Unique Neural Circuit Connectivity of Mouse Proximal, Middle and Distal Colon Defines Regional Colonic Motor Patterns

Abstract Background and Aims Colonic motor patterns have been described by a number of different groups but the neural connectivity and ganglion architecture supporting patterned motor activity have not been elucidated. Our goals were to describe quantitatively, by region, the structural architecture of the mouse ENS and use functional calcium imaging, pharmacology and electrical stimulation to demonstrate regional underpinnings of different motor patterns. Methods Excised colon segments from mice expressing the calcium indicator GCaMP6f or GCaMP6s were used to examine spontaneous and evoked (pharmacological or electrical) changes in GCaMP-mediated fluorescence and coupled with assessment of colonic motor activity, immunohistochemistry, and confocal imaging. 3D image reconstruction and statistical methods were used to describe quantitatively mouse colon myenteric ganglion structure, neural and vascular network patterning and neural connectivity. Results In intact colon, regionally specific myenteric ganglion size, architecture and neural circuit connectivity patterns along with neurotransmitter receptor expression underlie colonic motor patterns that define functional differences along the colon. Region-specific effects on spontaneous, evoked and chemically induced neural activity contribute to regional motor patterns as does intraganglionic functional connectivity. We provide direct evidence of neural circuit structural and functional regional differences that have only been inferred in previous investigations. We include regional comparisons between quantitative measures in mouse and human colon that represent an important advance in demonstrating the usefulness and relevance of the mouse system for translation to the human colon. Conclusions There are several neural mechanisms dependent upon myenteric ganglion architecture and functional connectivity that underlie neurogenic control of pattered motor function in the mouse colon.