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Effect of tempol and straw size on rooster sperm quality and fertility after post-thawing.

ABSTRACT: The aim of the present study was to investigate the effects of tempol and straw size on rooster sperm post-thaw quality and fertility. Rooster semen was cryopreserved in Lake extender containing 0 (control), 5, 10, 15 and 20 μM tempol (in two different straw size, 0.25 and 0.5). The percentage of total and progressive sperm motility, VAP and VSL increased in the 10 µM tempol group. Moreover, 10 µM tempol led to lower ROS compared to other groups. The lowest percentage of apoptotic-like changes was detected when the extender was treated with 10 µM of tempol. The minimum ROS was observed in the group treated with 0.5 straw size. Straw size did not have any significantly effect on GPx and SOD activities and TAC of frozen-thawed sperm. The highest significant percentage of fertility and hatching rate were observed in 10 µM of tempol. The results of the present study showed that supplementation of the Lake cryopreservation medium with 10 µM tempol improved cryo-survival. Also, the results of the present study suggested that Lake cryopreservation medium with 0.5-ml straw may perhaps be an appropriate method to improve the quality and fertility post-thawed rooster sperm.

SUBMITTER: Najafi A

PROVIDER: S-EPMC9288470 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Effect of tempol and straw size on rooster sperm quality and fertility after post-thawing.

Najafi Abouzar A Mehdipour Mahdieh M Mohammadi Hossein H Mehdipour Zohreh Z Khorrami Behzad B Nazari Mahdi M

Scientific reports 20220716 1

The aim of the present study was to investigate the effects of tempol and straw size on rooster sperm post-thaw quality and fertility. Rooster semen was cryopreserved in Lake extender containing 0 (control), 5, 10, 15 and 20 μM tempol (in two different straw size, 0.25 and 0.5). The percentage of total and progressive sperm motility, VAP and VSL increased in the 10 µM tempol group. Moreover, 10 µM tempol led to lower ROS compared to other groups. The lowest percentage of apoptotic-like changes w ...[more]

PMID: 35842559

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Project description:Freezing and thawing diminish sperm motility and fertility by disrupting the cholesterol balance in sperm plasma and organelle membranes. The aim of this study was to elucidate the mechanisms through which exogeneous cholesterol treatment enhances the quality of frozen-thawed bull sperm. The incorporation of cholesterol was investigated using boron-dipyrromethene (BODIPY)-cholesterol, and BODIPY signals were detected not only in the plasma membrane but also in the midpiece region immediately after thawing. The positive signal of cholesterol in the midpiece region was inhibited by a scavenger receptor class B Type I (SR-BI) inhibitor, block lipid transport 1 (BLT-1). To comprehend the role of exogenous cholesterol in the functions of the plasma membrane, propidium iodide (PI)/Annexin V and peanut agglutinin lectin (PNA) staining were performed. The results showed that treatment with exogenous cholesterol increased the number of acrosome-intact sperm and decreased the number of sperm with damage to the plasma membrane. Moreover, since BODIPY signals were also observed in the midpiece region, mitochondrial function was evaluated using a flux analyzer and a flow cytometer with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (JC-1) staining, revealing an increase in the number of sperm with high-mitochondrial activity and oxygen consumption. Finally, to assess sperm fertility, computer-assisted sperm analysis (CASA) and IVF were carried out. Sperm velocities and fertilization rates in IVF were significantly enhanced by the addition of cholesterol just after thawing. Thus, the treatment with cholesterol after thawing protected the plasma membrane from the stress of thawing and maintained mitochondrial function, thereby preserving the fertilization ability of frozen-thawed bull sperm for conventional IVF and artificial insemination (AI). Therefore, the application of cholesterol just after thawing is a promising option for improving the fertility of frozen-thawed sperm.

Project description:BackgroundTechniques for sperm cryopreservation have exhibited their potential in male fertility preservation. The use of frozen-thawed sperm in in vitro fertilization (IVF) cycles is widespread today. However, many studies reported that cryopreservation might have adverse effects on sperm DNA integrity, motility, and fertilization, probably due to cold shock, intra- and extracellular ice crystals, and excess reactive oxygen species (ROS). Studies suggested that freezing and thawing impaired sperm viability and might adversely affect subsequent fertilization and pregnancy outcomes. The potential damage to fertilization and subsequent embryonic development and offspring health raises the concern on sperm cryopreservation. However, the above mentioned studies are limited to intracytoplasmic sperm injection (ICSI) cycles, while IVF is a more natural and patient-friendly method. IVF requires a higher quality of sperm than ICSI. However, the effect of freezing and thawing on sperm used for IVF remains unknown. Therefore, we aim to investigate the effect of freezing and thawing on ejaculated sperm and subsequent pregnancy and neonatal outcomes in IVF.MethodsThis retrospective cohort study at a tertiary-care academic medical center included 447 women who used paternal frozen-thawed ejaculated sperm and 31,039 women who used paternal freshly ejaculated sperm for IVF and underwent frozen-thawed blastocyst transfer from January 2011 to September 2021. To balance the baseline characteristics of the two groups, patients using frozen sperm were matched with control groups using a propensity score matching algorithm with a ratio of 1:3.ResultsAlthough sperm motility decreased from 82.04% to 75.70% (P < 0.001) after the freezing-thawing process, the fertilization rate (68.27% for frozen sperm and 67.54% for fresh sperm), number of viable embryos (1.98 and 2.16), clinical pregnancy rate (44.7% and 51.8%), and live birth rate (40.3% and 42.4%) were comparable between the two groups (all P > 0.05). For neonatal outcomes, no between-group differences were observed in offspring gender, gestational age, birthweight, and the rate of preterm birth (21.7% and 12.9%), low birthweight neonates (19.2% and 16.0%), and birth defects (0.0% and 0.8%) (all P>0.05).ConclusionsFrozen-thawed sperm had lower sperm motility but resulted in comparable embryonic, pregnancy, and neonatal outcomes versus fresh sperm in IVF cycles.

Dataset Information

Effect of tempol and straw size on rooster sperm quality and fertility after post-thawing.

Publications

Effect of tempol and straw size on rooster sperm quality and fertility after post-thawing.

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