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Sperm parameters, protamine deficiency, and apoptosis in total globozoospermia.

Ghasemzadeh J, Talebi AR, Khalili MA, Fesahat F, Halvaei I, Nabi A, Ashourzadeh S - Iran J Reprod Med (2015)

Bottom Line: Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls (p<0.05).The rate of apoptotic spermatozoa (TUNEL positive) were significantly increased in globozoospermic cases with respect to the controls (17.60 ± 10.72 and 5.95 ± 3.02, respectively, p<0.0001).There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men.

View Article: PubMed Central - PubMed

Affiliation: International Campus, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

ABSTRACT

Background: Globozoospermia is a severe form of teratozoospermia (incidence < 0.1%) in infertile men that is characterized by round headed sperm and acrosomeless in semen.

Objective: To compare the semen parameters, protamine deficiency, and apoptosis in ejaculated spermatozoa between globozoospermic and normozoospermic men.

Materials and methods: Thirty six semen samples were divided into two groups including 15 infertile men with total globozoospermic (> 90% round-headed sperm) and 21 healthy donors with normal spermograms as controls. Semen analysis was performed according to World Health Organization criteria (2010). Sperm protamine deficiency was assessed using Chromomycin A3 (CMA3) staining and the rate of apoptotic spermatozoa was evaluated with TUNEL assay.

Results: Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls (p<0.05). The rate of CMA3-reacted spermatozoa (CMA3+) in globozoospermic men was higher than controls (65.93 ± 11.77 vs. 21.24 ± 7.37, respectively, p<0.0001). The rate of apoptotic spermatozoa (TUNEL positive) were significantly increased in globozoospermic cases with respect to the controls (17.60 ± 10.72 and 5.95 ± 3.02, respectively, p<0.0001). There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men.

Conclusion: Globozoospermic samples contain a higher proportion of spermatozoa with abnormal chromatin packaging and DNA fragmentation than normozoospermic samples. Therefore, in addition to absence of acrosome in the spermatozoa of globozoospermic patients, the high percentage of spermatozoa with immature chromatin and apoptotic marker may be considered as the other etiologies of infertility in these patients.

No MeSH data available.


Related in: MedlinePlus

CMA3 staining for evaluation of sperm protamine deficiency. Bright yellow round-headed sperm cells (CMA3+) show protamine deficiency and yellowish green round-headed sperm cells (CMA3-) show normal protamine content (fluorescent microscopy, × 100 eyepiece magnification).
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Figure 2: CMA3 staining for evaluation of sperm protamine deficiency. Bright yellow round-headed sperm cells (CMA3+) show protamine deficiency and yellowish green round-headed sperm cells (CMA3-) show normal protamine content (fluorescent microscopy, × 100 eyepiece magnification).

Mentions: Chromomycin A3 is a fluorochrome specific for guanosine cytosine-rich sequences and is used for evaluation of the degree of protamination of sperm chromatin (19). To do this test, the air-dried smears were fixed by Carnoy’s solution (methanol/glacial acetic acid, 3:1) for 10 minutes at 4◦C. The slides were stained by CMA3 solution (Sigma, St Louis, MO, USA) (0.25 mg/ml in McIlvaine buffer; 7 ml citric acid, 0.1 M + 32.9 ml Na2HPO4 7H2O 0.2 M, pH 7.0 containing 10 mM MgCl2) for 10 minutes at room temperature. After washing, the slides were mounted by DPX, and then at least 200 spermatozoa were counted under florescent microscopy (BX51, Olympus, Tokyo, Japan) with a 460-nm filter and X100 eyepiece magnification. The percentage of spermatozoa with bright yellow heads (CMA3+ with protamine deficiency) and without brightness (CMA3-with normal protamine content) was evaluated in each sample (Figure 2).


Sperm parameters, protamine deficiency, and apoptosis in total globozoospermia.

Ghasemzadeh J, Talebi AR, Khalili MA, Fesahat F, Halvaei I, Nabi A, Ashourzadeh S - Iran J Reprod Med (2015)

CMA3 staining for evaluation of sperm protamine deficiency. Bright yellow round-headed sperm cells (CMA3+) show protamine deficiency and yellowish green round-headed sperm cells (CMA3-) show normal protamine content (fluorescent microscopy, × 100 eyepiece magnification).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4637114&req=5

Figure 2: CMA3 staining for evaluation of sperm protamine deficiency. Bright yellow round-headed sperm cells (CMA3+) show protamine deficiency and yellowish green round-headed sperm cells (CMA3-) show normal protamine content (fluorescent microscopy, × 100 eyepiece magnification).
Mentions: Chromomycin A3 is a fluorochrome specific for guanosine cytosine-rich sequences and is used for evaluation of the degree of protamination of sperm chromatin (19). To do this test, the air-dried smears were fixed by Carnoy’s solution (methanol/glacial acetic acid, 3:1) for 10 minutes at 4◦C. The slides were stained by CMA3 solution (Sigma, St Louis, MO, USA) (0.25 mg/ml in McIlvaine buffer; 7 ml citric acid, 0.1 M + 32.9 ml Na2HPO4 7H2O 0.2 M, pH 7.0 containing 10 mM MgCl2) for 10 minutes at room temperature. After washing, the slides were mounted by DPX, and then at least 200 spermatozoa were counted under florescent microscopy (BX51, Olympus, Tokyo, Japan) with a 460-nm filter and X100 eyepiece magnification. The percentage of spermatozoa with bright yellow heads (CMA3+ with protamine deficiency) and without brightness (CMA3-with normal protamine content) was evaluated in each sample (Figure 2).

Bottom Line: Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls (p<0.05).The rate of apoptotic spermatozoa (TUNEL positive) were significantly increased in globozoospermic cases with respect to the controls (17.60 ± 10.72 and 5.95 ± 3.02, respectively, p<0.0001).There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men.

View Article: PubMed Central - PubMed

Affiliation: International Campus, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

ABSTRACT

Background: Globozoospermia is a severe form of teratozoospermia (incidence < 0.1%) in infertile men that is characterized by round headed sperm and acrosomeless in semen.

Objective: To compare the semen parameters, protamine deficiency, and apoptosis in ejaculated spermatozoa between globozoospermic and normozoospermic men.

Materials and methods: Thirty six semen samples were divided into two groups including 15 infertile men with total globozoospermic (> 90% round-headed sperm) and 21 healthy donors with normal spermograms as controls. Semen analysis was performed according to World Health Organization criteria (2010). Sperm protamine deficiency was assessed using Chromomycin A3 (CMA3) staining and the rate of apoptotic spermatozoa was evaluated with TUNEL assay.

Results: Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls (p<0.05). The rate of CMA3-reacted spermatozoa (CMA3+) in globozoospermic men was higher than controls (65.93 ± 11.77 vs. 21.24 ± 7.37, respectively, p<0.0001). The rate of apoptotic spermatozoa (TUNEL positive) were significantly increased in globozoospermic cases with respect to the controls (17.60 ± 10.72 and 5.95 ± 3.02, respectively, p<0.0001). There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men.

Conclusion: Globozoospermic samples contain a higher proportion of spermatozoa with abnormal chromatin packaging and DNA fragmentation than normozoospermic samples. Therefore, in addition to absence of acrosome in the spermatozoa of globozoospermic patients, the high percentage of spermatozoa with immature chromatin and apoptotic marker may be considered as the other etiologies of infertility in these patients.

No MeSH data available.


Related in: MedlinePlus