Kerbl, A. | bioRxiv Science

bioRxiv2026

Sample preparation optimisation for array tomography volume EM of entire organisms: a case study for marine larvae

Serial-sectioning electron microscopy allows the exploration of the 3D ultrastructure of cells, tissues or whole organisms. Sample preparation is a critical step for this technique, which determines the quality of downstream imaging and analysis. Here we optimize a high-pressure freezing sample-preparation workflow for larvae of the marine annelid Platynereis dumerilii. We tested several fixation and staining methods and screened a large number of specimens by synchrotron X-ray imaging to evaluate specimen integrity and contrast. We further evaluated selected blocks by array tomography SEM to select the best performing protocols. Our protocols and robust workflow could be adapted to similar marine planktonic organisms including ciliated invertebrate larvae.

developmental biology↗

bioRxiv2020

Conservative route to extreme genome compaction in a miniature annelid

The causes and consequences of genome reduction in animals are unclear, because our understanding of this process mostly relies on lineages with often exceptionally high rates of evolution. Here, we decode the compact 73.8 Mb genome of Dimorphilus gyrociliatus, a meiobenthic segmented worm. The D. gyrociliatus genome retains traits classically associated with larger and slower-evolving genomes, such as an ordered, intact Hox cluster, a generally conserved developmental toolkit, and traces of ancestral bilaterian linkage. Unlike some other animals with small genomes, the analysis of the D. gyrociliatus epigenome revealed canonical features of genome regulation, excluding the presence of operons and trans-splicing. Instead, the gene dense D. gyrociliatus genome presents a divergent Myc pathway, a key physiological regulator of growth, proliferation, and genome stability in animals. Altogether, our results uncover a conservative route to genome compaction in annelids, reminiscent of that observed in the vertebrate Takifugu rubripes.

genomics↗

bioRxiv2018

Nerves innervating copulatory organs show common FMRFamide, FVRIamide, MIP and serotonin immunoreactivity patterns across Dinophilidae (Annelida) indicating their conserved role in copulatory behaviour

BackgroundMales of the microscopic annelid family Dinophilidae use their prominent glandomuscular copulatory organ (penis) to enzymatically dissolve the females 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 -tubulin, serotonin, FMRFamide).\n\nResultsSeminal 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.\n\nConclusionsThe 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.

evolutionary biology↗