Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

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Red LED Light Acts on the Mitochondrial Electron Chain of Mammalian Sperm via Light-Time Exposure-Dependent Mechanisms

Cells ◽

10.3390/cells9122546 ◽

2020 ◽

Vol 9 (12) ◽

pp. 2546 ◽

Cited By ~ 1

Author(s):

Olga Blanco-Prieto ◽

Jaime Catalán ◽

Lina Trujillo-Rojas ◽

Alejandro Peña ◽

Maria Montserrat Rivera del Álamo ◽

...

Keyword(s):

Mitochondrial Function ◽

Red Light ◽

Maximal Effect ◽

O2 Consumption ◽

Led Light ◽

Intracellular Atp ◽

Mammalian Sperm ◽

Atp Levels ◽

Red Led ◽

Oligomycin A

This work analyzes the effects of red LED light on mammalian sperm mitochondrial function, using the pig as an animal model. Liquid-stored pig semen was stimulated with red-light for 1, 5 and 10 min in the presence or absence of oligomycin A, a specific inhibitor of mitochondrial ATP synthase, or carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), a specific disruptor of mitochondrial electron chain. Whereas exposure for 1 and 5 min significantly (p < 0.05) decreased total motility and intracellular ATP levels, irradiation for 10 min induced the opposite effect. Oligomycin A abolished the light-effects on intracellular ATP levels, O2 consumption and mitochondrial membrane potential, whereas compared to non-irradiated samples, FCCP significantly (p < 0.05) increased O2 consumption when sperm were irradiated for 1 min. Both oligomycin A and FCCP significantly (p < 0.05) decreased total motility. Red-light increased cytochrome c oxidase activity with a maximal effect after 5 min of irradiation, which was abolished by both oligomycin A and FCCP. In conclusion, red-light modulates sperm mitochondrial function via electron chain activity in an exposition, time-dependent manner.

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Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity

Andrologia ◽

10.1111/and.12301 ◽

2014 ◽

Vol 47 (5) ◽

pp. 545-552 ◽

Cited By ~ 40

Author(s):

M. N. Bucak ◽

M. B. Ataman ◽

N. Başpınar ◽

O. Uysal ◽

M. Taşpınar ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Parameters ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Bull Sperm

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Positive effects of trehalose and cysteine on ram sperm parameters

Veterinární Medicína ◽

10.17221/131/2016-vetmed ◽

2017 ◽

Vol 62 (No. 5) ◽

pp. 245-252 ◽

Cited By ~ 2

Author(s):

S. Gungor ◽

C. Ozturk ◽

AD Omur

Keyword(s):

Sperm Motility ◽

Sperm Parameters ◽

Control Group ◽

Mitochondrial Activity ◽

Sperm Viability ◽

Significant Protection ◽

Positive Effects ◽

Liquid Storage ◽

Acrosome Integrity ◽

Ram Sperm

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P < 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P < 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P < 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P < 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P < 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

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Epicatechin Provides Antioxidant Protection to Bovine Spermatozoa Subjected to Induced Oxidative Stress

Molecules ◽

10.3390/molecules24183226 ◽

2019 ◽

Vol 24 (18) ◽

pp. 3226 ◽

Cited By ~ 5

Author(s):

Eva Tvrda ◽

Peter Straka ◽

Drahomir Galbavy ◽

Peter Ivanic

Keyword(s):

Oxidative Damage ◽

Sperm Motility ◽

Reproductive Technologies ◽

Protein Carbonyls ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Ros Production ◽

