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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.

Abstract：

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.

journal_name

Fertil Steril

journal_title

Fertility and sterility

authors

Kashir J,Konstantinidis M,Jones C,Heindryckx B,De Sutter P,Parrington J,Wells D,Coward K

doi

10.1016/j.fertnstert.2012.05.002

subject

Has Abstract

pub_date

2012-08-01 00:00:00

pages

423-31

issue

eissn

0015-0282

issn

1556-5653

pii

S0015-0282(12)00538-9

journal_volume

pub_type

杂志文章

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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.