Fig. 1

Spermiogenesis in nematodes. a In un-activated spermatids, membranous organelles (shown in teal) migrate to the cell periphery, while Major Sperm Protein (shown in gray) is distributed throughout the cell. Upon sperm activation, Major Sperm Protein forms the pseudopod of the cell and is used to crawl, while the membranous organelles fuse with the cell membrane and release their contents into the extracellular space. b Diagram of the sperm collection processes. Un-activated spermatid proteins were collected by concentrating spermatids collected using microfluidic dissection (see Fig. 2) and lysed to release proteins. For the activated proteome, un-activated spermatids were first collected using a male crushing technique and then concentrated. The supernatant before sperm activation represents a control for cell lysis. Spermatids were activated in vitro by changing the intracellular pH. The supernatant after activation represents the proteins released during membranous organelle fusion. The activated sperm cells were lysed and the membranes pelleted. The supernatant after cell lysis represents the proteins associated with the activated sperm body