Viability assessment of primary growth oocytes following ovarian tissue vitrification of neotropical teleost pacu (Piaractus mesopotamicus)

Marques, Lis Santos; Fossati, Ana Amélia Nunes; Leal, Martinha S.; Rodrigues, Rômulo Batista; Bombardelli, Robie Allan; Streit, Danilo Pedro

doi:10.1016/j.cryobiol.2018.03.009

Cited by 7 publications

(2 citation statements)

References 16 publications

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“…Thus, the concomitant utilization of these analyses with metabolic evaluation assays is imperative, given their demonstrated heightened sensitivity compared to isolated approaches such as the trypan blue staining method [28]. Remarkably, in oocytes, Marques et al (2018) [29] observed that the MTT assay exhibited enhanced sensitivity for distinguishing between treatments with higher viability than those with lower viability. This phenomenon may be attributed to the elevated mitochondrial concentration inherent to oocytes during speci c developmental stages.…”

Section: Discussionmentioning

confidence: 99%

Understanding rigor mortis impacts on fish gamete viability

Dantas,

Marques,

Freitas

et al. 2023

Preprint

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This study aimed to evaluate the viability of reproductive gametes in zebrafish (Danio rerio), at different rigor mortis stages. Viability assessments were conducted on oocytes at various developmental stages using LIVE/DEAD and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. For sperm evaluation, both kinetic (CASA) and morphological assessments (Rose Bengal staining) were performed. Results demonstrated that rigor mortis progression significantly impacted oocyte viability during post-rigor stages, with the following viability rates: pre-rigor (70.43 ± 12.31%), fresh/control (46.43 ± 12.54%), post-rigor (27.62 ± 22.29%), and rigor mortis (comparable to fresh/control). Conversely, sperm kinetics exhibited nuanced responses to the rigor mortis stages, with specific parameters showing sensitivity, whereas the others remained relatively stable. Sperm motility was higher in the fresh/control (63.23 ± 19.03%) and pre-rigor (58.96 ± 14.38%) compared to the post-rigor group (3.34 ± 4.65%). This study highlights the significance of the pre-rigor for successful gamete collection and preservation. These findings provide valuable insights for conservation efforts and optimization of genetic resource management for endangered fish species. This study aimed to develop effective assistive reproductive techniques by elucidating the interplay between rigor mortis and gamete quality, contributing to the broader goals of species conservation and maintenance of genetic diversity in fish populations.

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Section: Discussionmentioning

confidence: 99%

Understanding rigor mortis impacts on fish gamete viability

Dantas,

Marques,

Freitas

et al. 2023

Preprint

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“…These data show that immature follicles are more likely to survive after vitrification than follicles at more advanced stages of maturation (III, IV and V). The same vitrification protocol was applied to Piaractus mesopotamicus and the immature follicles showed 70% of viability using histological analysis and two staining protocols (trypan blue and fluorescein diacetate combined with propidium iodide) 13 .…”

Section: Introductionmentioning

confidence: 99%

Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue

Marques

Fossati

Rodrigues

et al. 2019

Sci Rep

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The aim of the present study was to compare the efficiency of vitrification and slow freezing techniques for the cryopreservation of zebrafish ovarian tissue containing immature follicles. In Experiment 1, assessment of cell membrane integrity by trypan blue exclusion staining was used to select the best cryoprotectant solution for each cryopreservation method. Primary growth (PG) oocytes showed the best percentage of membrane integrity (63.5 ± 2.99%) when SF4 solution (2 M methanol + 0.1 M trehalose + 10% egg yolk solution) was employed. The vitrification solution, which presented the highest membrane integrity (V2; 1.5 M methanol + 5.5 M Me2SO + 0.5 M sucrose + 10% egg yolk solution) was selected for Experiment 2. Experiment 2 aimed to compare the vitrification and slow freezing techniques in the following parameters: morphology, oxidative stress, mitochondrial activity, and DNA damage. Frozen ovarian tissue showed higher ROS levels and lower mitochondrial activity than vitrified ovarian tissue. Ultrastructural observations of frozen PG oocytes showed rupture of the plasma membrane, loss of intracellular contents and a large number of damaged mitochondria, while vitrified PG oocytes had intact mitochondria and cell plasma membranes. We conclude that vitrification may be more effective than slow freezing for the cryopreservation of zebrafish ovarian tissue.

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