Store-operated calcium entry in human spermatozoa

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http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/19117 http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/19117 http://eprints.org/relation/isIndexCodesVersionOf http://etheses.bham.ac.uk/id/document/19117 archive 4031 disk0/00/00/32/60 2012-04-19 15:58:36 2019-01-23 09:55:53 2012-04-19 15:58:36 thesis show 0 Nash Katherine Louise Publicover Stephen J. Lefievre Linda University of Birmingham Store-operated calcium entry in human spermatozoa unpub QD QH301 RG RM 10col_life The contribution of store-operated calcium entry (SOCE) in the response of human sperm to progesterone, a steroid secreted from the cumulus cells surrounding the oocyte, has not yet been elucidated. The aim of this study was to investigate the presence of SOCE proteins in human sperm and examine the effects of pharmacological modulation of SOCE on the progesterone-induced biphasic intracellular calcium concentration ([Ca\(^{2+}\)]i) response. STIM (stromal interacting molecule) and Orai, proteins of the SOCE system were detected in human sperm in a similar location to intracellular Ca\(^{2+}\) stores. 2-aminoethyldiphenyl borate (2-APB; SOCE modulator) altered SOCE in human sperm in a bimodal manner as seen in other cell types. Furthermore, 5\(\mu\)M 2-APB potentiated the initial progesterone-induced [Ca\(^{2+}\)]i transient within the neck and midpiece, but not in the flagellum. In the sustained phase of the progesterone-induced [Ca\(^{2+}\)]i response both 5\(\mu\)M 2-APB and 10\(\mu\)M loperamide (another modulator of SOCE) potentiated the [Ca\(^{2+}\)]i response. Higher doses of 2-APB (50-200\(\mu\)M) didn’t potentiate the transient [Ca\(^{2+}\)]i and inhibited the sustained response consistent with reported actions on SOCE. Ryanodine receptors were localised to the neck/midpiece region which suggested that they may mobilise intracellular Ca\(^{2+}\) stores in response to progesterone, leading to activation of STIM/Orai and initiating SOCE. 2012-07 2011-09 graduation eng author unchecked This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. public 249 University of Birmingham School of Biosciences d_ph na 2012-07 graduation