Preliminary results on ascorbic acid and lysine suppression of clastogenic effect of deep-frozen sperm of the Russian sturgeon (Acipenser gueldenstaedti)

Mirzoyan, A. V.; Nebesikhina, N.A.; Voynova, N. V.; Chistyakov, V. A.

doi:10.1016/j.ijrefrig.2005.07.008

Cited by 39 publications

(22 citation statements)

References 16 publications

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“…According to Dziewulska et al (2011) the best cryoprotectant for maintaining cytoplasm-membrane integrity, mitochondrial function and motility patterns is DMSO supplemented with antioxidants and cytoplasm-membrane stabilisers, such as BSA. In salmonids, this maintains the lipid composition of the plasma membrane and increases the stability of the spermatozoa (Baynes & Scott, 1987;Lahnsteiner et al, 1996a;Cabrita et al, 2001;Bergeron & Manjunath, 2006;Mirzoyan et al, 2006). Furthermore, the incorporation of sugars like sucrose and glucose into the cryopreservation medium has been used effectively for freezing (Mansour et al, 2006;Tekin et al, 2007) spermatozoa of O. mykiss, asp Leuciscus aspius (L. 1758) and S. salar (Lahnsteiner et al, 1996b;Babiak et al, 1998;Lahnsteiner, 2000;Kusuda et al, 2005;Mansour et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of Atlantic salmonSalmo salarsperm: effects on sperm physiology

Figueroa

Valdebenito

Merino

et al. 2016

Journal of Fish Biology

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The objective of this study was to determine the effect of freezing on the function in Atlantic salmon Salmo salar spermatozoa. The semen was frozen in Cortland's medium + 1.3M dimethyl sulphoxide + 0.3M glucose + 2% bovine serum albumin (final concentration) in a ratio of 1:3 (semen:cryoprotectant) as the treatment (T) and fresh semen as the control (F). Straws of 0·5 ml of sperm suspension were frozen in 4 cm of N2 L. They were thawed in a thermoregulated bath (40° C). After thawing, the percentage of spermatozoa with fragmented DNA [transferase dUTP (deoxyuridine triphosphate) nick-end labelling (TUNEL)], plasma membrane integrity (SYBR-14/PI) and mitochondrial membrane potential (ΔΨMMit, JC-1) were evaluated by flow cytometry and motility was evaluated by optical microscope under stroboscopic light. The fertilization rates of the control and treatment semen were tested at a sperm density of 1·5 × 10(7) spermatozoa oocyte(-1) , by observation of the first cleavages after 16 h incubation at 10° C. In the cryopreserved semen (T), the mean ± s.d. DNA fragmentation was 4·8 ± 2·5%; plasma membrane integrity 75·2 ± 6·3%; mitochondrial membrane potential 51·7 ± 3·6%; motility 58·5 ± 5·3%; curved line velocity (VCL ) 61·2 ± 17·4 µm s(-1) ; average-path velocity (VAP ) 50·1 ± 17·3 µm s(-1) ; straight-line velocity (VSL ) 59·1 ± 18·4 µm s(-1) ; fertilization rate 81·6 ± 1·9%. There were significant differences in the plasma membrane integrity, mitochondrial membrane potential, motility, fertilization rate, VCL , VAP and VSL compared with the controls (P < 0·05). Also the mitochondrial membrane potential correlated with motility, fertilization rate, VCL and VSL (r = 0·75; r = 0·59; r = 0·77 and r = 0·79, respectively; P < 0·05); and the fertilization rate correlated with VCL and VSL (r = 0·59 and r = 0·55, respectively).

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Section: Discussionmentioning

confidence: 99%

Cryopreservation of Atlantic salmonSalmo salarsperm: effects on sperm physiology

Figueroa

Valdebenito

Merino

et al. 2016

Journal of Fish Biology

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“…Future studies should focus on controlling ATP content in cryopreserved semen similarly to that which took place in studies conducted by Williot et al (2000), during which the authors attempted to find sources of energy in Siberian sturgeon semen. The influence of antioxidants on the cryopreservation process of semen should also be analyzed, as confirmed by recent studies, the addition of ascorbic acid to the freezing solution may prevent the accumulation of active forms of oxygen and thus, influence an increase of sperm motility and their ability to fertilize (Mirzoyan et al, 2006).…”