Ros Scavenging ◽

Bovine Spermatozoa ◽

Dispersion Test

Epicatechin (EPI) is a natural flavonoid with antibacterial, anti-inflammatory and anti-cancer properties. Furthermore, the molecule exhibits powerful reactive oxygen species (ROS) scavenging and metal-chelating properties. In this study, we assessed the efficiency of EPI to reverse ROS-mediated alterations to the motility, viability, DNA integrity and oxidative profile of bovine spermatozoa. For the first experiment, spermatozoa were washed out of fresh semen and exposed to 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI and 100 μmol/L EPI in the presence of ferrous ascorbate (FeAA) during a 6 h in vitro culture. For the second experiment, the ejaculates were split into aliquots and cryopreserved with a commercial semen extender supplemented with 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI, 100 μmol/L EPI or containing no supplement. Sperm motility was assessed using the computer-aided sperm analysis and the cell viability was studied with the metabolic activity test. ROS production was quantified using luminometry, and DNA fragmentation was evaluated using the chromatin dispersion test. Cell lysates were prepared at the end of the culture in order to assess the concentration of protein carbonyls and malondialdehyde. Exposure to FeAA led to a significantly reduced sperm motility (p < 0.001), mitochondrial activity (p < 0.001), but increased the generation of ROS (p < 0.001), as well as oxidative damage to proteins (p < 0.001), DNA (p < 0.001) and lipids (p < 0.001). EPI supplementation, particularly at a concentration range of 50–100 μmol/L, resulted in higher preservation of the spermatozoa vitality (p < 0.001). Furthermore, 50–100 μmol/L EPI were significantly effective in the prevention of oxidative damage to sperm proteins (p < 0.001), lipids (p < 0.001) and DNA (p < 0.01 in relation to 50 μmol/L EPI; p < 0.001 with respect to 100 μmol/L EPI). In the case of the cryopreserved spermatozoa, the administration of 50–100 μmol/L EPI resulted in higher sperm motility (p < 0.001) and mitochondrial activity (p < 0.001). ROS production, the number of protein carbonyls, lipid peroxidation as well as oxidative DNA damage were found to be significantly decreased particularly in samples cryopreserved in the presence of 100 μmol/L EPI (p < 0.001). Our results suggest that EPI could behave as an effective antioxidant which may prevent oxidative insults to spermatozoa, and thus, preserve their vitality and functionality. Nevertheless, its potential to achieve higher fertilization rates in reproductive technologies needs to be validated.

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26 SPERM REPRODUCTIVE POTENTIAL OF FETAL FIBROBLAST CELLS ON A NUCLEAR TRANSFER - CLONED BULL AND ON AN ORIGINAL PATERNAL BULL: A COMPARATIVE ANALYSIS

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab26 ◽

2007 ◽

Vol 19 (1) ◽

pp. 131

Author(s):

S.-H. Bae ◽

D.-H. Kim ◽

S.-W. Kim ◽

G.-S. Im ◽

M.-J. Lee ◽

...

Keyword(s):

Sperm Motility ◽

Treated Group ◽

Flow Cytometer ◽

Mitochondrial Activity ◽

Computer Assisted ◽

Fibroblast Cells ◽

Sperm Viability ◽

Fetal Fibroblast ◽

Reproductive Ability ◽

Bull Sperm

Recently, cloning technology has produced newborn calves from fetal fibroblast cells and somatic cells. These cloned calves appear to be normal and healthy in terms of reproductive ability. Therefore, we undertook this study to compare the reproductive ability of cloned sperm from the clones themselves and sperm obtained from the frozen semen of the paternal bull (KPN-178). Two cloned bulls from fetal fibroblast cell nuclear transfers (Korean HanWoo Clone-38, 34 months old, and Clone-39, 30 months old) were selected. Sperm from the cloned groups (C-38 and C-39), cloned-38 fresh sperm, and sperm from the paternal bull (KPN-178) were analyzed for the following sperm reproductive characteristics: sperm motility, by computer-assisted sperm analysis (CASA); sperm mitochondrial activity, by flow cytometer after mitotracker staining; and sperm viability, by flow cytometer after staining with live/dead sperm viability kits, both PI and SYBR. Sperm motility was evaluated with Percoll-treated or non-Percoll-treated frozen-thawed spermatozoa. In the Percoll-treated group, total motility of the paternal bull sperm (KPN-178: 75.14%) was significantly lower than that of cloned sperm (C-38: 90.42%; C-39: 87.62%: C-38 fresh: 93.97%; P < 0.05). The non-Percoll-treated group showed different results compared to the Percoll results (P < 0.05), i.e. the total motility of C-38 (92.47%) and C-38 fresh (92.47%) sperm was significantly higher than that of C-39 (79.52%) and KPN-178 (78.41%) sperm. The mitochondrial activity staining rate of KPN-178 (58.22%) sperm was low compared to that of C-38 (88.65%), C-39 (89.45%), C-38 fresh (88.66%) sperm. The sperm viability tests showed similar results (P < 0.05) for all sperm groups: C-38 (PI: 42.53% and SYBR: 46.28%), C-39 (PI: 39.03% and SYBR: 66.18%), C-38 fresh (PI: 34.32% and SYBR: 57.10%), and KPN-178 (PI: 14.43% and SYBR: 47.86%). However, the PI staining of KPN-178 was lower than that of the other sperm groups. Next, in vitro fertilization (IVF) was investigated to compare the reproductive ability of C-38, C-39, KPN-178, and Control (normal bull) sperm. The results, development to the 2-cell stage (157/228, 68.90%; 85/116, 73.28%; 78/114, 68.42%; 61/79, 77.21%, respectively) and blastocyst rates (41/228, 18.0%; 35/116, 30.17%; 26/114, 22.81%; 20/79, 25.32%, respectively) were evaluated (P < 0.05). The IVF rates were not different among cloned bull, paternal bull, and normal bull sperm. A cloned female bovine was successfully impregnated by cloned bull sperm (C-38) using artificial insemination. The calf resulting from this prenancy showed no signs of phenotypical abnormalities. These results suggest that the physiology of cloned bulls and the quality of semen from cloned bulls show no evidence of any deleterious effects.

