Mutation of <i>SPSL1</i> leads to defects in spermatophore formation, sperm distribution, and sperm activation.

(A, A’) Representative images of the copulatory bursa of virgin WT female. Scale bars, 1mm. (B, B’) Representative images of copulatory bursa of WT females mated with WT or SPSL1-mutant males 1 hour-post copulation (hpc). Scale bars, 1mm. (C, C’) Representative images of spermatophore dissected from the copulatory bursa of females mated WT or SPSL1-mutant males 1 hpc. Scale bars, 1 mm. (D, D’) Schematic of the spermatophore inside in the copulatory bursa of WT females mated with WT or SPSL1-mutant males. scale bars, 1mm. (E) Representative images of sperm bundles released from spermatophores from indicated mating at 1 hpc. The green arrows indicate eupyrene sperm bundles, magenta arrows indicate apyrene sperm. Blue, Hoechst. Scale bars, 50 μm. (F) The frequencies of eupyrene sperm bundles (ESB), eupyrene sperm bundles in dissociation (ESB-D), eupyrene sperm (ES) released from spermatophores. (G) Representative images of sperm bundles released from spermatophore-excluded copulatory bursae at 1 hpc for indicated matings. The green arrows indicate eupyrene sperm bundles. Blue, Hoechst. Scale bars, 50 μm. (H) The frequencies of ESB, ESB-D, ES released from spermatophore-excluded copulatory bursae. (I-K) The numbers of I) ESB, J) ESB-D, K) ES per field of view released from copulatory bursae at 1 hpc for indicated matings. For F and H, 110 fields of view from 22 females mated with WT males and 100 fields of view from 20 females mated with SPSL1-mutant males were analyzed. For I-K, 30 fields of view from 6 females for indicated matings were analyzed. Data are means ± SEM (*p p p t-test). ΔSPSL1 represents SPSL1 mutants.