In situ cryo-electron tomography reveals the asymmetric architecture of mammalian sperm axonemes

We are grateful to members of the Vale and Agard laboratories for discussions and critical reading of the manuscript. We thank Shixin Yang from the cryo-EM facility at Janelia Research Campus for his assistance with data collection. We thank Caiying Guo for generously sharing the resources required for the mouse experiments. We thank Zanlin Yu, Eric Tse and David Bulkley in the University of California, San Fransisco (UCSF) EM core facility for their assistance with data collection. We thank Sam Li and Shawn Zheng at UCSF for suggestions on EM data processing. We thank Tom Goddard at UCSF for providing a script to mark coordinates of subtomograms. We used and appreciated computing resources at both the workstations at Janelia Research Campus and the HPC Facility at UCSF. Z.C. was supported by the Helen Hay Whitney Foundation Postdoctoral Fellowship. W.M.S. was supported by the National Science Foundation Graduate Research Fellowship Program under grant numbers DGE 1752814 and DGE 2146752. Any opinions, findings and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. P.V.L. received funding from a Pew Biomedical Scholars Award and a GCRLE grant from the Global Consortium for Reproductive Longevity and Equality made possible by the Bia-Echo Foundation. D.A.A. received funding from NIH R35GM118099. R.D.V. received funding from NIH R35GM118106 and the Howard Hughes Medical Institute. The UCSF cryo-EM facility has been supported by NIH grants 1S10OD026881, 1S10OD020054 and 1S10OD021741.

The maps of the following structures are available in the Electron Microscopy Data Bank: EMD-27444, consensus average of 96-nm-repeating structure of mouse doublets; EMD-27445, 32-nm-repeating structure of the central pair complex of mouse sperm; EMD-27446, mouse doublet 1; EMD-27447, mouse doublet 2; EMD-27448, mouse doublet 3; EMD-27449, mouse doublet 4; EMD-27450, mouse doublet 5; EMD-27451, mouse doublet 6; EMD-27452, mouse doublet 7; EMD-27453, mouse doublet 8; EMD-27454, mouse doublet 9; EMD-27455, 48-nm-repeating structure of the doublet microtubule of mouse sperm; EMD-27456, 5–6 bridge of mouse sperm; EMD-27462, consensus average of the 96-nm-repeating structure of human doublets; EMD-27463, 32-nm-repeating structure of the central pair complex of human sperm; EMD-27464, human doublet 1; EMD-27465, human doublet 2; EMD-27466, human doublet 3; EMD-27467, human doublet 4; EMD-27468, human doublet 5; EMD-27469, human doublet 6; EMD-27470, human doublet 7; EMD-27471, human doublet 8; EMD-27473, human doublet 9. The raw tilt series of mouse sperm lamellae and the corresponding tilt angle files are available in the EMPIAR database (EMPIAR-11221).

The script to remap coordinates of subvolumes in three dimensions is provided as Supplementary Code.

The authors declare no competing interests.