PDMS-Based Device for Long-Term Imaging of Gut Peristalsis by Suppressing Tissue Drift in Chicken Embryos

Abstract

Gut peristalsis is essential for the propulsion of luminal contents in the mature gut. Recent studies have suggested that embryonic peristalsis functions as a morphogenetic signal that regulates gut development. For these studies, long-term time-lapse analyses with ex vivo culture must be powerful to understand the dynamics of gut peristalsis. However, a cultured gut specimen undergoes a physical drifting within the microscopic field of view caused by repetitive contractions, preventing long-term quantitative analysis. Here, we developed a simple and inexpensive platform for long-term imaging of embryonic gut peristalsis using a polydimethylsiloxane (PDMS)-based holding device fabricated from a 3D-printed resin mold, which was prepared with a commercially available masked stereolithography printer. The resulting PDMS device successfully held in place a cultured cecum prepared from an embryonic day 10 chicken embryo for 24 h without interfering with intrinsic peristaltic movements and gut elongation. Long-term recordings enabled quantification of the origins, propagation direction, and propagation velocity of peristaltic waves for 24 h. Markedly, these analyses unveiled previously unrecognized differences in peristaltic activity between proximal and distal halves of cecum: while contractions occurred continuously in the proximal half, the distal half exhibited burst and quiescent phases that alternate with an approximately 30-min periodicity. The PDMS platform developed in this study provides a robust and accessible approach for investigating the long-term spatiotemporal dynamics of embryonic gut peristalsis, which will facilitate studies of peristalsis during gut morphogenesis.