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摘要
Does motile sperm organelle morphology examination (MSOME) affect levels and localization patterns of the oocyte activation factor phospholipase C zeta (PLC?) in globozoospermic sperm with and without an acrosomal bud? %26lt;br%26gt;MSOME identified round-headed globozoospermic sperm with increased levels of PLC? relative to sperm from the same sample that did not undergo MSOME, and identified novel patterns of PLC? localization in sperm exhibiting an acrosomal bud. %26lt;br%26gt;Absence or reduction in the level of PLC? in the sperm head, abnormal localization patterning, or defective functional ability as a result of PLC? gene mutation, have been linked to certain types of human male factor infertility in which oocyte activation is deficient. It has been determined that a subpopulation of sperm (1) from a patient exhibiting 100 globozoospermia presented with an acrosome bud upon MSOME. A cycle of intracytoplasmic morphologically selected sperm injection, carried out with sperm exhibiting an acrosomal bud led to pregnancy and birth of a healthy baby boy, without the use of assisted oocyte activation (AOA). %26lt;br%26gt;Immunofluorescent analysis of PLC? in globozoospermic sperm from three patients, before and after MSOME. %26lt;br%26gt;Quantitative immunofluorescence was used to investigate PLC? levels and localization patterns in individual sperm (n 1 patient) identified by MSOME and isolated by micromanipulation, and presenting with and without the acrosomal bud. A secondary aim was to investigate levels and localization patterns of PLC? in sperm before and after MSOME from two other globozoospermic men. %26lt;br%26gt;Non-globozoospermic control sperm exhibited characteristic localization patterns of PLC? immunofluorescence. Completely round-headed globozoospermic sperm from patients 13 were either devoid of PLC? immunofluorescence, or exhibited an abnormal, punctate, pattern of PLC? localization. PLC? immunofluorescence in sperm exhibiting an acrosomal bud was observed in the midpiece with varying fluorescent intensity and was detected in 28.5 of such sperm. The majority of sperm with an acrosomal bud (43.0) exhibited punctate patterns of PLC? localization within the sperm head. A further 28.5 of sperm exhibited PLC? in both the head and the midpiece. Total levels of PLC?, and the proportions of sperm exhibiting PLC? immunoreactivity, showed significant variance (P 0.05) amongst control [45.8 arbitrary units (a.u.) and 95.7, respectively], non-MSOME-selected (25.9 a.u. and 46.1, respectively) and MSOME-selected globozoospermic sperm (33.4 a.u. and 65.0, respectively). Total levels of PLC? immunofluorescence, and proportions of sperm exhibiting PLC? immunoreactivity, in control sperm was significantly higher (P 0.05) compared with non-MSOME-selected sperm, but not significantly different from MSOME-selected sperm. %26lt;br%26gt;The low numbers of sperm analysed may not be ideal for conclusive statistical analysis. Evaluation of the effects of MSOME on morphologically normal sperm would confirm conclusions. %26lt;br%26gt;The present findings provide hope for the future treatment of globozoospermia without the need for AOA, and provide further evidence for the clinical application of PLC? as a therapeutic and prognostic tool. %26lt;br%26gt;The research described herein was funded by the Nuffield Department of Obstetrics and Gynaecology, University of Oxford. The authors report no conflict of interest.
- 出版日期2012-11
