Single-Nucleus RNA Sequencing of Developing and Mature Superior Colliculus Identifies Neuronal Diversity and Candidate Mediators of Circuit Assembly

Ayupe, Ana C.; Choi, James S.; Beckedorff, Felipe; Mccartan, Robyn; Levay, Konstantin; Park, Kevin K.

doi:10.1101/2023.02.01.526254

Cited by 7 publications

(4 citation statements)

References 60 publications

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“…For example, while GAD2-positive horizontal cells get excitatory inputs from stellate cells (Gale and Murphy, 2018), intracollicular connectivity originating from neighboring excitatory cell types such as narrow-field (NF) and wide-field (WF) cells has shown to be rare (Gale and Murphy, 2018). Next-generation in vitro multi-neuron patch-clamp recordings (Campagnola et al, 2022) could reveal important new insights into the cell-type specific (Ayupe et al, 2023) intracollicular wiring. 4) In addition, collicular neurons in the visual layers also get inputs from deeper SC layers.…”

Section: Discussionmentioning

confidence: 99%

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculusin vivo

Gehr

Sibille

Kremkow

2023

Preprint

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The superior colliculus (SC) is a midbrain structure that contains one of the highest densities of inhibitory neurons in the brain and, together with the thalamocortical visual system, it plays a key role in visually guided behaviors. The SC receives direct inputs from retinal ganglion cells (RGCs) but whether excitatory and inhibitory SC neurons differentially integrate retinal activityin vivois still largely unknown. We recently established an extracellular recording approach using high-density electrodes to measure the activity of RGCs simultaneously with their postsynaptic SC targetsin vivo, that allows addressing how SC neurons integrate RGC activity. Here, we employ this method to study the functional properties and dynamics that govern retinocollicular signaling in a cell-type specific manner by identifying GABAergic SC neurons using optotagging in anesthetized VGAT-ChR2 mice. We measured 305 monosynaptically connected RGC-SC pairs, out of which approximately one third of retinal afferents connect onto inhibitory SC neurons. We show that both excitatory and inhibitory SC neurons receive comparable strong RGC inputs, with functionally similar RGC-SC pairs showing stronger connections. Our results demonstrate that similar wiring rules apply for RGCs innervation of both excitatory and inhibitory SC neurons, which is unlike the cell-type specific connectivity in the thalamocortical system. Contrasting the similar RGC-SC connection strength, we observed that RGC activity contributed more to the activity of postsynaptic excitatory SC neurons than to the activity of postsynaptic inhibitory SC neurons. This implies that the excitatory SC neurons are more specifically coupled to RGC afferent inputs, while inhibitory SC neurons may integrate additional inputs from other sources. Taken together, our study deepens the understanding of cell-type specific retinocollicular functional connectivity and emphasizes that the two major brain areas for visual processing, the visual cortex and the superior colliculus, differently integrate sensory afferent inputs.

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Section: Discussionmentioning

confidence: 99%

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculusin vivo

Gehr

Sibille

Kremkow

2023

Preprint

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show abstract

Section: Discussionmentioning

confidence: 99%

“…They identified five inhibitory clusters of retinorecipient SC cells that allow to specifically map retinocollicular connectivity. In addition, 12 distinct inhibitory subtypes have been recently identified using single-nucleus RNA sequencing where Pax7 was shown to be exclusively expressed in inhibitory neurons (Ayupe et al, 2023). Thus, we are at an enthralling stage where information on SC cell-types finally becomes available based on gene expression patterns, alike to the cortical circuit (Tremblay et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculus in vivo

