Axon Fasciculation, Mediated by Transmembrane Semaphorins, Is Critical for the Establishment of Segmental Specificity of Corticospinal Circuits.

Axon fasciculation is thought to be a critical step in neural circuit formation and function. Recent studies have revealed various molecular mechanisms that underlie axon fasciculation; however, the impacts of axon fasciculation, and its corollary, defasciculation, on neural circuit wiring remain unclear. Corticospinal (CS) neurons in the sensorimotor cortex project axons to the spinal cord to control skilled movements. In rodents, the axons remain tightly fasciculated in the brain and traverse the dorsal funiculus of the spinal cord. Here we show that plexinA1 (PlexA1) and plexinA3 (PlexA3) receptors are expressed by CS neurons, whereas their ligands, semaphorin-5A (Sema5A) and semaphorin-5B (Sema5B) are expressed in the medulla at the decussation site of CS axons to inhibit premature defasciculation of these axons. In the absence of Sema5A/5B-PlexA1/A3 signaling, some CS axons are prematurely defasciculated in the medulla of the brainstem, and those defasciculated CS axons aberrantly transverse in the spinal gray matter instead of the spinal dorsal funiculus. In the absence of Sema5A/Sema5B-PlexA1/A3 signaling, CS axons, which would normally innervate the lumbar spinal cord, are unbundled in the spinal gray matter, and prematurely innervate the cervical gray matter with reduced innervation of the lumbar gray matter. In both Sema5A/5B and PlexA1/A3 mutant mice (both sexes), stimulation of the hindlimb motor cortex aberrantly evokes robust forelimb muscle activation. Finally, Sema5A/5B and PlexA1/A3 mutant mice show deficits in skilled movements. These results suggest that proper fasciculation of CS axons is required for appropriate neural circuit wiring and ultimately affect the ability to perform skilled movements.SIGNIFICANCE STATEMENT Axon fasciculation is believed to be essential for neural circuit formation and function. However, whether and how defects in axon fasciculation affect the formation and function of neural circuits remain unclear. Here we examine whether the transmembrane proteins semaphorin-5A (Sema5A) and semaphorin-5B (Sema5B), and their receptors, plexinA1 (PlexA1) and plexinA3 (PlexA3) play roles in the development of corticospinal circuits. We find that Sema5A/Sema5B and PlexA1/A3 are required for proper axon fasciculation of corticospinal neurons. Furthermore, Sema5A/5B and PlexA1/A3 mutant mice show marked deficits in skilled motor behaviors. Therefore, these results strongly suggest that proper corticospinal axon fasciculation is required for the appropriate formation and functioning of corticospinal circuits in mice.

Independent premotor encoding of the sequence and structure of birdsong in avian cortex.

How the brain coordinates rapid sequences of learned behavior, such as human speech, remains a fundamental problem in neuroscience. Birdsong is a model of such behavior, which is learned and controlled by a neural circuit that spans avian cortex, basal ganglia, and thalamus. The songs of adult male zebra finches (Taeniopygia guttata), produced as rapid sequences of vocal gestures (syllables), are encoded by the cortical premotor region HVC (proper name). While the motor encoding of song within HVC has traditionally been viewed as unitary and distributed, we used an ablation technique to ask whether the sequence and structure of song are processed independently within HVC. Results revealed a functional topography across the medial-lateral axis of HVC. Bilateral ablation of medial HVC induced a positive disruption of song (increase in atypical syllable sequences), whereas bilateral ablation of lateral HVC induced a negative disruption (omission of individual syllables). Bilateral ablation of central HVC either had no effect on song or induced syllable omission, similar to lateral HVC ablation. We then investigated HVC connectivity and found parallel afferent and efferent pathways that transit medial and lateral HVC and converge at vocal motor cortex. In light of recent evidence that syntactic and lexical components of human speech are processed independently by neighboring regions of cortex (Menenti et al., 2012), our demonstration of anatomically distinct pathways that differentially process the sequence and structure of birdsong in parallel suggests that the vertebrate brain relies on a common approach to encode rapid sequences of vocal gestures.

