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Heckscher, E.

Publications and source records attributed to Heckscher, E..

6 recordsLinked to original sources

Multiple scales of coordination along the body axis during Drosophila larval locomotion

Coordinated movement along the body axis is critical to locomotion. In segmented, limbless animals, anterior (head) and posterior (tail) segments play different roles in locomotion, leading to a need for flexible coordination across body regions. Larval Drosophila melanogaster present a tractable experimental model for limbless, segmented crawling given the extensive genetic tools available and the optical clarity of the body. Prior work has suggested that, during crawling, all larval body segments contract similarly, despite the fact that each crawl cycle comprises two overlapping phases: a piston involving the most posterior segments, and a peristaltic wave involving all body segments. To test whether coordination varies regionally during locomotion, we expressed GCaMP in body wall muscles of larvae of either sex, and recorded segmental contraction kinematics and muscle recruitment during many cycles of locomotion in linear channels. Facilitated by machine vision techniques, we discovered new features of larval crawling at multiple scales. First, the propagation of both contraction and recruitment waves slowed approaching mid-body segments, then sped up towards the head. Second, the timing relationship between contraction and recruitment waves could be highly variable in anterior segments. Third, contraction durations showed particularly strong intersegmental correlations among posterior segments. These data suggest posterior segments coordinate to power the piston phase while anterior segments tolerate greater flexibility to enable reorienting behaviors. Our results depict an unanticipated degree of axial heterogeneity in the coordination of limbless crawling, opening new avenues to study the origins of whole-body coordination and the consequences of segmental diversity for locomotion. SIGNIFICANCE STATEMENTMost animals move through the world by coordinating muscle contractions across the bodys many segments. Studying how segments are coordinated in limbless animals can uncover principles that are fundamental for movement across the animal kingdom. However, in one of the best genetic model organisms to address this question, the larval fruit fly, it was unknown how the nearly identical body segments are coordinated across the body. We used machine vision techniques to discover an unanticipated degree of heterogeneity: the timings of muscle activity and movements are more coordinated among tail-end than head-end segments. This provides new insights into animal movements, and opens opportunities to study how interactions of the body and nervous system coordinate segmental movements across an entire animal.

neuroscience↗

Temporal cohort number, neural composition, and connectivity vary along the Drosophila larval body axis

Neural stem cells are inherently flexible, producing different lineages of neurons in different contexts; yet in vivo, development must be highly regulated because aberrant stem cell development leads to microcephalies, tumors, and neurodevelopmental disorders. However, the full complement of in vivo flexibility in neural stem cell development remains poorly understood. Within the Drosophila CNS, 30 types of neural stem cells or neuroblasts repeat in each segment with segment-specific differences in development. We studied NB3-3 neuroblast in each of the 14 segments where it is present. In different segments, NB3-3 produces different types of neurons in different numbers but always produces Even-skipped Lateral (EL) neurons. Prior work showed that in abdominal segments, ELs comprise two types of temporal cohorts. Temporal cohorts are small sets of neurons within a lineage that are born within a tight time window. In the abdomen, EL neurons in the early-born temporal cohort all receive mechanosensory input, whereas EL neurons in the late-born temporal cohort all receive proprioceptive input. Herein, we find that NB3-3 generates an early-born EL temporal cohort in all segments. In specific regions, NB3-3 neuroblasts produce additional types of temporal cohorts, including but not limited to the late-born EL temporal cohort. We show that variations in NB3-3 division pattern determine the number and type of temporal cohorts. Later in development, the number of neurons within a temporal cohort is refined by a variety of post-mitotic mechanisms: regulation of molecular identity markers, cell death, and differentiation status, acting in complex combinations in different segments. Post-mitotic refinement of neuron numbers occurs most often for neurons of the early-born EL temporal cohort. Further, neurons of the early-born EL temporal cohort are incorporated into different circuits, depending on segment. Together, our results show how a single neural stem cell type can flexibly generate segment-specific lineages and contribute to various circuits. We offer the following framework for understanding how a given type of neural stem cell generates diverse temporal cohorts: a stem cell type invariably generates one type of temporal cohort, regardless of location or pattern of division. For neurons in this invariant temporal cohort, post-mitotic mechanisms introduce diversity in number. The stem cell, on a segment-by-segment basis, can also produce additional types of temporal cohorts simply by altering its division pattern. This framework is valuable for motor circuit development, the development of sexual dimorphisms, nervous system evolution, and in vitro stem cell applications.

neuroscience↗

Temporal transcription factors determine circuit membership by altering motor neuron-to-muscle synaptic partnerships

Previously, using the Drosophila motor system as a model, we found the classic temporal transcription factor, Hunchback acts in NB7-1 neuronal stem cells as a molecular switch to control which circuits are populated by NB7-1 neuronal progeny (Meng et al., 2019). Here, we manipulate cardinal transcription factors, Nkx6 and Hb9, which are candidate effectors of Hunchback and which alter axon pathfinding in embryos. Yet manipulation of these cardinal transcription factors does not permanently alter neuromuscular synaptic partnerships. This demonstrates that compensation can correct early defects. We perform additional temporal transcription factor manipulations, precociously expressing Pdm and Castor in NB7-1 and prolonging expression of Hunchback in NB3-1. In every case, we find permanent alterations in neuromuscular synaptic partnerships. These data support the idea that temporal transcription factors are uniquely potent determinants of circuit membership, which do not trigger compensatory programs because they act to establish the expected pattern of wiring for the motor system.

