Sedimentation properties in density gradients correspond with levels of sperm DNA fragmentation, chromatin compaction and binding affinity to hyaluronic acid

Abstract

ReuseUnless indicated otherwise, fulltext items are protected by copyright with all rights reserved. The copyright exception in section 29 of the Copyright, Designs and Patents Act 1988 allows the making of a single copy solely for the purpose of non-commercial research or private study within the limits of fair dealing. The publisher or other rights-holder may allow further reproduction and re-use of this version -refer to the White Rose Research Online record for this item. Where records identify the publisher as the copyright holder, users can verify any specific terms of use on the publisher's website. TakedownIf you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing eprints@whiterose.ac.uk including the URL of the record and the reason for the withdrawal request. Accepted ManuscriptTitle: Sedimentation properties in density gradients correspond with levels of sperm DNA fragmentation, chromatin compaction and binding affinity to hyaluronic acid 22-11-2016 Please cite this article as: Forough Torabi, Adel Binduraihem, David Miller, Sedimentation properties in density gradients correspond with levels of sperm DNA fragmentation, chromatin compaction and binding affinity to hyaluronic acid, Reproductive BioMedicine Online (2016), http://dx.doi.org/doi: 10.1016/j.rbmo.2016.11.011. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractMature spermatozoa bind hyaluronic acid in the extracellular matrix via hyaladherins.Immature spermatozoa may be unable to interact because they do not express the appropriate hyaladherins on their surface. Fresh human semen samples were fractionated using differential density gradient centrifugation (DDGC) and the ability of these fractions to bind hyaluronic acid was evaluated. The presence of sperm hyaladherins was also assessed. CD44 was located mainly on the acrosome and equatorial segment and became more restricted to the equatorial segment in capacitated spermatozoa. Hyaluronic acid-TRITC (hyaluronan acid conjugated with tetramethylrhodamine isothiocyanante), a generic hyaluronic-acid-binding reagent, labelled the membrane and the neck region, particularly after capacitation. Sperm populations obtained after DDGC or after interaction with hyaluronic acid were assessed for DNA fragmentation and chromatin maturity. Strong relationships between both measures and sperm sedimentation and hyaluronic-acid-binding profiles were revealed. Capacitation Overall success rates for assisted conception procedures, however (including ICSI), have remained relatively static, with about 24% of cycles for all age groups deliverin...

Sperm selection for assisted reproduction by prior hyaluronan binding: the HABSelect RCT

“…These and other reports have been linked to low sperm maturity in relation to high cytoplasmic retention, a feature of sperm abnormality. 46,123,124 AB was also shown to discriminate between sperm isolated from native semen (mixed compaction) and sperm pelleted following DGC (good compaction), similar to the processed sperm used in HABSelect 125 (and see Sakkas et al 43 and Torabi et al 104 ). In relation to achieving a CP, compaction defects, when relevant, seemed not to be offset by PICSI, which offered no advantage.…”

“…Quid pro quo, HABSelect's data appear to confirm the efficacy of semen washing by differential DGC. 43,104,129 The data also inform the debate on the utility of measuring sperm DNA fragmentation in the clinical context, in which disagreements are common and the issue of the relative merits of different assays as well as their differing reporting criteria are all contentious. 36,83,[130][131][132] The American Society for Reproductive Medicine's 2013 position paper was generally not supportive of their introduction to the clinic, 133 but others argued that this recommendation was short-sighted 134 or took a softer position, indicating that further research was needed.…”

“…The binding conditions help to ensure that only motile sperm with high vitality are used for ICSI and the available research data on the properties of HA-binding sperm support this modus operandi. 49,[104][105][106][107] HABSelect had to make use of a UK-and MHRA-registered product/procedure in the clinical context, and although solid or liquid-based products could have been used, PICSI was chosen because it was thought to be more physiologically relevant to the natural sperm-HA solid-state interaction that takes place in the female reproductive tract and because it was more straightforward to train operatives in its use to a uniform standard among multiple clinics. PICSI had also been more widely reported in the context of CPR and miscarriage.…”

“…Considering that sperm were being directly injected into eggs, it is likely that the main arbiter of successful fertilisation was gamete quality, which HBS has been shown to predict for sperm relative to their concentration and GENERAL DISCUSSION AND CONCLUSIONS NIHR Journals Library www.journalslibrary.nihr.ac.uk motility. 104,106,108 There are disagreements, however, on the assay's ability to screen for improved clinical outcomes. 117 Fertilisation rates for the trial cohort were similar to the rates reported by Majumdar and Majumdar 109 but lower than those reported by Worrilow et al 57 and by Parmegiani et al, 102 and was significantly lower in the PICSI arm than the ICSI arm (RR 0.963; p < 0.001).…”

“…Our findings on the relationships between HBS and sperm concentration and motility agreed with those of Mokánszki et al 108 and Worrilow et al 57 and we can reasonably assume, therefore, that, despite the limited sample size with data for all assays (n = 131), the associations we uncovered between sperm vitality and DNA integrity could be extended to these and other studies that examined their relationship with HBS. 49,70,104,113 In the spirit of the SAPm (see Report Supplementary Material 2), which sought to explore mechanistic outcomes in relation to clinical outcomes, results are discussed in developmental order and context.…”