TY - JOUR
T1 - Comprehensive analysis of histone post-translational modifications in mouse and human male germ cells
AU - Luense, Lacey J.
AU - Wang, Xiaoshi
AU - Schon, Samantha B.
AU - Weller, Angela H.
AU - Lin Shiao, Enrique
AU - Bryant, Jessica M.
AU - Bartolomei, Marisa S.
AU - Coutifaris, Christos
AU - Garcia, Benjamin A.
AU - Berger, Shelley L.
N1 - Funding Information:
We would like to thank all of the staff at the Penn Andrology Lab for their time and effort in the de-identification of samples used in the study. We would also like to graciously thank K. Karsch for her assistance with spectra interpretation, C. Krapp for technical assistance, and Dr. S. Sidoli and G. Donovan for guidance on statistical analysis. This work was funded by Grants GM055360 (SLB), HD06817 (MSB and SLB), T32HD040135 (SBS), and GM110174 (BAG).
Publisher Copyright:
© 2016 The Author(s).
PY - 2016/6/21
Y1 - 2016/6/21
N2 - Background: During the process of spermatogenesis, male germ cells undergo dramatic chromatin reorganization, whereby most histones are replaced by protamines, as part of the pathway to compact the genome into the small nuclear volume of the sperm head. Remarkably, approximately 90 % (human) to 95 % (mouse) of histones are evicted during the process. An intriguing hypothesis is that post-translational modifications (PTMs) decorating histones play a critical role in epigenetic regulation of spermatogenesis and embryonic development following fertilization. Although a number of specific histone PTMs have been individually studied during spermatogenesis and in mature mouse and human sperm, to date, there is a paucity of comprehensive identification of histone PTMs and their dynamics during this process. Results: Here we report systematic investigation of sperm histone PTMs and their dynamics during spermatogenesis. We utilized "bottom-up" nanoliquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) to identify histone PTMs and to determine their relative abundance in distinct stages of mouse spermatogenesis (meiotic, round spermatids, elongating/condensing spermatids, and mature sperm) and in human sperm. We detected peptides and histone PTMs from all four canonical histones (H2A, H2B, H3, and H4), the linker histone H1, and multiple histone isoforms of H1, H2A, H2B, and H3 in cells from all stages of mouse spermatogenesis and in mouse sperm. We found strong conservation of histone PTMs for histone H3 and H4 between mouse and human sperm; however, little conservation was observed between H1, H2A, and H2B. Importantly, across eight individual normozoospermic human semen samples, little variation was observed in the relative abundance of nearly all histone PTMs. Conclusion: In summary, we report the first comprehensive and unbiased analysis of histone PTMs at multiple time points during mouse spermatogenesis and in mature mouse and human sperm. Furthermore, our results suggest a largely uniform histone PTM signature in sperm from individual humans.
AB - Background: During the process of spermatogenesis, male germ cells undergo dramatic chromatin reorganization, whereby most histones are replaced by protamines, as part of the pathway to compact the genome into the small nuclear volume of the sperm head. Remarkably, approximately 90 % (human) to 95 % (mouse) of histones are evicted during the process. An intriguing hypothesis is that post-translational modifications (PTMs) decorating histones play a critical role in epigenetic regulation of spermatogenesis and embryonic development following fertilization. Although a number of specific histone PTMs have been individually studied during spermatogenesis and in mature mouse and human sperm, to date, there is a paucity of comprehensive identification of histone PTMs and their dynamics during this process. Results: Here we report systematic investigation of sperm histone PTMs and their dynamics during spermatogenesis. We utilized "bottom-up" nanoliquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) to identify histone PTMs and to determine their relative abundance in distinct stages of mouse spermatogenesis (meiotic, round spermatids, elongating/condensing spermatids, and mature sperm) and in human sperm. We detected peptides and histone PTMs from all four canonical histones (H2A, H2B, H3, and H4), the linker histone H1, and multiple histone isoforms of H1, H2A, H2B, and H3 in cells from all stages of mouse spermatogenesis and in mouse sperm. We found strong conservation of histone PTMs for histone H3 and H4 between mouse and human sperm; however, little conservation was observed between H1, H2A, and H2B. Importantly, across eight individual normozoospermic human semen samples, little variation was observed in the relative abundance of nearly all histone PTMs. Conclusion: In summary, we report the first comprehensive and unbiased analysis of histone PTMs at multiple time points during mouse spermatogenesis and in mature mouse and human sperm. Furthermore, our results suggest a largely uniform histone PTM signature in sperm from individual humans.
KW - Epigenetics
KW - Fertility
KW - Histone
KW - Male germ cells
KW - Paternal epigenetics
KW - Post-translational modifications
KW - Sperm
KW - Spermiogenesis
KW - Testes
UR - http://www.scopus.com/inward/record.url?scp=84975883870&partnerID=8YFLogxK
U2 - 10.1186/s13072-016-0072-6
DO - 10.1186/s13072-016-0072-6
M3 - Article
AN - SCOPUS:84975883870
VL - 9
JO - Epigenetics and Chromatin
JF - Epigenetics and Chromatin
SN - 1756-8935
IS - 1
M1 - 24
ER -