Invited Talk ESA-SRB-ANZOS 2025 in conjunction with ENSA

Kingdom of reproductive life; core sperm proteome (124051)

Taylor Pini 1 , Brett Nixon 2 3 , Timothy L Karr 4 5 , Raffaele Teperino 6 7 , Adrián Sanz-Moreno 8 , Patricia da Silva-Buttkus 8 , Frank Tüttelmann 9 , Sabine Kliesch 10 , Valérie Gailus-Durner 8 , Helmut Fuchs 8 , Susan Marschall 6 , Martin Hrabě de Angelis 6 7 8 11 , David Skerrett-Byrne 3 6 7 12
  1. School of Veterinary Science, , The University of Queensland, Gatton, QLD, Australia
  2. Priority Research Centre for Reproductive Science, The University of Newcastle, Newcastle, New South Wales, Australia
  3. Hunter Medical Research Institute, Newcastle, New South Wales, Australia
  4. Biosciences Mass Spectrometry Core Research Facility, Knowledge Enterprise, Arizona State University, Arizona, USA
  5. ASU-Banner Neurodegenerative Disease Research Center, The Biodesign Institute, Arizona, USA
  6. Institute of Experimental Genetics, Helmholtz Zentrum München, Neuherberg, Germany
  7. German Center for Diabetes Research (DZD), Neuherberg, Germany
  8. Institute of Experimental Genetics, German Mouse Clinic, Helmholtz Zentrum München, Neuherberg, Germany
  9. Institute of Reproductive Genetics, Centre of Medical Genetics, University and University Hospital of Münster, Münster, Germany
  10. Department of Clinical and Surgical Andrology, Centre of Reproductive Medicine and Andrology, University and University Hospital of Münster, Münster, Germany
  11. Chair of Experimental Genetics, TUM School of Life Sciences, Technische Universität München, Freising, Germany
  12. School of Biomedical Sciences and Pharmacy, College of Health, Medicine and Wellbeing, The University of Newcastle, Callaghan, NSW, Australia

Reproductive biology is often considered in three siloed research areas; humans, agriculture and wildlife. Yet, each demand solutions for treatment of subfertility, fertility biomarkers, development of assisted reproductive technologies and effective contraception. To efficiently develop solutions applicable to all species, we must improve our understanding of the common biology underpinning reproductive processes. Accordingly, we integrate proteomic data from 29 publicly available datasets (>2 TB of data) to characterize mature sperm proteomes spanning 12 vertebrate species, identifying 13,853 proteins. Although human and mouse have relatively well-annotated sperm proteomes, many non-model species rely heavily on predicted or homology-inferred identifications. Despite variation in proteome size, composition and reproductive strategies, comparative analyses revealed that vertebrates share a fundamental molecular framework essential for sperm function. A core set of 45 species-level and 135 order-level conserved proteins mapped to critical processes, including energy generation, acrosome function, as well as novel signalling pathways (BAG2 and FAT10). Utilising knockout mouse models, we further validate the significance of these conserved proteins, demonstrating that their disruption impairs sperm motility and fertilization capacity. Moreover, we discovered loss-of-function variants of two additional core sperm proteins in clinical samples, linking them to severe sperm defects. Intriguingly, in-silico analysis reveals function-driven, context-dependent diversity surpassing evolutionary patterns. Collectively, these results highlight the value of integrating publicly available datasets and underscore the need for improved genome/proteome annotation in non-model species in mammals. This work provides a foundation for developing cross-species strategies to enhance fertility treatments, assisted reproductive technologies, and conservation efforts. All data is available via ShinySpermKingdom (https://reproproteomics.shinyapps.io/ShinySpermKingdom/).1