Protein profile and functionality of spermatozoa from two semen collection methods in Bali bulls

Markers for Sperm Freezability and Relevance of Transcriptome Studies in Semen Cryopreservation: A Review

“…Even though post-thaw sperm motility is a good indicator of viability, it is not always an accurate fertility predictor of an AI-semen dose [9]. Evaluations of sperm motility characteristics have been improved by the incorporation of the computer-assisted semen analysis (CASA) system, which measures several motility and motion parameters of spermatozoa that are closely related to fertility compared with subjective motility measurements [21][22][23][24]. Besides motility analysis, studies have conirmed that the velocity parameters, such as velocity straight line (VSL), velocity curvilinear (VCL), and velocity average path (VAP), are associated with the fertilizing capacity of frozen-thawed spermatozoa [21,24].…”

“…Post-thaw sperm PMI has been assessed with diferent luorescent membrane probes, such as the dual SYBR-14 and propidium iodide (PI) assay [25,28] or membrane-permeable substrate carboxyluorescein diacetate (CFDA), a nonspeciic esterase substrate [24]. The chlortetracycline (CTC) luorescence assay has been used to detect capacitation-like changes in frozen-thawed spermatozoa, which may compromise their fertilizing ability [6,10,[22][23][24]29]. Cryo-induced changes in the acrosome membrane integrity (AMI) have been monitored by speciic Giemsa-staining technique [25] or with luorescent dyes, such as luorescein isothiocyanate (FITC)-conjugated PNA (peanut agglutinin) or conjugated PSA (Pisum sativum agglutinin), known as plant lectins, which bind to glycoproteins in the outer acrosomal membrane [4,5,26,27].…”

“…Moreover, there is accumulating evidence indicating that the cryopreservation procedure induces apoptotic-like features in bovine spermatozoa, which appeared as ordered events during the freezing-thawing process, such as reduced mitochondrial membrane potential (MMP), increased caspase activation, and modiications in membrane permeability [6,21,24,27,28,30]. Additionally, post-thaw PMI has been assessed with the hypo-osmotic swelling (HOS) test, which evaluates the functional membrane integrity of the acrosome and tail regions when spermatozoa are exposed to hypoosmotic conditions [22,24,29].…”

“…Moreover, a fertilityassociated protein, osteopontin (OPN), an acidic glycoprotein occurring in the bovine SP, has been shown to induce capacitation and improve viability of spermatozoa by inhibiting the apoptotic pathways [39,41]. Bovine SP proteins, collectively known as binder of sperm (BSP) proteins (BSP1, BSP 3, and BSP 5), bind to choline phospholipids in the sperm plasma membrane and are implicated in semen freezability [22,39,41,42]. Recently, it has been conirmed that BSP1, one of the most abundantly expressed BSP proteins (representing approximately 25-47% of the total proteins in bovine SP), consists of four molecular forms that have varying cryoprotective efects on the bull spermatozoa [42].…”

“…Recently, it has been conirmed that BSP1, one of the most abundantly expressed BSP proteins (representing approximately 25-47% of the total proteins in bovine SP), consists of four molecular forms that have varying cryoprotective efects on the bull spermatozoa [42]. According to Sarsaii et al [22], approximately 52% of the SP protein spots detected after cryopreservation were represented by four major protein fractions with diferent molecular weights, and 10 proteins, identiied by mass spectrometry, were major bovine SP proteins. It is worth noting that two of these proteins, deined as phosphoglycerate kinase (PGK, 37-45 kDa) and phospholipase A2 (PLA2, 50-55 kDa), are implicated in glycolysis and the fertilization-associated processes, respectively [22].…”