Background: Males of the microscopic annelid family Dinophilidae use their prominent glandomuscular copulatory organ (penis) to enzymatically dissolve the female's epidermis and thereafter inject sperm. In order to test for putative conserved copulatory structures and neural orchestration across three dinophilid species, we reconstructed the reproductive myo- and neuroanatomy and mapped neurotransmitter immunoreactivity patterns of two specific markers with reported roles in invertebrate male mating behaviour (FVRIamide, MIP) and three general neural markers (acetylated α-tubulin, serotonin, FMRFamide).
Results: Seminal vesicles (one or two pairs), surrounded by a thin layer of longitudinal and circular muscles and innervated by nerve fibres, are found between testes and copulatory organ in the larger males of Dinophilus vorticoides and Trilobodrilus axi, but are missing in the only 0.05 mm long D. gyrociliatus dwarf males. The midventral copulatory organ is in all species composed of an outer muscular penis sheath and an inner penis cone. Nerves encircle the organ equatorially, either as a ring-shaped circumpenial fibre mass or as dorsal and ventral commissures, which are connected to the ventrolateral nerve cords. All three examined dinophilids show similar serotonin-, FMRFamide-, and FVRIamide-like immunoreactivity patterns in the nerves surrounding the penis, supporting the general involvement of these neurotransmitters in copulatory behaviour in meiofaunal annelids. MIP-like immunoreactivity is restricted to the circumpenial fibre mass in D. gyrociliatus and commissures around the penis in T. axi (but not found in D. vorticoides), indicating its role in controlling the copulatory organ.
Conclusions: The overall myo- and neuroanatomy of the reproductive organs is rather similar in the three studied species, suggesting a common ancestry of the unpaired glandomuscular copulatory organ and its innervation in Dinophilidae. This is furthermore supported by the similar immunoreactivity patterns of the tested neurotransmitters around the penis. Smaller differences in the immunoreactivity patterns around the seminal vesicles and spermioducts might account for additional, individual traits. We thus show morphological support for the putatively conserved role of FMRFamide, FVRIamide, MIP and serotonin in dinophilid copulatory behaviour.
Nerves innervating copulatory organs show common FMRFamide, FVRIamide, MIP and serotonin immunoreactivity patterns across Dinophilidae (Annelida) indicating their conserved role in copulatory behaviour
