Mammalian sperm must undergo a set of structural and functional changes collectively termed as capacitation to ensure successful oocyte fertilization. However, can be compromised by cryopreservation procedures, which alter the proteome longevity sperm. To date, how protein induced could affect acquisition fertilizing potential remains unexplored. The present study investigated profile ram during in vitro before after elucidate impact on at molecular level. Fresh cryopreserved were incubated...

Rabbits constitute an interesting model to understand gamete interaction and test novel Artificial Reproductive Techniques, but in vitro fertilization (IVF) is particularly problematic this species. We have conducted a series of experiments develop consistent IVF technique. Initially, we checked viability, acrosome integrity, capacitation motility ejaculated sperm purified by density gradient incubated at different times three media: Tyrode's Albumin Lactate Pyruvate (TALP), human tubal...

Introduction and objective Cryopreservation of testicular tissues offers new possibilities to protect endangered species, genetically valuable individuals or even the fertility potential prepubertal who have died unexpectedly. However, use this technique still remains a challenge. In study, slow freezing vitrification tissue was investigated find out which cryopreservation method could better preserve viability DNA integrity germ cells in diverse wild species. Methods Testes were obtained...

Abstract Background Sperm chromatin structure provides valuable information for the prediction of male fertility and can be altered during different procedures. Previous studies have shown that sperm condensation decreased in vitro capacitation. Moreover, cryopreservation affect DNA integrity compaction. Objectives This study aimed to investigate dynamic modifications produced ram spermatozoa capacitation before after cryopreservation. Materials methods Chromatin decondensation (AB+),...

Abstract Background Recently, the metabolic pathways involved in energy production and role of aquaglyceroporins capacitation‐associated events have been studied humans mice. However, little is known about these ram spermatozoa. Objective The present study investigated bioenergetic aquaglyceroporin 3 variations during vitro capacitation In addition, differences testosterone levels between males were examined to determine their influence on capacitation‐like changes. Materials methods...

For the past two decades, there has been a growing interest in application of vitro embryo production (IVP) small ruminants such as sheep. To improve efficiency, large number abattoir-derived ovaries must be used, and long distances from laboratory are usually inevitable when adult animals used. In that scenario, prolonged sheep ovary transportation may negatively affect oocyte developmental competence. Here, we evaluated effect storage time (3, 5, 7, 9, 11 13 h) medium which they were...

Abstract This study aimed to compare the ability of sperm chromatin structure assay ( SCSA ® ) and Sperm‐Ovis‐Halomax detect DNA fragmentation in frozen‐thawed ram spermatozoa incubated under capacitating conditions synthetic oviductal fluid SOF supplemented with oestrous sheep serum ‐ ESS at multiple time points (0–240 min). Incubation had no significant effects on parameters while percentage fragmented measured by increased after 180 min incubation. In addition, correlation or agreement...

Creating germplasm banks of wild species, such as the Iberian red Deer (Cervus elaphus hispanicus) can be challenging. One main difficulties is obtention and cryopreservation good-quality reproductive cells when spermatozoa are obtained from epididymides after death. To avoid a loss seminal quality during transport, developing alternative methods for cooling freezing sperm samples under field conditions necessary. The objective this study was to evaluate effects different durations...

Abstract Sperm capacitation is a crucial step towards the acquisition of fertilizing capacity. Despite attempts to mimic in vivo situation, there still lack standardization vitro techniques. Bicarbonate and serum albumin (BSA) are routinely used, although controversial results reported regarding optimal concentration each compound. In addition, whether caffeine needed on media boar sperm remains be elucidated. Here, 18 commercial artificial insemination doses were used test different...

A major limiting factor for the development of in vitro embryo production (IVP) wild species, such as Iberian red deer, compared to livestock animals is poor availability and limited access biological material. Thus, use post-mortem ovaries from slaughtered represent a source oocytes large scale embryos needed research improve efficiency IVP. However, these are not developmentally competent their vivo counterparts. Moreover, usually obtained that have been transported long distances, which...

The heterogeneous nature of ejaculates highlights the relevance studying behavior different sperm subpopulations. Changes in motility and increase tyrosine phosphorylation are key events that usually occur during capacitation can be modified by cryopreservation process. However, relationship between both remains poorly defined throughout Fresh frozen-thawed spermatozoa were incubated capacitating (CAP) non-capacitating (NC) media up to 240 min. Sperm kinematics, mitochondrial activity...

To date, the underlying mechanisms by which cAMP modulators act during in vitro maturation to improve oocyte developmental competence are poorly understood. Here, we sought fill this knowledge gap evaluating use of phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) and adenylyl cyclase activator forskolin a culture period 2 h before (pre-IVM) on nuclear cytoplasmic features essential organelles, cumulus cells activity, potential sheep oocytes. Results showed that pre-IVM...

Incubation with estrous sheep serum (ESS) is required to induce in vitro capacitation of spermatozoa during fertilization small ruminants. However, the effect adding different concentrations media on quality resulting blastocysts has not yet been studied. Here, 298 oocytes were co-incubated capacitated either 10% or 2% ESS. There no differences between treatments cleavage (10% ESS: 63.81 ± 5.87% and 45.31 5.87%) blastocyst rates 20.83 2.12% 15.93 2.12%). Nonetheless, vitro-produced from ESS...

Assisted reproductive technologies are key to maintain genetic stocks of endangered wild species, such as the Iberian ibex (Capra pyrenaica). Due low availability ovaries, heterologous in vitro fertilization (IVF) with domestic goat hircus) oocytes constitutes an excellent alternative determine capacity sperm doses. The aim this study was optimize ibex-goat IVF procedures by testing two different media (TALP and SOF) whether estrous sheep serum (ESS) is required for fertilization. We found...

Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than ejaculated spermatozoa. This work examines effects lactoferrin (LF) and phosphoglycerate mutase 2 (PGAM2), which abundant in epididymal sperm ruminants, as additives Iberian ibex mouflon extenders. In addition, LF was added to a vitrification medium determine whether it also provided protection during cryopreservation testicular tissue. Epididymal samples testes were recovered post-mortem ibexes...

Contents Nowadays, the use of foetal calf serum ( FCS ) during in vitro embryo culture is very controversial. Whilst some authors have encouraged its use, others reject it because harmful effects. Although production red deer a promising assisted reproductive technique, still infancy and great effort needed to update protocols used. The aim this study was assess whether supplementation medium necessary produce blastocyst and, if so, when best time add terms quality. In blastocysts were...

An ejaculate is a mixture of sperm subpopulations (SP) with varying motility characteristics. Moreover, males high percentages fast and linear-moving have rates fertility (Ramón et al. 2003 Biol. Reprod. 89, 110). The objective was to assess dynamics, over time, SP capacitated red deer sperm. Thawed were selected 45/90% Percoll, diluted at 10 × 106 mL−1 in SOF plus 10% oestrous sheep serum incubated for 2 h 38.5°C under 5% CO2. Sperm assessed by computer-assisted semen analysis 1, 5, 15, 30,...