Ramifications of protease-based liquefaction of camel semen on physical, kinematic and surface glyco-pattern of cryopreserved spermatozoa

Rateb, Sherif A.; Monaco, Davide; El-Bahrawy, K. A.; Khalifa, Marwa A.; El-Hamid, Ibrahim Samir Abd; Kamel, Ahmed Mandouh; Accogli, Gianluca; Lacalandra, Giovanni Michele; Desantis, Salvatore

doi:10.1016/j.anireprosci.2019.106121

Cited by 8 publications

(6 citation statements)

References 38 publications

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“…According to this postulation, absence of such protein(s) due to semen processing for cryopreservation could contribute to occurrence of this phenomenon. Additionally, prevalent anomalies in sperm of protease‐liquefied semen, specifically in the acrosome region, could be another factor that contributed in incidence of head‐to‐head sperm agglutination in the present work due to the drastic effects of both enzymes on acrosomal glycan features of spermatozoa (Rateb et al., 2019).…”

Section: Discussionmentioning

confidence: 80%

“…On the other hand, protease‐treated semen specimens in the current work exhibited higher percentages of secondary sperm deformities, altered acrosome and sperm cell membrane disintegrity compared to that of control prior to purification. This deterioration in sperm criteria was mainly attributed with the proteolytic actions of both enzymes on sperm acrosome and cell membrane glycocalyx (Rateb et al 2019). The results also showed a dramatic reduction in sperm DNA integrity in all cryopreserved specimens prior to sperm purification (T 0 ).…”

Section: Discussionmentioning

confidence: 99%

“…In our previous work, we pointed out that both bromelain‐ and papain‐based semen liquefaction dramatically affected the glycocalyx pattern of cryopreserved camel spermatozoa. The lectin histochemistry investigation indicated presence of N‐ and O‐linked oligosaccharides in the acrosomal cap and tail of spermatozoa with the latter resulting in PNA‐reactivity in these regions of sperm cell (Rateb et al., 2019). In the present study, the dextran‐coated NPs were conjugated with the lectins PNA, LCA and Annexin A5 to specifically target various defective camel sperm glycans.…”

Section: Discussionmentioning

confidence: 99%

“…The current study was implemented at Artificial Insemination Lab., Mariout Research Station (Latitude 31°00'N; Longitude 29°47'E), Alexandria, Egypt. The cryopreserved camel semen doses were provided by the laboratory to implement this work as a direct succession to a previous study on dromedary semen liquefaction (Rateb et al 2019). The procedures of semen collection, processing and cryopreservation, as well as the protease‐based liquefaction technique, were extensively described in the aforementioned study.…”

Section: Methodsmentioning

confidence: 99%

“…Enzymatic liquefaction of camel semen, particularly by using proteolytic enzymes, has been considered the most effective for elimination of camel semen viscosity and improving motility and kinematics of spermatozoa (Kershaw‐Young et al., 2017; Mal et al., 2016; Monaco et al., 2016). Nevertheless, protease‐based semen liquefaction was recently reported to drastically affect sperm vitality, acrosome integrity and sperm cell surface glycocalyx pattern especially after exposure to the stress of freezing/thawing cycle (El‐Bahrawy et al., 2017; Rateb et al., 2019). This, in turn, has spurred the search for new approaches to enhance cryopreservation outcome of protease‐liquefied dromedary semen.…”

Section: Introductionmentioning

confidence: 99%

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Purification of cryopreserved camel spermatozoa following protease‐based semen liquefaction by lectin‐functionalized DNA‐defrag magnetic nanoparticles

Rateb

2020

Reprod Domestic Animals

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Although incorporating proteases into sperm medium is considered the most effective procedure to eliminate camel semen viscosity, it drastically affects viability, morpho‐functional properties and, hence, fertilization potential of spermatozoa. The present work aimed at evaluating adequacy of employing magnetic nanoparticles‐based sperm purification technique for eluting impaired and apoptotic camel spermatozoa from cryopreserved semen doses following protease‐based semen liquefaction. Thirty cryopreserved semen doses (50 x 106 sperm/straw) representing the following liquefaction treatments: control (untreated), 0.1 mg/ml papain or 5 U/ml bromelain were used (n = 10 straws per treatment). Immediately after thawing (38°C for 40 s), sperm concentration of each straw within treatment was readjusted to 15 x 106 sperm/mL by dilution in PBS (37°C). Sperm physical and cytological properties were then assessed (non‐purified semen). Thereafter, each specimen was subjected to lectin‐functionalized DNA‐defrag magnetic nanoparticles sperm purification, and the same sperm traits were re‐evaluated after undergoing purification (purified semen). Sperm DNA fragmentation level within each group, prior to and after magnetic nano‐purification, was also determined by fluorescent imaging. The results showed a dramatic improvement (p < .05) in post‐thaw motility (%), viability (%), normal sperm (%), intact acrosome (%) and HOST‐reacted (%) spermatozoa in protease‐liquefied semen following sperm magnetic nano‐purification. Additionally, the highest (p < .05) DNA fragmentation level was recorded in all cryopreserved semen groups prior to purification, whereas the lowest (p < .05) was observed in the protease‐treated specimens after magnetic nano‐purification. These results indicate that protease‐based semen liquefaction prior to cryopreservation in conjunction with magnetic nano‐purification post‐thawing holds potential for reducing the proportion of damaged and dead spermatozoa, hence improving camel sperm fertilization competence.

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Section: Discussionmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%