Biology subjects

Young, S. M.

Publications and source records attributed to Young, S. M..

8 recordsLinked to original sources

bioRxiv2026

Presynaptic CaV2.1 calcium dependent facilitation is essential for faithful auditory information transfer

Activity-dependent modulation of presynaptic voltage-gated Ca2+ channels (CaV2) regulates Ca2+ influx to control neuronal circuit output. Although CaV2.1 can undergo robust Ca2+ dependent facilitation (CDF), whether it occurs in native central nervous system neuronal circuits is disputed. Accurate auditory information processing requires precise and reliable synaptic transmission at high activity rates in the auditory brainstem. To determine if CaV2.1 CDF is a key regulator of high-fidelity synaptic transmission, we expressed CaV2.1 splice variants capable (CaV2.1 37a) or incapable (CaV2.1 37b) of CDF at the calyx of Held presynaptic terminal in the auditory brainstem. We found no difference in basal CaV2.1 currents or synaptic transmission. However, CaV2.1 37b terminals lacked CDF, synaptic facilitation and had a decreased reliability and precision of postsynaptic action-potential firing. Additionally, the Wave III amplitude of the auditory brainstem responses was reduced. We propose that CaV2.1 CDF is essential for accurate auditory information processing.

neuroscience↗

bioRxiv2025

Pre-Sensory Spontaneous Activity Accelerates Coordinated Maturation of Synaptic Partners and Drives Transition to the Mature Physiological Phenotype

Pre-sensory (ps), peripherally-generated spontaneous activity (SA) is important for establishing basic topography of central components of sensory pathways. However, roles for psSA in sculpting connections at the single neuron level and in driving functional maturation of postsynaptic targets are difficult to isolate and therefore little explored. We capitalized on the temporal onset of cochlea-generated psSA just prior to growth of the calyx of Held (CH), the largest nerve terminal in the mammalian brain, and its well-defined circuit topography to explore the causal roles for psSA in targeted synaptogenesis and postsynaptic functional maturation. To this end, we developed a viral vector to rapidly express tetanus neurotoxin (TeNT), which blocks neurotransmission, and selectively introduced it into CH-forming neurons at the onset of psSA. TeNT expression did not prevent but delayed and altered the developmental trajectory of neural circuit maturation, which in sum prevented the postsynaptic neuron from acquiring its mature phasic firing phenotype. Thus, psSA initiates and drives circuit maturation along central sensory pathways, but alternative mechanisms can partially compensate in its absence.

neuroscience↗

bioRxiv2025

Prevenying vision loss in a mouse model of Leber Congenital Amaurosis by engineered tRNA

Premature termination codons (PTCs) are associated with rare genetic disorders. Inducing targeted read-through of these nonsense mutations presents a potential therapeutic strategy for modifying disease outcomes. We previously reported that one such PTC, W53X, in the KCNJ13 gene causes blindness and Leber congenital amaurosis type-16 (LCA-16) due to loss of function of the inwardly rectifying potassium channel 7.1 (Kir7.1). Here, we present the proof of concept of a therapeutic approach based on anticodon-engineered transfer RNA (ACE-tRNA). The ACE-tRNA encodes the amino acid tryptophan (Trp) and suppresses the W53X PTC, restoring full-length protein expression. We used helper-dependent adenovirus (HDAd) to deliver the ACE-tRNATrp.UAG (tRNATrp.UAG) and rescue Kir7.1 function and physiology in patient-specific human induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE) cells. Furthermore, in a W53X mouse model of LCA16, HDAd delivery of tRNATrp.UAG resulted in durable restoration of vision as measured by retinography. This study provides the first example of the therapeutic application of ACE-tRNA for treating an inherited form of blindness.

physiology↗

bioRxiv2023

Fecal Dysbiosis and Inflammation in Intestinal-Specific Cftr Knockout Mice on Regimens Preventing Intestinal Obstruction

