The influence of certain aminoacids and vitamins on post-thaw fish sperm motility, viability and DNA fragmentation

Cabrita, Elsa; Ma, Shanshan; Diogo, Patrícia; Martínez‐Páramo, S.; Sarasquete, Carmen; Dinis, María Teresa

doi:10.1016/j.anireprosci.2011.03.003

Cited by 147 publications

(138 citation statements)

References 33 publications

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“…However, this vitamin did not influence the sperm motility rates of P. lineatus (NAVARRO et al, 2014), S. aurata or D. labrax (CABRITA et al, 2011;MARTÍNEZ-PÁRAMO et al, 2012). These results differ from those found for human semen (ASKARI et al, 1994) and boar (PENA et al, 2003), suggesting that this vitamin is more effective for mammals than for fish.…”

Section: Resultscontrasting

confidence: 56%

Influence of vitamins C and E on the quality of cryopreserved semen Prochilodus brevis (Prochilodontidae, Teleostei)

Almeida‐Monteiro

Oliveira-Araújo

Lopes

et al. 2017

SCA

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Seminal cryopreservation allows the long-term conservation of gametes of various species, including endangered species, such as Prochilodus brevis. However, the application of this biotechnology can cause damage to sperm cells, reducing seminal quality. Thus, we have sought substances that minimize the damage caused by this process, such as antioxidants. Thus, this study aimed to evaluate the association between two cryoprotectants and two vitamins, in different concentrations, on the quality of cryopreserved semen of P. brevis. For cryopreservation, the experiment was performed in two stages. In the first stage, the semen of 10 animals was submitted to six different freezing means, coming from the combination of 5% glucose, two cryoprotectants (Dimethyl sulfoxide [DMSO] or Methyl glycol) and two vitamins (C or E to 0.0001 mg) for cryopreservation. In the second stage, semen samples of eight animals were diluted in 5% glucose and the best cryoprotectant found in the first stage, associated with three different concentrations of vitamins C or E (0.01, 0.001, and 0.0001 mg). In both steps, the in natura and post-thawed samples were submitted to kinetic analysis, morphology, and sperm membrane integrity. The cryopreserved semen with DMSO presented significantly higher results (p < 0.05) than that frozen with Methyl glycol, regardless of the vitamin used. The morphologically normal spermatozoa rate was higher (p < 0.05) in the vitamin-containing samples, however, vitamin E reduced sperm motility rates, independent of the cryoprotectant used. As for vitamin concentrations, higher motility rates were obtained when cryopreserved semen with 0.01 and 0.0001 mg of any of the vitamins. However, the higher concentration had a deleterious effect on the spermatic morphology of P. brevis. Therefore, the glucose associated with DMSO and the lower concentration of vitamin C provides good quality for the post-thawed semen of P. brevis. Key words: Antioxidants. Cryopreservation of semen. ResumoA criopreservação seminal permite a conservação em longo prazo dos gametas de diversas espécies, inclusive as ameaçadas, como a Prochilodus brevis. Contudo, a aplicação dessa biotecnologia pode causar danos às células espermáticas, reduzindo a qualidade seminal. Assim, tem-se buscado substâncias que minimizem os danos causados por esse processo, como os antioxidantes. Deste modo, este estudo objetivou avaliar a associação entre dois crioprotetores e duas vitaminas, em diferentes concentrações, sobre a qualidade do sêmen criopreservado de P. brevis. Para a criopreservação, o experimento foi realizado em duas etapas. Na primeira, o sêmen de 10 animais foi submetido a seis diferentes meios de congelação, oriundos da combinação de glicose 5%, dois crioprotetores (Dimetilsulfóxido [DMSO] ou Metilglicol) e duas vitaminas (C ou E a 0,0001 mg), para a criopreservação. Na segunda etapa, amostras de sêmen de oito animais foram diluídas em Glicose 5% e DMSO (melhor crioprotetor encontrado na primeira etapa), associados a três diferentes conce...

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Section: Resultscontrasting

confidence: 56%

Influence of vitamins C and E on the quality of cryopreserved semen Prochilodus brevis (Prochilodontidae, Teleostei)

Almeida‐Monteiro

Oliveira-Araújo

Lopes

et al. 2017

SCA

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“…These findings show clearly the beneficial effects of our procedure, since several studies in human reported a significant increase of DNA fragmentation in follicles after ovarian cortex cryopreservation with Gosden's procedure [12,33,48]. Supplementation of the collection medium as well as the freezing medium with taurine and L-glutamine, which have been found to play an antioxidant role by reducing cryopreservationinduced oxidative stress, might explain this result [4,18,40]. Although we observed a higher percentage of stroma cells with DNA fragmentation after thawing, this effect remains slight compared with a previous report [49] since it concerns only 16.6 % of stroma cells.…”

Section: Discussionsupporting

confidence: 57%

“…Previous studies reported the beneficial effects of 1,2-propanediol (PrOH) in ovarian tissue cryopreservation [11,14] and raffinose for cold storage of various human tissues before graft [29,51]. Moreover, antioxidant supplements such as taurine and L-glutamine in freezing media have been found to reduce cryoinjuries [4,40]. In an earlier study, follicular growth was observed after subcutaneous xenografting of human ovarian tissue cryopreserved according an original protocol using a serumfree freezing medium supplemented with PrOH and raffinose as well as antioxidants, and a faster freezing program than the usual one [15,39].…”

