Critically Evaluate the Evidence for PLC ZETA: Essay Writing

Internal Code: MAS 5960

SPERM QUALITY PARAMETERS AND SPERMATOGENESIS NOT AFFECTED BY PLC?

Epididymal sperm from Plcz1 –/– males (Plcz1 – sperm) had normal motility, normal viability and hyperactivity, which reflect their ability to undergo capacitation (fig.B,C,D) . Furthermore, analysis of Plcz1 –/– male testes determined no deficiency in spermatogenesis (fig. A).In response to progesterone or ionomycin, the ability of Plcz1 – sperm to undergo acrosome reaction was uneventful from the sperm of wild type animals (fig H). Therefore loss of PLC? has no likely effects on parameters connected with the ability of the sperm to bind/fuse with the egg and also is likely to have no effects on spermatogenesis. Characterization of sperm from homozygote Plcz1 em1Jparr and Plcz1 em2Jparr mice.(A)Stain hematoxylin/Eosin used for analysis of testis sections. (B) Flow cytometry used to plot density of sperm stained with SYBR14 (LIVE SPERM) and propidium iodide .Around 10,000 events collected per group. (C) Olympus BX40 Microscope was used to obtain stained sperm. Quantification of data from B. (D-G) with use of computer assisted sperm analysis (CASA), Analysis of sperm motility is done. (H) Treatment with progesterone or ionomycin , An acrosome reaction was initiated and by lack of reactivity towards fluorescently labelled lectin PNA, reacted sperm were further determined. Relatively images of unreacted and reacted sperm were shown. LOSS OF PLC? INVALIDATES THE SPERM TO TRIGGER Ca 2+ OSCILLATIONS IN EGG. We examine whether Plcz1 – sperm can trigger Ca 2+ oscillations in wild-type eggs, using intracytoplasmic sperm injection (ICSI).Injection of Plcz1 + sperm induced Ca 2+ oscillations pointing to those seen at fertilization in almost all the eggs (Fig. A,B) (Wakai et al., 2011). Later it was noted that injected sperm from Plcz1 –/– line did not induce Ca 2+ oscillations. Although it were triggered by injection of Plcz1 WT cRNA (Fig.C). By use of zona pellucida free eggs in vitro fertilization (IVF) , It was confirmed that there was an inability of Plcz1 – sperm to trigger Ca 2+ oscillations. (Fig.D,E).during the process 16 out of 34 eggs displayed Ca 2+ oscillations in IVF with Plcz1 + sperm. Out of 40 eggs which were subjected to IVF with Plcz1 em1Jparr -derived sperm. Not a single egg displayed such oscillations, with only one egg showing single Ca 2+ transient (Fig.D,E). This in return could be the first evidence stating that PLC? can be the sole physiological trigger of the Ca 2+ oscillations, which could be responsible for egg activation. (Wakai et al., 2011)