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In Vivo Monitoring of Intracellular ATP Levels inLeishmania donovani Promastigotes as a Rapid Method To Screen Drugs Targeting Bioenergetic Metabolism

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.4.1121-1125.2001 ◽

2001 ◽

Vol 45 (4) ◽

pp. 1121-1125 ◽

Cited By ~ 37

Author(s):

Juan Román Luque-Ortega ◽

Octavio Miguel Rivero-Lezcano ◽

Simon L. Croft ◽

Luis Rivas

Keyword(s):

Mitochondrial Membrane ◽

Leishmania Donovani ◽

Cell Number ◽

Rapid Screening ◽

Mitochondrial Activity ◽

Intracellular Atp ◽

Atp Levels ◽

Bioenergetic Metabolism ◽

Free Membrane

ABSTRACT A method for the rapid screening of drugs targeting the bioenergetic metabolism of Leishmania spp. was developed. The system is based on the monitoring of changes in the intracellular ATP levels of Leishmania donovani promastigotes that occur in vivo, as assessed by the luminescence produced by parasites transfected with a cytoplasmic form of Phothinus pyralisluciferase and incubated with free-membrane permeabled-luciferin analogued-luciferin–[1-(4,5-dimethoxy-2-nitrophenyl) ethyl ester]. A significant correlation was obtained between the rapid inhibition of luminescence with parasite proliferation and the dissipation of changes in mitochondrial membrane potential (ΔΨm) produced by buparvaquone or plumbagin, two leishmanicidal inhibitors of oxidative phosphorylation. To further validate this test, a screen of 14 standard leishmanicidal drugs, using a 50 μM cutoff, was carried out. Despite its semiquantitative properties and restriction to the promastigote stage, this test compares favorably with other bioenergetic parameters with respect to time and cell number requirements for the screening of drugs that affect mitochondrial activity.

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The Effects of Red Light on Mammalian Sperm Rely upon the Color of the Straw and the Medium Used

Animals ◽

10.3390/ani11010122 ◽

2021 ◽

Vol 11 (1) ◽

pp. 122

Author(s):

Jaime Catalán ◽

Iván Yánez-Ortiz ◽

Sabrina Gacem ◽

Marion Papas ◽

Sergi Bonet ◽

...

Keyword(s):

Plasma Membrane ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Sperm Motility ◽

