Demonstration of the effect of epidermal growth factor on ram sperm parameters using two fluorescent assays

ABSTRACT: The goal of this study was to examine the effect of epidermal growth factor (EGF) on sperm viability using two fluorescent techniques and to analyze the obtained results in relation to sperm motility, determined by subjective estimation. Fresh ram semen diluted in a Biladyl commercial extender was cooling stored (at 4 °C in a fridge) for four days in the presence of EGF at doses of 0, 10, 100, 200 or 400 ng/ml. Thereafter, sperm samples were analyzed for progressive motility (Motility test) and membrane integrity using two fluorescent techniques: SYBR-14/PI (Method 1) or PI/DAPI (Method 2). Application of Method 1 did not detect an effect of EGF at any concentration on sperm membrane integrity. A positive effect of EGF (200 ng/ml) on sperm membrane integrity was found using Method 2 of staining, and this result was confirmed by the sperm motility test, which demonstrated an EGF-stimulating effect (200 or 400 ng/ml) on a percentage of progressively moving spermatozoa. Strong positive correlations between Methods 1 and 2 (r = 0.785), Method 1 and Motility (r = 0.803), Method 2 and Motility (r = 0.699), as well as between both techniques taken together and the Motility test (r = 0.853) were found. Regression analysis confirmed that Method 2 was more exact than Method 1, and the results obtained with Method 2 are comparable with those of the Motility test. Dependence of the viability or motility on EGF concentrations (linear regression function) was significant only for Method 2 or the Motility test. The obtained results suggest a stimulating effect of EGF (at higher concentrations) on ram sperm functions (viability/membrane integrity and motility). Furthermore, they indicate substantial differences between two fluorescent techniques in the determination of sperm membrane integrity. Only the data obtained using PI/DAPI were confirmed by a functional Motility test. These findings suggest that the technique chosen for analysis of sperm viability can influence the conclusion concerning the effects of the treatment on sperm function.

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Ram Sperm Motility Parameters under The Influence of Epidermal Growth Factor

Veterinary Medicine International ◽

10.4061/2011/642931 ◽

2011 ◽

Vol 2011 ◽

pp. 1-5 ◽

Cited By ~ 2

Author(s):

Eliška Špaleková ◽

Alexander V. Makarevich ◽

Norbert Lukáč

Keyword(s):

Growth Factor ◽

Epidermal Growth Factor ◽

Sperm Motility ◽

Computer Assisted ◽

Female Reproduction ◽

Male And Female ◽

Progressive Movement ◽

Epidermal Growth ◽

Ram Sperm ◽

Motility Parameters

Epidermal growth factor (EGF) is one of the important cytokines that play a role in fertility. It is known that EGF affects both male and female reproduction, but its effect on sperm parameters is not fully understood. Up to the present, the effect of EGF on ram sperm motility parameters has not been published. We analyzed motility parameters of ejaculates after 24, 48, and 72 hours from the EGF addition. EGF was added to chilled ram sperm at concentrations of 0, 100, 200, and 400 ng⋅ml−1. Analyses were realized using computer, assisted semen analyzer (CASA)—Hamilton Thorn motility analyzer (version 7). The effect of EGF was already visible after 30 min of incubation. Significant effect on ram sperm total motility and progressive movement was observed at higher EGF concentrations after 48 h of incubation. Our results show that EGF affects sperm motility parameters depending on concentration and time of exposure.

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Beneficial Role of Ginger Powder (Zingiber officinale) against Acephate-induced Reprotoxicity in Adult Male Rats

European Journal of Medicinal Plants ◽

10.9734/ejmp/2021/v32i830410 ◽

2021 ◽

pp. 50-59

Author(s):

V. Madhavi ◽

U. Kanchana Ganga ◽

S. B. Sainath ◽

B. Kishori

Keyword(s):

