Relationship between phospholipase C-zeta, semen parameters, and chromatin status

Figures & data

Table 1. Comparison of semen parameters between men with normal (n=32) and abnormal (n=23) semen parameters.

Parameters	Normal semen parameters	Abnormal semen parameters	P-value
Sperm concentration (10^6/ml)	74.84±5.21	26.82±4.62	0.001
Total sperm count (10^6/ejaculate)	350.72±30.78	104.54±22.52	0.001
Total sperm motility (%)	62.65±2.08	35.56±3.46	0.001
Progressive sperm motility (%)	39.77±1.98	14.41±2.42	0.001
Abnormal sperm morphology (%)	95.43±0.21	98.26±0.20	0.001
Semen volume (ml)	5.12±0.38	3.63±0.31	0.004

Data are presented as mean ± SEM and analyzed by independent-samples t-Test.

Figure 1. Comparison of mean percentage of sperm PLCζ, SDF, and protamine deficiency between men with normal and abnormal semen parameters. The mean percentage of PLCζ positive spermatozoa was significantly lower, and percentage of SDF and protamine deficient spermatozoa were significantly higher in men with abnormal semen parameters compared to men with normal semen parameters, respectively. SDF: sperm DNA fragmentation; PLCζ: phospholipase C-zeta. # shows significant difference between the two groups at p<0.05.

Figure 2. Flow cytometry analysis of percentages of PLCζ positive spermatozoa in two semen samples with normal (A) and abnormal (B) semen parameters. Percentage of PLCζ positive spermatozoa was higher in individuals with normal parameters compared to abnormal parameters. Fluorescence images of spermatozoa stained with TUNEL (C) and CMA3 staining (D) for DNA fragmentation and protamine deficiency, respectively. Spermatozoa with green staining (TUNEL⁺) and bright yellow staining (CMA³⁺) were considered as protamine deficient and DNA fragmented, respectively. PLCζ: phospholipase C-zeta; CMA3: chromomycin A3; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labelling.

Table 2. Correlation between percentage of spermatozoa presenting DNA fragmentation, protamine deficiency, and PLCζ with sperm concentration, sperm motility, and sperm abnormal morphology (n=55).

Parameters	Concentration (%)		Motility (%)	Abnormal morphology (%)
	R	0.476**	0.430**	-0.317*
PLCζ positive spermatozoa (%)	P	0.001	0.001	0.012
	R	-0.173	-0.629**	0.434**
DNA fragmented spermatozoa (%)	P	0.207	0.001	0.001
	R	-0.343*	-0.342*	0.475**
Protamine deficient spermatozoa (%)	P	0.011	0.011	0.001

PLCζ: phospholipase C-zeta. **Correlation is significant at the 0.01 level (2-tailed); *Correlation is significant at the 0.05 level (2-tailed).

Table 3. Multiple regression analysis between sperm parameters and percentage of PLCζ positive spermatozoa.

	Adjusted OR (95% CI)	P-value
Sperm concentration	1 (0.97-1.08)	0.33
Sperm motility (%)	1 (0.95-1.11)	0.461
Abnormal sperm morphology (%)	0.25 (0.08-0.78)	0.018
PLCζ positive spermatozoa (%)
%≤50	0.4 (0.04-4.8)	0.49
%>51^ref	-	-

PLCζ: phospholipase C-zeta. Dependent variable in multinomial logistic regression with two categories; abnormal parameters were considered as reference category. OR: odd ratio; 95% CI: 95% confidence interval.

Figure 3. Correlation between percentage of PLCζ with DNA fragmentation (r = -0.3; p=0.026) and protamine deficiency (r= -0.05; p=0.702) in total population (32 men with normal semen parameters and 23 men with abnormal semen parameters). PLCζ: phospholipase C-zeta.