Effects of hypothermic storage on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and plasma membrane integrity in striped bass (Morone saxatilis) sperm

Guthrie, Helen A.; Welch, G. Rickey; Theisen, Daniel D.; Woods, L. Curry

doi:10.1016/j.theriogenology.2010.10.037

Cited by 24 publications

(11 citation statements)

References 34 publications

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“…Nearly 100% plasma membrane integrity of the freshly collected fish semen was showed earlier [19] and the high live-cell ratio was also observed in another species like carp [13]; striped bass [9,10]; and zebrafish [11]. The plasma membrane lesion is a late end point of necrotic cell death.…”

Section: Discussionmentioning

confidence: 67%

Effects of high ambient temperature on fish sperm plasma membrane integrity and mitochondrial activity — A flow cytometric study

Nagy

Kakasi

Pál

et al. 2016

Acta Biologica Hungarica

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Local extreme climatic conditions occurring as a result of global climate change may interfere with the reproduction of animals. In the present study fish spermatozoa were incubated at different temperatures (20, 25, 30 and 40 °C) for 10 and 30 minutes, respectively and plasma membrane integrity and mitochondrial membrane potential changes were evaluated with flow cytometry using SYBR-14/PI and Mitotracker Deep Red FM fluorescent dyes. No significant differences were found in plasma membrane integrity at either incubation temperatures or time points. Mitotracker Deep Red FM histogram profiles indicating mitochondrial activity showed significant (p < 0.001) alterations in all cases of higher (25, 30 and 40 °C) temperature treatments as compared to the samples incubated at 20 °C. Our results indicate that fish spermatozoa exposed to high temperatures suffer sublethal damage that cannot be detected with conventional, vital staining techniques.

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Section: Discussionmentioning

confidence: 67%

Effects of high ambient temperature on fish sperm plasma membrane integrity and mitochondrial activity — A flow cytometric study

Nagy

Kakasi

Pál

et al. 2016

Acta Biologica Hungarica

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“…We observed that phosphatase inhibition on ice as described by Guthrie et al (2011) in striped bass spermatozoa and the use of a Tricine boiling buffer (Ford and Leach, 1998) yielded highest ATP concentration regardless whether ATP was extracted from fresh or frozen samples. Therefore, ice conditions effectively prevented ATP degradation during phosphatase inhibitor treatment.…”

Section: Discussionmentioning

confidence: 99%

Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

et al. 2016

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Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

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“…The approximate percentage of cell motility was estimated as described previously. [50][51][52][53] The movements of rohu spermatozoa were recorded as movies (400-500 frames/minute) for 1 minute. The original movies were processed using Movie-maker software.…”

Section: Sds Page and Western Blot Analysismentioning

confidence: 99%

Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility

et al. 2013

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Effects of hypothermic storage on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and plasma membrane integrity in striped bass (Morone saxatilis) sperm

Cited by 24 publications

References 34 publications

Effects of high ambient temperature on fish sperm plasma membrane integrity and mitochondrial activity — A flow cytometric study

Effects of high ambient temperature on fish sperm plasma membrane integrity and mitochondrial activity — A flow cytometric study

Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility

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