Visualization of enteric neural crest cell migration in SOX10 transgenic mouse gut using time-lapse fluorescence imaging

BackgroundEnteric neural crest cells (ENCCs) were labeled with VENUS, an enhanced green fluoroscein protein, to record their migration in genetically engineered transgenic (SOX10-VENUS) mice.Materials and MethodsPregnant SOX10-VENUS mice were killed on day 12.5 of gestation. The colorectum was excised from each embryo (n = 20) and placed in tissue culture medium. Time-lapse images captured using fluorescence microscopy at 10-minute intervals for 3000 minutes were compiled into a video to display ENCC migration.ResultsAt 0 minutes, VENUS+ ENCC were observed to be clustered in the cecum and proximal colon (vagal ENCC), and similar cells were also seen in the rectum/sacrum (sacral ENCC). After 500 minutes, vagal VENUS+ ENCC had migrated caudally from the proximal colon to the midcolon, reaching the distal colon after 800 minutes. Sacral VENUS+ ENCC had migrated rostrally and transversely by 1250 minutes and had integrated with vagal ENCC by 2500 minutes.ConclusionWe recorded the actual rostral-to-caudal migration of vagal ENCC, caudal-to-rostral migration of sacral ENCC, and their integration in the developing mouse hindgut. Such direct evidence of ENCC migration may further elucidate understanding of ENCC development, thus providing insight into the histopathology of bowel dysmotility disorders.