developmental biology↗

RNA-binding protein Syncrip regulates Starvation-Induced Hyperactivity in adult Drosophila

How to respond to starvation determines fitness. One prominent behavioral response is increased locomotor activities upon starvation, also known as Starvation-Induced Hyperactivity (SIH). SIH is paradoxical as it promotes food seeking but also increases energy expenditure. Despite its importance in fitness, the genetic contributions to SIH as a behavioral trait remains unexplored. Here, we examined SIH in the Drosophila melanogaster Genetic Reference Panel (DGRP) and performed genome-wide association studies. We identified 23 significant loci, corresponding to 14 genes, significantly associated with SIH in adult Drosophila. Gene enrichment analyses indicated that genes encoding ion channels and mRNA binding proteins (RBPs) were most enriched in SIH. We are especially interested in RBPs because they provide a potential mechanism to quickly change protein expression in response to environmental challenges. Using RNA interference, we validated the role of syp in regulating SIH. syp encodes Syncrip (Syp), an RBP. While ubiquitous knockdown of syp led to semi-lethality in adult flies, adult flies with neuron-specific syp knockdown were viable and exhibited decreased SIH. Using the Temporal and Regional Gene Expression Targeting (TARGET) system, we further confirmed the role of Syp in adult neurons in regulating SIH. To determine how syp is regulated by starvation, we performed RNA-seq using the heads of flies maintained under either food or starvation conditions. RNA-seq analyses revealed that syp was alternatively spliced under starvation while its expression level was unchanged. We further generated an alternatively-spliced-exon-specific knockout (KO) line and found that KO flies showed reduced SIH. Together, this study demonstrates a significant genetic contribution to SIH as a behavioral trait, identifies syp as a SIH gene, and highlights the significance of RBPs and post-transcriptional processes in the brain in regulating behavioral responses to starvation. Author summaryAnimals living in the wild often face periods of starvation. How to physiologically and behaviorally respond to starvation is essential for survival. One behavioral response is Starvation-Induced Hyperactivity (SIH). We used the Drosophila melanogaster Genetic Reference Panel, derived from a wild population, to study the genetic basis of SIH. Our results show that there is a significant genetic contribution to SIH in this population, and that genes encoding RNA binding proteins (RBPs) are especially important. Using RNA interference and the TARGET system, we confirmed the role of an RBP Syp in adult neurons in SIH. Using RNA-seq and Western blotting, we found that syp was alternatively spliced under starvation while its expression level was unchanged. Further studies from syp exon-specific knockout flies showed that alternative splicing involving two exons in syp was important for SIH. Together, this study identifies syp as a SIH gene and highlights an essential role of post-transcriptional modification in regulating this behavior.

genomics↗

How prolonged expression of Hunchback, a temporal transcription factor, re-wires locomotor circuits

AbstractIn many CNS regions, neuronal birth timing is associated with circuit membership. In Drosophila larvae, we show U motor neurons are a temporal cohort--a set of non-identical, contiguously-born neurons from a single neuronal stem cell that contribute to the same circuit. We prolong expression of a temporal transcription factor, Hunchback, to increase the number of U motor neurons with early-born molecular identities. On the circuit level, this expands and re-wires the U motor neuron temporal cohort. On the cell biological level, we find novel roles for Hunchback in motor neuron target selection, neuromuscular synapse formation, dendrite morphogenesis, and behavior. These data provide insight into the relationship between stem cell and circuit, show that Hunchback is a potent regulator of circuit assembly, and suggest that temporal transcription factors are molecules that could be altered during evolution or biomedical intervention for the generation of novel circuits.

developmental biology↗

Proprioceptive neurons of larval Drosophila melanogaster show direction selective activity requiring the mechanosensory channel TMC.

Drosophila Transmembrane channel-like (Tmc) is a protein that functions in larval proprioception. The closely related TMC1 protein is required for mammalian hearing, and is a pore forming subunit of the hair cell mechanotransduction channel. In hair cells, TMC1 is gated by small deflections of microvilli that produce tension on extracellular tip-links that connect adjacent villi. How Tmc might be gated in larval proprioceptors, which are neurons having a morphology that is completely distinct from hair cells, is unknown. Here, we have used high-speed confocal microscopy to both measure displacements of proprioceptive sensory dendrites during larval movement, and to optically measure neural activity of the moving proprioceptors. Unexpectedly, the pattern of dendrite deformation for distinct neurons was unique and differed depending on the direction of locomotion: ddaE neuron dendrites were strongly curved by forward locomotion while the dendrites of ddaD were more strongly deformed by backward locomotion. Furthermore, GCaMP6f calcium signals recorded in the proprioceptive neurons during locomotion indicated tuning to the direction of movement. ddaE showed strong activation during forward locomotion while ddaD showed responses that were strongest during backwards locomotion. Peripheral proprioceptive neurons in animals mutant for Tmc showed a near complete loss of movement related calcium signals. As the strength of the responses of wild type animals was correlated with dendrite curvature, we propose that Tmc channels may be activated by membrane curvature in dendrites that are exposed to strain. Our findings begin to explain how distinct cellular systems rely on a common molecular pathway for mechanosensory responses.

neuroscience↗