Biology subjects

Lackey, E. P.

Publications and source records attributed to Lackey, E. P..

5 recordsLinked to original sources

bioRxiv2025

Structural and functional evidence for ephaptic control of Purkinje cell spike timing by networks of molecular layer interneurons

Axon collaterals of type 1 molecular layer interneurons (MLI1s) contribute to pinceaux that engulf the initial segments of Purkinje cell (PC) axons and generate extracellular signals that ephaptically inhibit PCs. Here we show that a remarkably large number of MLI1s ([~]50) contribute to each pinceau, and that this allows networks of synchronously firing MLI1s to use ephaptic signals to control the precise timing of PC firing in vivo.

neuroscience↗

bioRxiv2025

Purkinje cell collaterals preferentially target a subtype of molecular layer interneuron

In addition to providing outputs from the cerebellar cortex, Purkinje cell (PC) axon collaterals target other PCs, molecular layer interneurons (MLIs), and Purkinje layer interneurons (PLIs). It was recently shown that MLIs consist of two subtypes, but the properties of PC synapses onto these subtypes was not known and it was assumed that all PC collateral to MLI synapses would provide positive feedback to PCs. Clarifying the PC connectivity onto MLI subtypes is vital to understating the influence of feedback from PC collaterals because MLI1s primarily inhibit PCs whereas MLI2s mainly inhibit MLI1s and disinhibit PCs. Here we use a combination of serial EM and optogenetic studies to characterize PC synapses onto MLI subtypes in mice. EM reconstructions show that PCs make 53% of their synapses onto other PCs, 32% onto PLIs, 6% onto MLI1s and 7% onto MLI2s. Since there are far more MLI1s than MLI2s, each MLI2 is expected to receive many more synapses than each MLI1. In slice experiments, optogenetic activation of PCs evokes inhibitory currents in most MLI2s, but primarily disinhibits MLI1s. We also find that candelabrum cells, a type of PLI, form many more synapses onto MLI1s than MLI2s. It is therefore expected that both PC-MLI2-MLI1-PC and PC-PLI-MLI1-PC pathways allow increased PC firing to disinhibit MLI1s, which are known to reduce dendritic PC calcium signals and suppress plasticity at granule cell to PC synapses. These pathways provide negative feedback that act in concert with PC-PC synapses to counter elevations in PC firing. Significance StatementPurkinje cells (PCs) influence processing by inhibiting neurons in the cerebellar cortex, including other PCs, molecular layer interneurons (MLIs) and Purkinje layer interneurons (PLIs). The influence of PC-MLI synapses is not known because there are recently identified MLI subtypes with opposing effects: MLI1s inhibit PCs whereas MLI2s inhibit MLI1s and disinhibit PCs. We used serial EM and optogenetic studies to characterize PC synapses onto MLI subtypes and found that PCs preferentially inhibit MLI2s and disinhibit MLI1s. We also found that candelabrum cells (a type of PLI) preferentially inhibit MLI1s. These findings establish that PC-PC synapses, the PC-MLI2-MLI1-PC pathway and the PC-candelabrum cell-MLI1-PC pathway act together to allow alterations in PC firing to provide negative feedback to other PCs.

neuroscience↗

bioRxiv2020

Maturation of Purkinje cell firing properties relies on granule cell neurogenesis

Preterm infants that suffer cerebellar insults often develop motor disorders and cognitive difficulty. Granule cells are especially vulnerable, and they likely instigate disease by impairing the function of Purkinje cells. Here, we use regional genetic manipulations and in vivo electrophysiology to test whether granule cells help establish the firing properties of Purkinje cells during postnatal mouse development. We generated mice that lack granule cell neurogenesis and tracked the structural and functional consequences on Purkinje cells in these agranular pups. We reveal that Purkinje cells fail to acquire their typical connectivity and morphology, and the formation of characteristic Purkinje cell firing patterns is delayed by one week. We also show that the agranular pups have impaired motor behaviors and vocal skills. These data argue that granule cell neurogenesis sets the maturation time window for Purkinje cell function and refines cerebellar-dependent behaviors.

neuroscience↗

bioRxiv2020

Purkinje cell neurotransmission patterns cerebellar basket cells into zonal modules that are defined by distinct pinceau sizes

Ramon y Cajal proclaimed the neuron doctrine based on circuit features he exemplified using cerebellar basket cell projections. Basket cells form dense inhibitory plexuses that wrap Purkinje cell somata and terminate as pinceaux at the initial segment of axons. Here, we demonstrate that HCN1, Kv1.1, PSD95 and GAD67 unexpectedly mark patterns of basket cell pinceaux that map onto Purkinje cell functional zones. Using cell-specific genetic tracing with an Ascl1CreERT2 mouse conditional allele, we reveal that basket cell zones comprise different sizes of pinceaux. We tested whether Purkinje cells instruct the assembly of inhibitory projections into zones, as they do for excitatory afferents. Genetically silencing Purkinje cell neurotransmission blocks the formation of sharp Purkinje cell zones and disrupts excitatory axon patterning. The distribution of pinceaux into size-specific zones is eliminated without Purkinje cell output. Our data uncover the cellular and molecular diversity of a foundational synapse that revolutionized neuroscience.

neuroscience↗

bioRxiv2018

Molecular layer interneurons shape the spike activity of cerebellar Purkinje cells

One-sentence summaryCerebellar stellate cells and basket cells shape distinct Purkinje cell firing properties\n\nAbstractPurkinje cells receive synaptic input from several classes of interneurons. Here, we address the roles of inhibitory molecular layer interneurons in establishing Purkinje cell function in vivo. Using conditional genetics approaches in mice, we compare how the lack of stellate cell versus basket cell GABAergic neurotransmission sculpts the firing properties of Purkinje cells. We take advantage of an inducible Ascl1CreER allele to spatially and temporally target the deletion of the vesicular GABA transporter, Vgat, in developing neurons. Selective depletion of basket cell GABAergic neurotransmission increases the frequency of Purkinje cell simple spike firing and decreases the frequency of complex spike firing in adult behaving mice. In contrast, lack of stellate cell communication increases the regularity of Purkinje cell simple spike firing while increasing the frequency of complex spike firing. Our data uncover complementary roles for molecular layer interneurons in shaping the rate and pattern of Purkinje cell activity in vivo.

neuroscience↗