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.
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.
Authors: Hoi Chang Lee; Margaret Arny; Daniel Grow; Daniel Dumesic; Rafael A Fissore; Teru Jellerette-Nolan Journal: J Assist Reprod Genet Date: 2014-04-23 Impact factor: 3.412
Authors: Guido de Wert; Björn Heindryckx; Guido Pennings; Angus Clarke; Ursula Eichenlaub-Ritter; Carla G van El; Francesca Forzano; Mariëtte Goddijn; Heidi C Howard; Dragica Radojkovic; Emmanuelle Rial-Sebbag; Wybo Dondorp; Basil C Tarlatzis; Martina C Cornel Journal: Hum Reprod Open Date: 2018-01-12