Section: Introductionmentioning

confidence: 99%

Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Sanfilippo

Canis

Romero

et al. 2012

J Assist Reprod Genet

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Purpose To evaluate the efficiency of an original slow freezing protocol on the quality and function of human ovarian cortex. Methods Human ovarian tissues were cryopreserved using a freezing medium supplemented with propanediol and raffinose as cryoprotectants and antioxidants (L-glutamine, taurine). Samples were then frozen using a faster cooling rate than the usual one. Viability and morphology of follicles, DNA fragmentation in follicles and stroma as well as histology of the vascular endothelium were analyzed before and after freezing/thawing. Moreover, a functional analysis was performed based on the evaluation of follicular growth and development in thawed ovarian tissues that were cultured in vitro. Results Our freezing/thawing protocol allows preservation of a high proportion of viable follicles and the preservation of the different follicle developmental stages (p>0.05 versus fresh control). 70.5±5.2 % of follicles retained an intact morphology after cryopreservation (p=0.04). Stroma cells but not follicles exhibited a slight increase of DNA fragmentation after thawing (p<0.05). Microvessel endothelium within thawed tissues appeared to be preserved. Granulosa cells showed signs of proliferation in follicles cultured for 12 days. Secretion of 17β-oestradiol significantly increased during in vitro culture. Conclusions This protocol leads to good preservation of ovarian integrity and functionality post-thawing and thus appears as a suitable technique of ovarian tissue cryopreservation in clinical settings. Further research could be extended to optimize conditions of in vitro culture.

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“…However, DNA fragmentation in Dicentrarchus labrax semen was higher in treatments containing vitamins than amino acids or control, and in S. aurata semen the antioxidants taurine and hypotaurine significantly reduced DNA fragmentation parameters, protecting DNA against strand breaks (Cabrita et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

“…No significant differences were found in the duration of semen motility with the use of different concentrations of antioxidants; this fact might be due to the short semen handling and manipulation steps reducing ROS action time. Cabrita et al, (2011) working with European sea bass and gilthead sea bream extenders containing several antioxidants suggested a species-specific effect that probably depended on the type of antioxidant added and on its concentration.…”

Section: Cryopreservationmentioning

confidence: 99%

Vitamin E and reduced glutathione in Prochilodus lineatus (curimba) semen cryopreservation (Characiformes: Prochilodontidae)

et al. 2012

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This study investigated the addition of antioxidants vitamin E and reduced glutathione on curimba (Prochilodus lineatus) semen cryopreservation and compared sodium bicarbonate solution and distilled water as activators. The experiment was conducted at the environmental station of CEMIG, in Itutinga-MG, Brazil, between December/2009 and January/2010. Semen samples (n = 7) with semen motility above 80% were diluted in cryoprotectant solutions composed of 10% methanol, 15% lactose and containing different concentrations of antioxidants: 50 (VE50), 100 (VE100) and 250 (VE250) µM of vitamin E, and 0.5 (RG0.5), 1.0 (RG1.0) and 1.5 (RG1.5) mM of reduced glutathione. A solution without antioxidants was used as a control. The semen was diluted at a ratio of 1:4 (100 ìL semen:400 μL cryoprotectant solution). The toxicity of the solutions was evaluated by investigating semen motility after 10 min in the solution. The rest of the diluted semen was placed into 0.5 mL straws maintained in nitrogen vapour for 24 hours and packed into a nitrogen liquid cylinder for four days. The samples were thawed in a water bath at 60°C for 8 s and the rate (%) and duration (s) of semen activation with distilled water or sodium bicarbonate was evaluated. In the toxicity test, we found that vitamin E and reduced glutathione were not toxic to curimba semen at any of the tested concentrations (P>0.05). The duration of motility was longer (P<0.05) in semen activated with sodium bicarbonate 1% (163 ± 11 s), which was considered the best activator for semen under these conditions. No significant differences were found between the cryoprotectant solutions used after freezing (P>0.05). Thus, the antioxidants vitamin E and reduced glutathione did not improve the quality of cryopreserved curimba semen, but they did not cause toxic effects to the semen in natura and they did not decrease its quality during cryopreservation.Este estudo avaliou a adição de antioxidantes vitamina E e glutationa reduzida no sêmen criopreservado de curimba (Prochilodus lineatus) e comparou solução de bicarbonato de sódio e água destilada como ativadores. O experimento foi conduzido na estação ambiental da CEMIG, em Itutinga-MG, entre Dezembro/2009 e Janeiro/2010. Sêmen de sete animais, com motilidade espermática acima de 80%, foi diluído em soluções crioprotetoras compostas por metanol 10% e lactose 15% em diferentes concentrações de antioxidantes: 50 (VE50), 100 (VE100) e 250 (VE250) µM de vitamina E, 0,5 (RG5.5), 1,0 (RG1.0) e 1,5 (RG1.5) mM glutationa reduzida e uma solução controle sem antioxidante. O sêmen foi diluído na proporção de 1:4 (100 µL de sêmen: 400 µL de solução crioprotetora). A toxicidade das soluções foi avaliada pela motilidade espermática após de 10 minutos em solução. O restante do sêmen diluído foi armazenado em palhetas de 0,5 mL mantidos em vapor de nitrogênio por 24 horas e estocado em cilindro de nitrogênio líquido por quatro dias. As amostras foram descongeladas em banho-maria a 60°C por 8 segundos e avaliada a taxa (%) e duração (s) pela ativaçã...

show abstract

The influence of certain aminoacids and vitamins on post-thaw fish sperm motility, viability and DNA fragmentation

Cited by 147 publications

References 33 publications

Influence of vitamins C and E on the quality of cryopreserved semen Prochilodus brevis (Prochilodontidae, Teleostei)

Influence of vitamins C and E on the quality of cryopreserved semen Prochilodus brevis (Prochilodontidae, Teleostei)

Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Vitamin E and reduced glutathione in Prochilodus lineatus (curimba) semen cryopreservation (Characiformes: Prochilodontidae)

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