Visualizing Spiral Initiation in the Intestine of L. erinacea

The interplay of structure and function is a key concept in biology. My research applies this principle to the spiral intestine found in the skate Leucoraja erinacea, as organ morphology intimately affects organ functionality and organism viability. I am investigating two postulated sources of biological spiraling, mechanical forces and asymmetrical growth, to better understand how the skate intestine gets its distinctive shape. To visualize these factors, I determined the optimal fixation and embedding compounds for two immunofluorescent protocols, 1A4 and Anti-Phospho-Histone-H3. 1A4 is an antibody that detects smooth muscle actin, and can be used to visualize the muscle fibers that may exert constrictive forces during development. Anti-Phospho-Histone-H3 is a cell proliferation marker, which can be used to identify sites of asymmetrical growth within the gut. Both protocols worked best on methanol-fixed, OCT-embedded tissue, showing bright, specific staining. Together, these tools can expand on current understandings of biological spirals and intestinal development. Ultimately, I am interested in this research because of its potential impact on the veterinary and medical fields, as intestinal conditions like volvulus and gastroschisis occur across the animal kingdom. Once we understand the factors that form a healthy gut, we can understand how the process can go wrong and develop therapies to prevent or correct defects.