Chronic intestinal inflammation is a poorly understood manifestation of Cystic Fibrosis (CF), which may be refractory to ion channel CFTR modulator therapy. People with CF exhibit intestinal dysbiosis which has potential for stimulating intestinal and systemic inflammation. CFTR is expressed in organ epithelia and in the leukocyte population. Here, we investigate the contribution of intestinal epithelial-specific loss of Cftr (iCftr KO) to dysbiosis and inflammation in mice treated with either of two anti-obstructive dietary regimens necessary to maintain CF mouse models (PEG laxative or a liquid diet, LiqD). Feces collected from iCftr KO mice and their wildtype (WT) sex-matched littermates were used to measure fecal calprotectin and to perform 16S rRNA sequencing to characterize the gut microbiome. Fecal calprotectin was elevated in iCftr KO relative to WT samples of mice consuming either PEG or LiqD. PEG iCftr KO mice did not show a change in -diversity versus WT but demonstrated a significant difference in microbial composition ({beta}-diversity) with increases in phylum Proteobacteria, family Peptostreptococcaceae, four genera of Clostridia including C. innocuum, and mucolytic genus Akkermansia. Fecal microbiome analysis of LiqD iCftr KO mice showed both decreased -diversity and differences in microbial composition with increases in Proteobacteria family Enterobacteriaceae, Firmicutes families Clostridiaceae and Peptostreptococcaceae, and enrichment of Clostridium perfringens, C. innocuum, C. difficile, mucolytic Ruminococcus gnavus, and reduction of Akkermansia. It was concluded that epithelial-specific loss of Cftr is a major driver of CF intestinal dysbiosis and inflammation with significant similarities to previous studies of global Cftr KO mice. New and noteworthyChronic intestinal inflammation is a manifestation of cystic fibrosis (CF), a disease caused by loss of the anion channel CFTR that is expressed in many tissues. This study shows that intestinal epithelial cell-specific loss of CFTR (iCftr KO) in mice is sufficient to induce intestinal dysbiosis and inflammation. Studies were performed on mice consuming either dietary regimen (PEG laxative or liquid diet) routinely used to prevent obstruction in CF mice.

systems biology↗

bioRxiv2023

CaV2.1 α1 subunit motifs that control presynaptic CaV2.1 subtype abundance are distinct from CaV2.1 preference

Presynaptic voltage-gated Ca2+ channels (CaV) subtype abundance at mammalian synapses regulates synaptic transmission in health and disease. In the mammalian central nervous system, most presynaptic terminals are CaV2.1 dominant with a developmental reduction in CaV2.2 and CaV2.3 levels, and CaV2 subtype levels are altered in various diseases. However, the molecular mechanisms controlling presynaptic CaV2 subtype levels are largely unsolved. Since the CaV2 1 subunit cytoplasmic regions contain varying levels of sequence conservation, these regions are proposed to control presynaptic CaV2 subtype preference and abundance. To investigate the potential role of these regions, we expressed chimeric CaV2.1 1 subunits containing swapped motifs with the CaV2.2 and CaV2.3 1 subunit on a CaV2.1/CaV2.2 null background at the calyx of Held presynaptic terminal. We found that expression of CaV2.1 1 subunit chimeras containing the CaV2.3 loop II-III region or cytoplasmic C-terminus (CT) resulted in a large reduction of presynaptic Ca2+ currents compared to the CaV2.1 1 subunit. However, the Ca2+ current sensitivity to the CaV2.1 blocker Agatoxin-IVA, was the same between the chimeras and the CaV2.1 1 subunit. Additionally, we found no reduction in presynaptic Ca2+ currents with CaV2.1/2.2 cytoplasmic CT chimeras. We conclude that the motifs in the CaV2.1 loop II-III and CT do not individually regulate CaV2.1 preference, but these motifs control CaV2.1 levels and the CaV2.3 CT contains motifs that negatively regulate presynaptic CaV2.3 levels. We propose that the motifs controlling presynaptic CaV2.1 preference are distinct from those regulating CaV2.1 levels and may act synergistically to impact pathways regulating CaV2.1 preference and abundance. Key points summaryO_LIPresynaptic CaV2 subtype abundance regulates neuronal circuit properties, however the mechanisms regulating presynaptic CaV2 subtype abundance and preference remains enigmatic. C_LIO_LIThe CaV 1 subunit determines subtype and contains multiple motifs implicated in regulating presynaptic subtype abundance and preference. C_LIO_LIThe CaV2.1 1 subunit domain II-III loop and cytoplasmic C-terminus are positive regulators of presynaptic CaV2.1 abundance but do not regulate preference. C_LIO_LIThe CaV2.3 1 subunit cytoplasmic C-terminus negatively regulates presynaptic CaV2 subtype abundance but not preference while the CaV2.2 1 subunit cytoplasmic C-terminus is not a key regulator of presynaptic CaV2 subtype abundance or preference. C_LIO_LIThe CaV2 1 subunit motifs determining the presynaptic CaV2 preference are distinct from abundance. C_LI

neuroscience↗

bioRxiv2022

Presynaptic Rac1 controls synaptic strength through the regulation of synaptic vesicle priming.