Section: Discussionmentioning

confidence: 88%

“…When describing the decrease in the quality of sperm after cryopreservation, Dzyuba et al (1999) suggested exposing the semen to the effects of oxygen in order to maintain sperm motility. The influence of antioxidants on the cryopreservation process of semen should also be analyzed, as confirmed by recent studies, the addition of ascorbic acid to the freezing solution may prevent the accumulation of active forms of oxygen and thus, influence an increase of sperm motility and their ability to fertilize (Mirzoyan et al, 2006). Future studies should focus on controlling ATP content in cryopreserved semen similarly to that which took place in studies conducted by Williot et al (2000), during which the authors attempted to find sources of energy in Siberian sturgeon semen.…”

Section: Discussionmentioning

confidence: 89%

Cryopreservation of Siberian sturgeon (Acipenser baerii , Brandt, 1869) and sterlet (Acipenser ruthenus , Linnaeus, 1758) semen and its influence on sperm motility parameters assessed using a computer-assisted sperm analysis (CASA) system

Sieczyński¹,

Cejko

Grygoruk³

2015

J. Appl. Ichthyol.

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Summary Using a computer assisted sperm analysis (CASA) system, sperm motility parameters were determined in semen of the Siberian sturgeon (Acipenser baerii) and sterlet (Acipenser ruthenus) prior to and after cryopreservation. Semen was collected from 15 Siberian sturgeon and 15 sterlet males, diluted 1:1 (sperm:freezing medium) in 23.4 mm sucrose, 0.25 mm KCl, 30 mm Tris (pH of 8.0) and 10% methanol, loaded in 0.25 ml straws and frozen over liquid nitrogen vapor. Post‐thaw sperm motility was activated in 10 mm Tris buffer (pH 8.5), containing 20 mm NaCl and 2 mm CaCl2. A significant decrease in the percentage of motile sperm (MOT: from 41.3% to 25.3%), curvilinear velocity (VCL: from 121.9 to 112.2 μm s−1), amplitude of lateral head displacement (ALH: from 10.4 to 5.4 μm) and momentum (MOM: from 4591 to 1038 m s−1) was observed as effect of the cryopreservation process in the Siberian sturgeon semen. A significant decrease in the values of most CASA parameters, including MOT, VAP, VCL, VSL, ALH and MOM, were also noted following the cryopreservation of sterlet semen. In the sterlet, an additional decrease in VAP (from 102.4 to 80.3 μm s−1) and VSL (from 86.8 to 69.8 μm s−1) values was noted after cryopreservation when compared to fresh semen. Unfortunately, the cryopreservation of semen from both species led to a drop in CASA parameters, which are important to the successful fertilization of eggs. MOT and VCL parameters were the most affected in these species. They can be used as sensitive criteria to check changes in motility.

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“…Ascorbic acid, also known as Vitamin C, has also been associated with its antioxidative action in protecting spermatozoa plasma membrane integrity. Mirzoyan, Nebesikhina, Voynova and Chistyakov () showed that addition of ascorbic acid (0.1 M) to the cryoprotective medium containing 15% DMSO increased motility and fertilizing ability of the frozen/thawed sperm of Russian sturgeon ( Acipenser gueldenstaedti ). In this study, the tested ascorbic acid (vitamin C) concentrations had no effect on post‐thaw motility of sperm.…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants, antioxidant, membrane stabilizer, equilibration time and dilution ratio on sperm motility and fertility

2015

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Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL À1 ) and ascorbic acid (0, 2.5, 5 and 10 U mL À1 ), on the post-thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL À1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post-thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post-thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post-thaw motility (P < 0.05). The motility of frozen-thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL À1 BSA significantly improved post-thaw motility (P < 0.05), but ascorbic acid did not improve post-thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 AE 5.75) and hatching rates (28.2 AE 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL À1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.

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Preliminary results on ascorbic acid and lysine suppression of clastogenic effect of deep-frozen sperm of the Russian sturgeon (Acipenser gueldenstaedti)

Cited by 39 publications

References 16 publications

Cryopreservation of Atlantic salmonSalmo salarsperm: effects on sperm physiology

Cryopreservation of Atlantic salmonSalmo salarsperm: effects on sperm physiology

Cryopreservation of Siberian sturgeon (Acipenser baerii , Brandt, 1869) and sterlet (Acipenser ruthenus , Linnaeus, 1758) semen and its influence on sperm motility parameters assessed using a computer-assisted sperm analysis (CASA) system

Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants, antioxidant, membrane stabilizer, equilibration time and dilution ratio on sperm motility and fertility

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