Body Weight ◽

Sperm Motility ◽

Sperm Count ◽

Membrane Integrity ◽

Male Rats ◽

Sperm Viability ◽

Marker Enzymes ◽

Group I ◽

Sperm Membrane

Aims: The present study was aimed to investigate the protective role of ginger against acephate-induced testicular toxicity in adult rats. Methodology: Rats were allocated into four groups where animals in group I served as controls, while animals in group II, III and group IV were treated as experimental rats. Rats in groups II, III and IV were treated with acephate (50mg/kg body weight), ginger (100mg/kg body weight) and combination of both acephate and ginger, respectively over a period of 60 days. After completion of experimental period sperm count, sperm viability, sperm motility, sperm membrane integrity, testicular steroidogenic marker enzymes (3β-HSD and 17β-HSD, serum testosterone and testicular architecture was performed in both control and experimental rats. Results: Relative weights of reproductive organs, sperm count, sperm viability, sperm motility and sperm membrane integrity were significantly decreased in acephate treated rats over controls. Acephate administration also reduced the circulatory levels of testosterone associated with a significant reduction in the testicular steroidogenic marker enzymes (3β-HSD and 17β-HSD) in rats. The testicular architecture was disrupted in acephate intoxicated rats. In contrast, ginger administration significantly recovered the acephate-induced suppressed selected reproductive parameters with increased circulatory levels of testosterone and restoration of sperm endpoints in as compared to acephate alone treated rats. No significant changes were observed in any of the selected reproductive endpoints in ginger treated rats as compared to controls. Conclusion: The results can be concluded that supplementation of ginger mitigates the negative effects of acephate on male reproductive health via amelioration of testicular setroidogenesis and spermatogenesis and epididymal sperm maturation events in rats.

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Protease activated receptor 2 and epidermal growth factor receptor are involved in the regulation of human sperm motility

Asian Journal of Andrology ◽

10.1111/j.1745-7262.2007.00289.x ◽

2007 ◽

Vol 9 (5) ◽

pp. 690-696 ◽

Cited By ~ 8

Author(s):

Karina Zitta ◽

Martin Albrecht ◽

Stephan Weidinger ◽

Artur Mayerhofer ◽

Frank Köhn

Keyword(s):

Epidermal Growth Factor Receptor ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Sperm Motility ◽

Growth Factor Receptor ◽

Human Sperm ◽

Epidermal Growth ◽

Protease Activated Receptor 2

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Inactivation of a sperm motility gene by insertion of an epidermal growth factor receptor transgene whose product is overexpressed and compartmentalized during spermatogenesis.

Genes & Development ◽

10.1101/gad.5.8.1395 ◽

1991 ◽

Vol 5 (8) ◽

pp. 1395-1406 ◽

Cited By ~ 31

Author(s):

G T Merlino ◽

C Stahle ◽

C Jhappan ◽

R Linton ◽

K A Mahon ◽

...

Keyword(s):

Epidermal Growth Factor Receptor ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Sperm Motility ◽

Growth Factor Receptor ◽

Epidermal Growth

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Effect of different concentrations of trehalose and glycerol on the freezability of ram semen using soybean lecithin-based diluents

Animal Production Science ◽

10.1071/an13431 ◽

2015 ◽

Vol 55 (5) ◽

pp. 666 ◽

Cited By ~ 1

Author(s):

Zh. Bohlool ◽

M. Mohammadi ◽

M. Roostaei-Ali Mehr ◽

N. Ghavi Hossein-Zadeh

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Soybean Lecithin ◽

Membrane Integrity ◽

Sperm Viability ◽

Plasma Membrane Integrity ◽

Main Effect ◽

Acrosome Integrity ◽

Ram Sperm ◽

Desirable Effect

This study was conducted to determine the effect of different levels of trehalose and glycerol on ram sperm cryosurvival using lecithin-based diluents. Ejaculates were collected from four rams, pooled after initial evaluation, diluted with Tris-soybean lecithin extender and split into nine equal parts. A total of 0 (T0), 50 (T50) or 100 (T100) mM of trehalose and 3% (G3), 5% (G5) or 7% (G7) of glycerol were added to each part. Sperm motility, viability, plasma membrane integrity and acrosome integrity were evaluated immediately after thawing (0 h), and subsequently after 3 h and 6 h post-thawing incubation at 37°C. Results indicated that there was interaction between trehalose and glycerol on sperm motility. In addition, interaction of trehalose and glycerol with incubation time on sperm motility, viability, plasma membrane integrity and acrosome integrity was not significant (P > 0.05). Sperm motility was greatest in the sperm treated with 100 mM trehalose and 7% glycerol (27%; P < 0.05). The effect of trehalose was significant on viability and plasma membrane integrity of ram spermatozoa (P < 0.05). The main effect of trehalose showed that sperm viability was higher in T100 (47.06%) than T50 (53.96%; P < 0.05). The highest membrane integrity was observed in T100 (47.04%; P < 0.05). Membrane integrity was higher (P < 0.05) in G5 (49.97%) than G3 (41.56%) and there was no difference between G7 (46.86%) and G3 (41.56%; P > 0.05). The best sperm viability and plasma membrane integrity was observed at 0 h (65.75% and 51.58%, respectively). It was concluded that simultaneous use of 7% glycerol and 100 mM trehalose had a desirable effect on motility of ram frozen–thawed sperm.

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