Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells
Objective: To examine the underlying factors leading to infertility in a male patient from whom phospholipase C zeta H398P (PLCζ H398P, histidine > proline) and PLCζ H233L (histidine > leucine) mutations were previously identified. Desi...
| Main Authors: | , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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2012
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| _version_ | 1797065659830501376 |
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| author | Kashir, J Konstantinidis, M Jones, C Heindryckx, B De Sutter, P Parrington, J Wells, D Coward, K |
| author_facet | Kashir, J Konstantinidis, M Jones, C Heindryckx, B De Sutter, P Parrington, J Wells, D Coward, K |
| author_sort | Kashir, J |
| collection | OXFORD |
| description | Objective: To examine the underlying factors leading to infertility in a male patient from whom phospholipase C zeta H398P (PLCζ H398P, histidine > proline) and PLCζ H233L (histidine > leucine) mutations were previously identified. Design: Laboratory-based study. Setting: University laboratory. Patient(s): An infertile 38-year-old man with significantly impaired oocyte activation ability. Intervention(s): Minisequencing of individual sperm for PLCζ H398P and PLCζ H233L, and investigation of localization patterns arising from the expression of fluorescently tagged PLCζ isoforms in HEK293T cells. Main Outcome Measure(s): The presence/absence of PLCζ H398P and PLCζ H233L determined in individual sperm (n = 12 sperm), and localization of fluorescent mutant PLCζ isoforms quantified in HEK293T cells. Result(s): Sperm possessed either PLCζ H233L or PLCζ H398P, but never both at the same time. Fluorescent PLCζ H233L and PLCζ H233L+H398P (both mutations together) localized to discrete regions in HEK293T cytoplasm but not the plasma membrane. Fluorescence statistically significantly varied between constructs such that PLCζ WT > mutant isoforms at both 48- and 56-hour time points. Fluorescent-PLCζ H233L+H398P exhibited a statistically significantly reduced level of fluorescence compared with PLCζ H398P at 48 hours but not 56 hours. Conclusion(s): Both H398P and H233L mutations are present on different alleles and do not alter PLCζ localization in HEK293T cells. Loss-of-activity mutations in PLCζ may contribute not only toward male infertility but also male subfertility in cases where PLCζ is mutated on a single allele. Copyright © 2012 American Society for Reproductive Medicine, Published by Elsevier Inc. |
| first_indexed | 2024-03-06T21:31:44Z |
| format | Journal article |
| id | oxford-uuid:44e9a243-c81c-4930-91c0-87142cf5abd2 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:31:44Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | oxford-uuid:44e9a243-c81c-4930-91c0-87142cf5abd22022-03-26T15:04:32ZCharacterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cellsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:44e9a243-c81c-4930-91c0-87142cf5abd2EnglishSymplectic Elements at Oxford2012Kashir, JKonstantinidis, MJones, CHeindryckx, BDe Sutter, PParrington, JWells, DCoward, KObjective: To examine the underlying factors leading to infertility in a male patient from whom phospholipase C zeta H398P (PLCζ H398P, histidine > proline) and PLCζ H233L (histidine > leucine) mutations were previously identified. Design: Laboratory-based study. Setting: University laboratory. Patient(s): An infertile 38-year-old man with significantly impaired oocyte activation ability. Intervention(s): Minisequencing of individual sperm for PLCζ H398P and PLCζ H233L, and investigation of localization patterns arising from the expression of fluorescently tagged PLCζ isoforms in HEK293T cells. Main Outcome Measure(s): The presence/absence of PLCζ H398P and PLCζ H233L determined in individual sperm (n = 12 sperm), and localization of fluorescent mutant PLCζ isoforms quantified in HEK293T cells. Result(s): Sperm possessed either PLCζ H233L or PLCζ H398P, but never both at the same time. Fluorescent PLCζ H233L and PLCζ H233L+H398P (both mutations together) localized to discrete regions in HEK293T cytoplasm but not the plasma membrane. Fluorescence statistically significantly varied between constructs such that PLCζ WT > mutant isoforms at both 48- and 56-hour time points. Fluorescent-PLCζ H233L+H398P exhibited a statistically significantly reduced level of fluorescence compared with PLCζ H398P at 48 hours but not 56 hours. Conclusion(s): Both H398P and H233L mutations are present on different alleles and do not alter PLCζ localization in HEK293T cells. Loss-of-activity mutations in PLCζ may contribute not only toward male infertility but also male subfertility in cases where PLCζ is mutated on a single allele. Copyright © 2012 American Society for Reproductive Medicine, Published by Elsevier Inc. |
| spellingShingle | Kashir, J Konstantinidis, M Jones, C Heindryckx, B De Sutter, P Parrington, J Wells, D Coward, K Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title | Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title_full | Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title_fullStr | Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title_full_unstemmed | Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title_short | Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
| title_sort | characterization of two heterozygous mutations of the oocyte activation factor phospholipase c zeta plcζ from an infertile man by use of minisequencing of individual sperm and expression in somatic cells |
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