Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch (Perca flavescens)

Jenkins, Jill A.; Draugelis-Dale, Rassa O.; Pinkney, Alfred E.; Iwanowicz, Luke R.; Blazer, Vicki S.

doi:10.1016/j.theriogenology.2014.11.028

Cited by 6 publications

(2 citation statements)

References 57 publications

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“…Spermatozoa DNA fragmentation was determined with acridine orange and expressed by the median DNA fragmentation index (Myromslien et al., 2019). Orange fluorescence marked denatured DNA, whereas intact DNA was stained with green fluorescence (Jenkins et al., 2015).…”

Section: Methodsmentioning

confidence: 99%

Nanocarried antioxidants in freezing extenders for boar spermatozoa

et al. 2021

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In swine, most artificial inseminations (AI) are routinely conducted with cooled spermatozoa (Waberski et al., 2019), since AI with frozen spermatozoa yields inferior performance (Yeste, 2016).That may result from injuries in spermatozoa attributed to cold shock during freezing and thawing. Also, boar spermatozoa have high content of unsaturated fatty acids in their plasma membrane and limited intracellular antioxidant mechanisms, being prone to oxidative stress due to lipoperoxidation and to the production of reactive oxygen species (ROS) (Cerolini et al., 2000). Thus, inclusion of antioxidants in extenders for sperm freezing can mitigate oxidative stress.One of such antioxidants would be vitamin E, which is a natural component of spermatozoa's antioxidant system (Lenzi et al., 1996).Positive effects of vitamin E on post-thawing quality have been reported for spermatozoa of men (Taylor et al., 2009), bulls (Nasiri et al., 2012) and boars (Peña et al., 2003;Satorre et al. 2007). Other potentially efficient antioxidant is resveratrol, a molecule found in grapes which has anti-inflammatory properties, but low stability and solubility (Vitaglione et al., 2005). Positive effects of resveratrol were described for human (Branco et al., 2010), bull (Bucak et al., 2015 and buffalo spermatozoa (Longobardi et al., 2017), but negative effects were reported for ram spermatozoa (Silva et al., 2012). Data for boar spermatozoa revealed no effect on post-thawing quality, but positive effects on in vitro oocyte penetration (Gadani et al., 2017).

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Section: Methodsmentioning

confidence: 99%

Nanocarried antioxidants in freezing extenders for boar spermatozoa

et al. 2021

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“…A 30‐s incubation of 10 μl thawed spermatozoa diluted in 5 μl of TNE (0.01 M Tris‐HCl, 0.15 M NaCl, 0.001 M disodium EDTA, pH 7.2) with addition of 10 μl of 1% (v/v) Triton X‐100 was performed, immediately following which, acridine orange dye was added and flow cytometer reading taken within 2 min. Spermatozoa were classified as intact (green) or fragmented (orange/red; Jenkins, Draugelis‐Dale, Pinkney, Iwanowicz, & Blazer, ) with the rate of DNA fragmentation calculated as: [Number of spermatozoa with fragmented DNA/(Number of spermatozoa with intact DNA + Number of spermatozoa with fragmented DNA)] × 100.…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation of sperm in annual fish Austrolebias minuano (Cyprinodontiformes; Rivulidae)

et al. 2019

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The effects of the cryoprotectants dimethyl sulfoxide, glycerol and methyl glycol at different concentrations on Austrolebias minuano sperm quality parameters were evaluated in this study. The cellular kinetic parameters, determined using flow cytometry, indicated the best results with the samples cryopreserved with 7.5% methyl glycol. Dimethyl sulfoxide concentrations of 7.5% and glycerol concentrations of 10%, 12.5% and 15% demonstrated the least benefit across all evaluated sperm kinetic parameters. When assessed using flow cytometry, a concentration of 7.5% methyl glycol similarly showed better sperm kinetic parameters than 12.5% dimethyl sulfoxide. We conclude that cryoprotectants, especially methyl glycol, are effective for the preservation of sperm quality in A. minuano. However, high sensitivity of spermatozoa against glycerol was observed in these studies; thus, it is not recommended for cryopreservation purposes. K E Y W O R D Scryoprotectants, extinction, killifish, preservation

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Reproductive parameters of double transgenic zebrafish (Danio rerio) males overexpressing both the growth hormone (GH) and its receptor (GHR)

et al. 2016

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Growth hormone (GH) transgenesis presents a high potential application in aquaculture. However, excess GH may have serious consequences due to pleiotropic actions. In order to study these effects in zebrafish (Danio rerio), two transgenic lines were developed. The first expresses GH ubiquitously and constitutively (F0104 line), while the second expresses the GH receptor in a muscle-specific manner (Myo-GHR line). Results from the F0104 line showed accelerated growth but increased reproductive difficulties, while Myo-GHR did not show the expected increase in muscle mass. Since the two lines appeared to display complementary characteristics, a double transgenic (GH/GHR) was created via crossing between them. This double transgenic displayed accelerated growth, however reproductive parameters remained uncertain. The objective of the present study was to determine the reproductive capacity of males of this new line, by evaluating sperm parameters, expression of spermatogenesis-related genes, and reproductive tests. Double transgenics showed a strong recovery in almost all sperm parameters analyzed when compared to the F0104 line. Gene expression analyses revealed that Anti-Müllerian Hormone gene (amh) appeared to be primarily responsible for this recovery. Reproductive tests showed that double transgenic males did not differ from non-transgenics. It is possible that GHR excess in the muscle tissues of double transgenics may have contributed to lower circulating GH levels and thus reduced the negative effects of this hormone with respect to reproduction. Therefore, it is clear that GH-transgenesis technology should take into account the need to obtain adequate levels of circulating hormone in order to achieve maximum growth with minimal negative side effects.

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Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch (Perca flavescens)

Cited by 6 publications

References 57 publications

Nanocarried antioxidants in freezing extenders for boar spermatozoa

Nanocarried antioxidants in freezing extenders for boar spermatozoa

Cryopreservation of sperm in annual fish Austrolebias minuano (Cyprinodontiformes; Rivulidae)

Reproductive parameters of double transgenic zebrafish (Danio rerio) males overexpressing both the growth hormone (GH) and its receptor (GHR)

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