphagen kinases, which support such phosphagen systems, also localize to mitochondria in motor nerve terminals of fruit flies and mice.

   By knocking down phosphagen kinases in fruit fly motor nerve terminals, and using fluorescent reporters of the ATP:ADP ratio, lactate, pH and Ca²⁺, we demonstrate a role for phosphagen kinases in stabilizing presynaptic ATP levels.

   These data indicate that the maintenance of phosphagen systems in motor neurons, and not just muscle, could be a beneficial initiative in sustaining musculoskeletal health and performance.

    

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