Standardization of egg activation and fertilization in sterlet (Acipenser ruthenus)

Vechtova

Füssy

et al. 2021

IJMS

Self Cite

The purpose of the current study was to analyze phenotypic and functional characteristics of common carp (Cyprinus carpio) spermatozoa during in vitro aging and to investigate whether global DNA methylation is affected by sperm aging. Milt was collected from five individual males, stored in vitro on ice in a refrigerator for up to 96 h post stripping (HPS) and used to fertilize eggs with intervals of 1, 24 and 96 h. Computer-assisted sperm analysis and a S3e Cell Sorter was employed to determine the spermatozoa phenotypic characteristics (motility, velocity, concentration and viability). In addition, pH and osmolality of the seminal fluid and the capacity of the spermatozoa to fertilize, hatching rate and health of the resulting embryos were examined at different aging times. Whole-genome bisulfite sequencing was used to compare the global and gene-specific DNA methylation in fresh and aged spermatozoa. The results demonstrated that spermatozoa aging in common carp significantly affects their performance and thus the success of artificial fertilization. The methylation level at the cytosine-phosphate-guanine (CpG) sites increased significantly with 24 HPS spermatozoa compared to the fresh group at 1 HPS and then decreased significantly at 96 HPS. A more detailed investigation of gene specific differences in the DNA methylation was hindered by incomplete annotation of the C. carpio genome in the public databases.

Section: Methodsmentioning

confidence: 99%

Changes in Phenotypes and DNA Methylation of In Vitro Aging Sperm in Common Carp Cyprinus carpio

Vechtova

Füssy

et al. 2021

IJMS

Self Cite

“…In the test tube, the E400 extender provided the best hatchability. An optimum ratio of spermatozoa: egg is usually thought to be a key factor in in vitro fertilization [60], but the volume of activation water and number of eggs during fertilization are very important and must always be taken into account [61], as is also evident in the present study. Moreover, the test tube conditions also mimic more accurately the natural reproductive conditions that aquatic animals have adopted through long-term evolution processes [62].…”

Section: Fertilization and Swim-up Larvae Ratesmentioning

confidence: 63%

Optimization of Sperm Management and Fertilization in Zebrafish (Danio rerio (Hamilton))

Franěk

Rodina

et al. 2021

Animals

Self Cite

The aim of the present study was to investigate the spontaneous motility of spermatozoa and to optimize sperm collection, short-term sperm storage, and fertilization in zebrafish Danio rerio. The movement of spermatozoon in water was propagated along the flagellum at 16 s after sperm activation then damped from the end of the flagellum for 35 s and fully disappeared at 61 s after activation. For artificial fertilization, milt must be added to an immobilizing solution, which stops the movement of sperm and keeps the sperm motionless until fertilization. E400 and Kurokura as isotonic solutions were shown to be suitable extenders to store sperm for fertilization for 6 h. E400 stored sperm for 12 h at 0–2 °C. Sperm motility decreased only to 36% at 12 h post stripping for the E400 extender and to 19% for the Kurokura extender. To achieve an optimal level of fertilization and swim-up larvae rates, a test tube with a well-defined amount of 6,000,000 spermatozoa in E400 extender per 100 eggs and 100 µL of activation solution has proven to be more successful than using a Petri dish. The highest fertilization and swim-up larvae rates reached 80% and 40–60%, respectively, with milt stored for 1.5 h in the E400 extender at 0–2 °C.

“…Generally, high motility and velocity indicated better sperm quality (Gage et al 2004). As a key variable of sperm quality, percentage of sperm motility was recognized as an important variable for successful fertilization (Linhart et al 2000(Linhart et al , 2020bRurangwa et al 2001Rurangwa et al , 2004Gallego et al 2013). It should also be emphasized that fast motile spermatozoa are very important for fertilization (Cheng et al 2021b).…”

Section: Aging Sperm In Vivomentioning

confidence: 99%

In vivo and in vitro aging of common carp Cyprinus carpio sperm after multiple hormonal application and stripping of males

Zhang

Linhartová

et al. 2022

Fish Physiol Biochem

Self Cite

The present study on common carp (Cyprinus carpio L.) sperm was designed to evaluate changes in sperm phenotypic variables during multiple sperm stripping with sperm storage: a) in vivo and b) in vitro. Similar males were multiple injected with carp pituitary (CP) 3 times 3 days apart. Sperm were stored in vivo in the body cavity for 0.5 days (fresh sperm) and 3 days (old sperm) after CP application, then sperm were collected and diluted with a carp extender 1:1 and stored in vitro on ice for 0, 3 and 6 days. In general, fresh sperm from the rst stripping had slightly better quality and quantity than old sperm from the second and third stripping, especially in the phenotypic parameters of number of total spermatozoa and number of total motile spermatozoa (P < 0.05). The highest kinetic and quantitative sperm variables were obtained in fresh and old sperm just after sperm collection at 0 days and then they decreased during in vitro sperm storage up to 6 days (P < 0.05). The fertilization, hatching and malformation rates from fresh sperm were similar compared with the old sperm. From the rst stripping, fresh sperm, even stored for 6 days in vitro, showed fertility, hatching and malformations at 92.5%, 91.5% and 1.3%, respectively. Multiple hormonal treatments with multiple male stripping together with 0.5 days of in vivo sperm storage followed by 6 days of in vitro storage are methods that can be recommended for use in common carp aquaculture.