o:3466 Epigenetic inheritance of diet-induced and sperm-borne mitochondrial RNAs en Spermatozoa harbour a complex and environment-sensitive pool of small non-coding RNAs (sncRNAs)1, which influences offspring development and adult phenotypes1-7. Whether spermatozoa in the epididymis are directly susceptible to environmental cues is not fully understood8. Here we used two distinct paradigms of preconception acute high-fat diet to dissect epididymal versus testicular contributions to the sperm sncRNA pool and offspring health. We show that epididymal spermatozoa, but not developing germ cells, are sensitive to the environment and identify mitochondrial tRNAs (mt-tRNAs) and their fragments (mt-tsRNAs) as sperm-borne factors. In humans, mt-tsRNAs in spermatozoa correlate with body mass index, and paternal overweight at conception doubles offspring obesity risk and compromises metabolic health. Sperm sncRNA sequencing of mice mutant for genes involved in mitochondrial function, and metabolic phenotyping of their wild-type offspring, suggest that the upregulation of mt-tsRNAs is downstream of mitochondrial dysfunction. Single-embryo transcriptomics of genetically hybrid two-cell embryos demonstrated sperm-to-oocyte transfer of mt-tRNAs at fertilization and suggested their involvement in the control of early-embryo transcription. Our study supports the importance of paternal health at conception for offspring metabolism, shows that mt-tRNAs are diet-induced and sperm-borne and demonstrates, in a physiological setting, father-to-offspring transfer of sperm mitochondrial RNAs at fertilization. Energy Metabolism; Epigenetics; Animals; Male; Spermatozoametabolism; Mice; RNA; Mitochondrial Genetics Metabolism; Female; Epigenesis, Genetic; 1552099 10.1038/s41586-024-07472-3 2024-09-02T08:39:19.275Z 44 yes 46 A. Tomar Helmholtz Zentrum München / German Center for Diabetes Research M. Gomez-Velazquez Helmholtz Zentrum München / German Center for Diabetes Research person R. Gerlini Helmholtz Zentrum München / German Center for Diabetes Research person G. Comas-Armangué Helmholtz Zentrum München / German Center for Diabetes Research person L. Makharadze Helmholtz Zentrum München / German Center for Diabetes Research person Thomas Kolbe University of Veterinary Medicine Vienna / BOKU University person 0000-0003-2974-8066 Auke Boersma University of Veterinary Medicine Vienna person 0000-0002-3066-2836 Mark Dahlhoff University of Veterinary Medicine Vienna person 0000-0001-9189-7631 Jörg Burgstaller University of Veterinary Medicine Vienna / Boku University person M. Lassi Helmholtz Zentrum München / German Center for Diabetes Research person J. Darr Helmholtz Zentrum München / German Center for Diabetes Research person J. Toppari University of Turku person H. Virtanen University of Turku person A. Kühnapfel Leipzig University person 0000-0003-3668-0784 M. Scholz Leipzig University person 0000-0002-4059-1779 K. Landgraf Leipzig University person 0000-0002-6878-6033 W. Kiess Leipzig University person M. Vogel Leipzig University person V. Gailus-Durner Helmholtz Zentrum München / German Center for Diabetes Research person 0000-0002-6076-0111 H. Fuchs Helmholtz Zentrum München / German Center for Diabetes Research person S. Marschall Helmholtz Zentrum München / German Center for Diabetes Research person 0000-0001-5191-7060 M. Hrabě de Angelis Helmholtz Zentrum München / German Center for Diabetes Research person N. Kotaja University of Turku person A. Körner Leipzig University person 0000-0001-6001-0356 R. Teperino Helmholtz Zentrum München / German Center for Diabetes Research person 0000-0001-8815-1409 application/pdf 12407293 https://phaidra.vetmeduni.ac.at/o:3466 no yes 16 70 1552253 Nature 27 630 8017 720 727 Nature Portfolio 2024