Axial organization of a brain region that sequences a learned pattern of behavior.

Neural activity within HVC (proper name), a premotor nucleus of the songbird telencephalon analogous to premotor cortical regions in mammals, controls the temporal structure of learned song in male zebra finches (Taeniopygia guttata). HVC is composed of a superficially isomorphic neuronal mosaic, implying that song is encoded in a distributed network within HVC. Here, we combined HVC microlesions (10% focal ablation) with singing-driven immediate-early gene (IEG) labeling to explore the network architecture of HVC during singing. Microlesions produce a transient disruption of HVC activity that results in a temporary (≈ 1 week) loss of vocal patterning. Results showed an asymmetrical reduction in the density of IEG-labeled cells 3-5 d after microlesions: swaths of unlabeled cells extended rostrally and/or caudally depending on the position of the HVC microlesion. Labeling returned once birds recovered their songs. Axial swaths of unlabeled cells occurred whether microlesions were located at rostral or caudal poles of HVC, indicating that the localized reduction in IEG labeling could not be attributable solely to transection of afferents that enter HVC rostrally. The asymmetrical pattern of reduced IEG labeling could be explained if synaptic connectivity within HVC is organized preferentially within the rostrocaudal axis. In vivo retrograde tracer injections and in vitro stimulation and recording experiments in horizontal slices of HVC confirmed a rostrocaudal organization of HVC neural connectivity. Our findings suggest that HVC contains an axially organized network architecture that may encode the temporal structure of song.

Dual pre-motor contribution to songbird syllable variation.

Many forms of learning, including songbird vocal learning, rely on the brain's ability to use pre-motor variation and sensory feedback to guide behavior toward a specific target or goal. In the vocal control system of zebra finches (Taeniopygia guttata) the pre-motor mechanisms of vocal variation are thought to be vested primarily in a neural pathway that includes the basal ganglia. A second circuit that includes avian analogues of mammalian pre-motor and motor cortex (the vocal motor pathway) generates the patterned structure of learned adult song. Here, we tested the ability of the basal ganglia pathway to generate pre-motor vocal variation within the spectral and temporal dimensions of zebra finch song structure. In adult birds, ablation of the basal ganglia pathway significantly reduced the spectral and temporal dispersion of individual song syllables, with the exception of syllable pitch, where the reduction was not statistically significant when compared against surgical controls. We found a similar pattern of results using longitudinal comparisons (juvenile vs adult) to isolate the contribution of the basal ganglia pathway to spectral dispersion in populations of developing song syllables--variation in syllable pitch was significantly smaller than in all other measured spectral features. The results indicate that pre-motor variation generated by the basal ganglia pathway may be sufficient to adjust vocal output toward highly acoustically dispersed targets of imitation, but suggest that complete acquisition of the pronounced variation in syllable pitch that characterizes adult song will necessitate a gradual developmental interaction between the basal ganglia and vocal motor pathways.

Corticospinal Pathways and Interactions Underpinning Dexterous Forelimb Movement of the Rodent.

In 2013, Thomas Jessell published a paper with Andrew Miri and Eiman Azim that took on the task of examining corticospinal neuron function during movement (Miri et al., 2013). They took the view that a combination of approaches would be able to shed light on corticospinal function, and that this function must be considered in the context of corticospinal connectivity with spinal circuits. In this review, we will highlight recent developments in this area, along with new information regarding inputs and cross-connectivity of the corticospinal circuit with other circuits across the rodent central nervous system. The genetic and viral manipulations available in these animals have led to new insights into descending circuit interaction and function. As species differences exist in the circuitry profile that contributes to dexterous forelimb movements (Lemon, 2008; Yoshida and Isa, 2018), highlighting important advances in one model could help to compare and contrast with what is known about other models. We will focus on the circuitry underpinning dexterous forelimb movements, including some recent developments from systems besides the corticospinal tract, to build a more holistic understanding of sensorimotor circuits and their control of voluntary movement. The rodent corticospinal system is thus a central point of reference in this review, but not the only focus.