Mitochondrial Membrane ◽

Red Light ◽

Mitochondrial Activity ◽

Dark Light ◽

Mammalian Sperm ◽

Intracellular Ros

Previous research has determined that irradiation of mammalian sperm with red light increases motility, mitochondrial activity, and fertilization capacity. In spite of this, no study has considered the potential influence of the color of the straw and the extender used. Therefore, this study tests the hypothesis that the response of mammalian sperm to red light is influenced by the color of the straw and the turbidity/composition of the extender. Using the horse as a model, 13 ejaculates from 13 stallions were split into two equal fractions, diluted with Kenney or Equiplus extender, and stored at 4 °C for 24 h. Thereafter, each diluted fraction was split into five equal aliquots and subsequently packed into 0.5-mL straws of red, blue, yellow, white, or transparent color. Straws were either nonirradiated (control) or irradiated with a light–dark–light pattern of 3–3–3 (i.e., light: 3 min, dark: 3 min; light: 3 min) prior to evaluating sperm motility, acrosome and plasma membrane integrity, mitochondrial membrane potential, and intracellular ROS and calcium levels. Our results showed that irradiation increased some motion variables, mitochondrial membrane potential, and intracellular ROS without affecting the integrities of the plasma membrane and acrosome. Remarkably, the extent of those changes varied with the color of the straw and the extender used; the effects of irradiation were more apparent when sperm were diluted with Equiplus extender and packed into red-colored straws or when samples were diluted with Kenney extender and packed into transparent straws. As the increase in sperm motility and intracellular ROS levels was parallel to that of mitochondrial activity, we suggest that the impact of red light on sperm function relies upon the specific rates of energy provided to the mitochondria, which, in turn, vary with the color of the straw and the turbidity/composition of the extender.

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Pengaruh Perlakuan Waktu dan Suhu Penyimpanan Dingin terhadap Mutu Kubis Bunga (Brassica Oleracea Var. Botrytis L.)

Jurnal BETA (Biosistem dan Teknik Pertanian) ◽

10.24843/jbeta.2020.v08.i01.p17 ◽

2019 ◽

Vol 8 (1) ◽

pp. 138

Author(s):

Chyntia Wulandari Eka Saputri ◽

I. A. Rina Pratiwi Pudja ◽

Pande Ketut Diah Kencana

Keyword(s):

Weight Loss ◽

Shelf Life ◽

Cold Storage ◽

Storage Time ◽

Consumption Rate ◽

Storage Temperature ◽

Treatment Time ◽

O2 Consumption ◽

Damage Level ◽

Four Levels

Tujuan dari penelitian ini adalah untuk menentukan waktu perlakuan optimal dan suhu penyimpanan dingin untuk mutu kubis bunga. Penelitian ini menggunakan rancangan acak lengkap (RAL) yang terdiri dari dua faktor, faktor pertama adalah suhu yang digunakan dan faktor kedua adalah waktu selama show case. Faktor pertama terdiri dari dua level, yaitu (P1): show case temperature 8oC, dan (P2): show case temperature 15oC dan tambah kontrol (P0). Faktor kedua terdiri dari empat level, yaitu (A0): penyimpanan selama 0 jam, (A1): penyimpanan selama 12 jam, (A2): penyimpanan selama 16 jam, (A3): penyimpanan selama 20 jam dan diulang untuk 3 kali ulangan. Kubis bunga sebagai kontrol disimpan pada suhu kamar (28 ± 1 ?). Parameter kualitas yang diamati dalam penelitian ini termasuk penurunan berat badan, tingkat konsumsi O2, warna (warna berbeda), uji organoleptik termasuk umur simpan dan tingkat kerusakan. Hasil penelitian menunjukkan parameter penurunan susut bobot, laju konsumsi O2, warna, umur simpan, tingkat kerusakan pada suhu perlakuan suhu terbaik adalah suhu 8 ? dan waktu penyimpanan 20 jam (P1A3).Kata kunci: kembang kol, waktu penyimpanan, suhu penyimpanan dingin The purpose of this study was to determine the optimal treatment time and cold storage temperature for the quality of cabbage flowers. This study uses a completely randomized design (CRD) consisting of two factors, the first factor is the temperature used and the second factor is the time during the showcase. The first factor consists of two levels, namely (P1): showcase temperature of 8oC, and (P2): showcase temperature of 15oC and added a control (P0). The second factor consists of four levels, namely (A0): storage for 0 hours, (A1): storage for 12 hours, (A2): storage for 16 hours, (A3): storage for 20 hours and repeated for 3 replications. Flower cabbage as control was stored at room temperature (28 ± 1 ?). The quality parameters observed in this study included weight loss, O2 consumption rate, color (color different), organoleptic tests including shelf life and damage level. The results showed the parameters of weight loss, O2 consumption rate, color, shelf life, damage rate at the best temperature of 8 ? and storage time of 20 hours (P1A3). Keywords: cauliflower, storage time, cold storage temperature

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