Synapses contain a limited number of synaptic vesicles (SVs) that are released in response to action potentials (APs). Therefore, sustaining synaptic transmission over a wide range of AP firing rates and timescales depends on SV release and replenishment. Although actin dynamics impact synaptic transmission, how presynaptic regulators of actin signaling cascades control SV release and replenishment remains unresolved. Rac1, a Rho GTPase, regulates actin signaling cascades that control synaptogenesis, neuronal development, and postsynaptic function. However, the presynaptic role of Rac1 in regulating synaptic transmission is unclear. To unravel Rac1s roles in controlling transmitter release, we performed selective presynaptic ablation of Rac1 at the mature mouse calyx of Held synapse. Loss of Rac1 increased synaptic strength, accelerated EPSC recovery after conditioning stimulus trains, and augmented spontaneous SV release with no change in presynaptic morphology or AZ ultrastructure. Analyses with constrained short-term plasticity models revealed faster SV priming kinetics and, depending on model assumptions, elevated SV release probability or higher abundance of tightly docked fusion-competent SVs in Rac1-deficient synapses. We conclude that presynaptic Rac1 is a key regulator of synaptic transmission and plasticity mainly by regulating the dynamics of SV priming and potentially SV release probability.

neuroscience↗

bioRxiv2019

Presynaptic development is controlled by the core active zone proteins CAST/ELKS

Many presynaptic active zone proteins have multiple regulatory roles that vary during distinct stages of neuronal circuit development. However, our understanding how presynaptic active zone proteins regulate synapse development during neuronal circuit maturation is in its early stages. Although CAST/ELKS are presynaptic active zone core proteins, their roles in synapse development in the mammalian central nervous system remain enigmatic. To unravel CAST/ELKS roles in glutamatergic synapse development, we analyzed how their loss during the early stages of circuit maturation impacted the calyx of Held presynaptic terminal development and function. We found a reduction in presynaptic surface area and volume, but an increase in active zone size. Additionally, we found a reduction in all presynaptic Cav2 channel subtype currents. Surprisingly, these changes did not impair synaptic transmission. We propose that CAST/ELKS are involved in pathways regulating presynaptic morphological properties and Cav2 channel subtype levels during early stages of neuronal circuit maturation.

neuroscience↗

bioRxiv2019

Presynaptic mitochondria volumes and complexity of subsynaptic distribution increase during development at a high-fidelity synapse

The calyx of Held, a large glutamatergic presynaptic terminal in the auditory brainstem undergoes developmental changes to support the high action-potential firing rates required for auditory information encoding. In addition, calyx terminals are morphologically diverse which impacts vesicle release properties and synaptic plasticity. Mitochondria influence synaptic plasticity through calcium buffering and are crucial for providing the energy required for synaptic transmission. Therefore, it has been postulated that mitochondrial levels increase during development and contribute to the morphological-functional diversity in the mature calyx. However, the developmental profile of mitochondrial volumes and subsynaptic distribution at the calyx of Held remains unclear. To provide insight on this, we developed a helper-dependent adenoviral vector (HdAd) that expresses the genetically encoded peroxidase marker for mitochondria, mito-APEX2, at the mouse calyx of Held. We developed protocols to detect labeled mitochondria for use with serial block face scanning electron microscopy to carry out semi-automated segmentation of mitochondria, high-throughput whole terminal reconstruction and presynaptic ultrastructure in mice of either sex. Subsequently, we measured mitochondrial volumes and subsynaptic distributions at the immature postnatal day 7 (P7) and the mature (P21) calyx. We found an increase of mitochondria volumes in terminals and axons from P7 to P21 but did not observe differences between stalk and swelling subcompartments in the mature calyx. Based on these findings, we propose that mitochondrial volumes and synaptic localization developmentally increase to support high firing rates required in the initial stages of auditory information processing.\n\nSignificance StatementElucidating the developmental processes of auditory brainstem presynaptic terminals is critical to understanding auditory information encoding. Additionally, morphological-functional diversity at these terminals is proposed to enhance coding capacity. Mitochondria provide energy for synaptic transmission and can buffer calcium, impacting synaptic plasticity; however, their developmental profile to ultimately support the energetic demands of synapses following the onset of hearing remains unknown. Therefore, we created a helper-dependent adenoviral vector with the mitochondria-targeting peroxidase mito-APEX2 and expressed it at the mouse calyx of Held. Volumetric reconstructions of serial block face electron microscopy data of immature and mature labeled calyces reveal that mitochondrial volumes are increased to support high firing rates upon maturity.

neuroscience↗