Spermatozoa of Sminthopsis murina (Mammalia: Metatheria) exhibit an unusually high degree of chromatin stability in the absence of disulphide bonding in protamine 1
S. D. Johnston A C , C. López-Fernández B , F. Arroyo B , S. Fardell A , R. Roy B and J. Gosálvez B
+ Author Affiliations
- Author Affiliations
A School of Agriculture and Food Science, The University of Queensland, Gatton, Qld 4343, Australia.
B Unit of Genetics, Department of Biology, Universidad Autónoma de Madrid, 20849 Madrid, Spain.
C Corresponding author. Email: s.johnston1@uq.edu.au
Reproduction, Fertility and Development 28(9) 1268-1275 https://doi.org/10.1071/RD14504
Submitted: 12 September 2014 Accepted: 5 January 2015 Published: 25 February 2015
Abstract
Although all but a single genus (Planigale) of the metatheria so far examined contain no cysteine residues in protamine 1, we report a remarkable level of chromatin stability in the spermatozoa of the common dunnart, Sminthopsis murina. S. murina cauda epididymal spermatozoa and somatic epithelial cells were exposed to a combination of graded treatments to lyse sperm protein and induce sperm DNA damage via standard freeze–thaw protocols and post-thaw incubation at 37°C for 48 h, exposure to sodium nitroprusside (SNP) and the enzyme AluI restriction endonuclease. Sperm DNA fragmentation was assessed using the comet assay and sperm chromatin dispersal test. Although S. murina somatic cells showed DNA fragmentation following protein lysis and after treatment with all the protocols specifically designed to induce chromatin damage, sperm DNA fragmentation was only observed following moderate to severe proteolytic exposure and treatment with the restriction endonuclease; there was also an increase in the baseline halo of spermatozoa treated with an aggressive reducing agent, but no corresponding evidence of fragmented DNA, suggesting that cysteine residues may be functioning to conform tertiary and/or quaternary chromatin structure. Given that the protamine 1 of S. murina contains no cysteine, we suggest that the source of these residues is possibly the histone fraction of the chromatin and that the high level of stability is potentially related to prolonged sperm survival in the female’s reproductive tract.
Additional keywords: cysteine residues, Halomax, marsupial, sperm chromatin dispersion assay, sperm DNA fragmentation.
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