| Experiment Id | GSE322764 | Name | Lineage and Organ Signals Sequentially Build Organ Intrinsic Nervous Systems [scRNA-seq] |
| Experiment Type | RNA-Seq | Study Type | Baseline |
| Source | GEO | Curation Date | 2026-07-31 |
| description | Organ intrinsic nervous systems (OINSs) are critical components of the bodybrain axis, coordinating visceral organ function with systemic physiological control. Despite their importance, how these distinct neural architectures arise from a common neural crest cell (NCC) origin has remained unclear. Here, we present a systems-level, cross-organ analysis of OINS development, integrating lineage tracing, 3D imaging, single-cell transcriptomics, and genetic perturbations across heart, pancreas, intestine, and lungs. We show that differences in NCC migratory trajectories prefigure the spatial architecture of OINSs, laying the foundation for organ-specific patterning. In contrast, molecular identity emerges largely in response to local environments, indicating that extrinsic cues play a major instructive role. Using in vitro co-cultures, we demonstrate that organ-derived cues reprogram intrinsic neurons toward organ-specific transcriptional profiles and direct neuronal differentiation, with extracellular matrix (ECM) contact identified as a central mediator. In vivo, ECMintegrin signaling supports intrinsic cardiac neuron neurogenesis, while ECM crosslinking stabilizes their stereotyped ganglionic organization. Together, these findings reveal that OINS diversity arises through a dual logic: lineage programs prefigure spatial frameworks, while organ-specific cues instruct final molecular identities and architectural precision. This work establishes a conceptual paradigm for how organs actively build their own nervous systems, illuminating principles that underpin bodybrain integration. For single-cell RNA sequencing of organ-intrinsic neuronal cells, the heart, lungs, pancreas and intestines were dissected from Wnt1-Cre;lox-tdTomato, Phox2b-Flpo;frt-tdTomato, or Baf53b-Cre;lox-L10GFP mouse embryos at E12.5, E14.5, E16.5, E18.5 and P56. Fluorescently labelled neuronal cells were enriched by Fluorescence-activated cell sorting (FACS) on a BD FACS Aria cell sorter. For experiments comparing heart near and heart far regions, E14.5 atrial regions containing Wnt1-Cre;lox-L10GFP labelled ICNS cells were separated from atrial regions lacking ICNS. Each region was dissociated into single-cell suspensions. For co-culture experiments, cardiac atria and intestines were dissected from E13.5 Phox2b-Flpo;frt-tdTomato embryos. FACS-purified enteric neurons were co-cultured with non-neuronal cardiac or intestinal cells for 4-5 days before harvesting for single-cell RNA sequencing. Single-cell cDNA libraries were prepared using the Chromium Single Cell 3 V3 reagent kit and sequenced using an Illumina NovaSeq 6000 sequencer. |