Gehr

Sibille

Kremkow

2023

eLife

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The superior colliculus (SC) is a midbrain structure that contains one of the highest densities of inhibitory neurons in the brain and, together with the thalamocortical visual system, it plays a key role in visually guided behaviors. The SC receives direct inputs from retinal ganglion cells (RGCs) but whether excitatory and inhibitory SC neurons differentially integrate retinal activity in vivo is still largely unknown. We recently established an extracellular recording approach using high-density electrodes to measure the activity of RGCs simultaneously with their postsynaptic SC targets in vivo, that allows addressing how SC neurons integrate RGC activity. Here, we employ this method to study the functional properties and dynamics that govern retinocollicular signaling in a cell-type specific manner by identifying GABAergic SC neurons using optotagging in anesthetized VGAT-ChR2 mice. We measured 305 monosynaptically connected RGC-SC pairs, out of which approximately one third of retinal afferents connect onto inhibitory SC neurons. We show that both excitatory and inhibitory SC neurons receive comparable strong RGC inputs, with functionally similar RGC-SC pairs showing stronger connections. Our results demonstrate that similar wiring rules apply for RGCs innervation of both excitatory and inhibitory SC neurons, which is unlike the cell-type specific connectivity in the thalamocortical system. Contrasting the similar RGC-SC connection strength, we observed that RGC activity contributed more to the activity of postsynaptic excitatory SC neurons than to the activity of postsynaptic inhibitory SC neurons. This implies that the excitatory SC neurons are more specifically coupled to RGC afferent inputs, while inhibitory SC neurons may integrate additional inputs from other sources. Taken together, our study deepens the understanding of cell-type specific retinocollicular functional connectivity and emphasizes that the two major brain areas for visual processing, the visual cortex and the superior colliculus, differently integrate sensory afferent inputs.

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“…Yet, most information related to SC celltype diversity has been derived based on physiological characteristics of a limited number of experimentally-accessible cells, primarily located in the superficial layers (Gale and Murphy, 2014;Masullo et al, 2019;Shang et al, 2015;Villalobos et al, 2018). Recent efforts have however systematically catalogued cell-types, based on single cell transcriptome, across the embryonic and adult midbrain (Ayupe et al, 2023;La Manno et al, 2021;Xie et al, 2021;Zeisel et al, 2018), and provide therefore an exciting starting point for our general understanding of the extent of SC cell-type diversity.…”

Section: Main Textmentioning

confidence: 99%

Multipotent Progenitors Instruct Ontogeny of the Superior Colliculus

Cheung

Pauler

Koppensteiner

et al. 2023

Preprint

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The superior colliculus (SC) in the mammalian midbrain is essential for multisensory integration, attention, and complex behavior (Basso and May, 2017; Cang et al., 2018). The mature SC cytoarchitecture is organized into distinct laminae and composed of a rich variety of neuronal and glial cell types (Ayupe et al., 2023; Edwards et al., 1986; May, 2006; Xie et al., 2021; Zeisel et al., 2018). Precise execution of the developmental programs regulating the generation of SC cell-type diversity is essential, because deficits due to genetic mutations have been associated with neurodevelopmental diseases and SC dysfunction (Jure, 2018; McFadyen et al., 2020). However, the fundamentals directing the ontogeny of the SC are not well understood. Here we pursued systematic lineage tracing at the single progenitor cell level in order to decipher the principles instructing the generation of cell-type diversity in the SC. We combined in silico lineage reconstruction with a novel genetic MADM (Mosaic Analysis with Double Markers)-CloneSeq approach. MADM-CloneSeq enables the unequivocal delineation of cell lineages in situ, and cell identity based on global transcriptome, of individual clonally-related cells. Our systematic reconstructions of cell lineages revealed that all neuronal cell types in SC emerge from local progenitors without any extrinsic source. Strikingly, individual SC progenitors are exceptionally multipotent with the capacity to produce all known excitatory and inhibitory neuron types of the prospective mature SC, with individual clonal units showing no pre-defined composition. At the molecular level we identified an essential role for PTEN signaling in establishing appropriate proportions of specific inhibitory and excitatory neuron types. Collectively, our findings demonstrate that individual multipotent progenitors generate the full spectrum of excitatory and inhibitory neuron types in the developing SC, providing a novel framework for the emergence of cell-type diversity and thus the ontogeny of the mammalian SC.

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Single-Nucleus RNA Sequencing of Developing and Mature Superior Colliculus Identifies Neuronal Diversity and Candidate Mediators of Circuit Assembly

Cited by 7 publications

References 60 publications

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculusin vivo

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculusin vivo

Retinal input integration in excitatory and inhibitory neurons in the mouse superior colliculus in vivo

Multipotent Progenitors Instruct Ontogeny of the Superior Colliculus

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