Hyperactivated Motility of Frozen-Thawed Spermatozoa from Fertile and Subfertile Japanese Black Bulls Induced by Cyclic Adenosine 3',5'-Monophosphate Analogue, cBiMPS

Abstract

Abstract. This study investigated whether a cyclic adenosine 3',5'-monophosphate (cAMP) analogue, cBiMPS, could induce hyperactivated motility in frozen-thawed Japanese Black bull spermatozoa and compared the ability of spermatozoa to undergo hyperactivation between fertile and subfertile bulls. Frozen-thawed spermatozoa from 3 fertile and 2 subfertile bulls were washed, suspended in BO-Hepes medium and incubated in the presence of 0.1 mM cBiMPS for up to 4 h. At 1-h intervals, the spermatozoa were examined for hyperactivated motility. The proportions of spermatozoa showing a circular swimming pattern with asymmetrical flagellar beating and those showing whiplash beating of flagella to the total number of motile spermatozoa were expressed as C% and W%, respectively. The motile spermatozoa % was barely affected by treatment with cBiMPS or the fertility status of the sperm donor, although it gradually decreased in all sperm samples during the 4-h incubation. In the fertile bulls, the C% was 0% at 0 h of incubation but rapidly increased during the 1-h incubation with cBiMPS. It then decreased slightly towards 4 h concomitantly with a gradual increase in W% towards 4 h. In the subfertile bulls, however, the cBiMPS-induced increase of C% was delayed for 1-3 h, although the incubation time-related changes in mean W% were similar between the fertile and subfertile bulls. In the vehicle controls for cBiMPS, the C% and W% were 0% throughout incubation for all the samples examined. The results suggest that hyperactivation of the flagellum can be induced by the cAMP analogue, cBiMPS, in frozen-thawed Japanese Black bull spermatozoa and that induction of hyperactivation may serve as a useful tool for detection of functional abnormality of spermatozoa from subfertile Japanese Black bulls. Key words: Bull, Cyclic adenosine 3',5'-monophosphate (cAMP), Hyperactivation, Spermatozoa, Subfertility (J. Reprod. Dev. 56: [36][37][38][39][40] 2010) ammalian spermatozoa must acquire fertilizing ability prior to penetration of the oocyte, and this is generally called capacitation [1]. This change includes two kinds of processes: priming for the occurrence of the acrosome reaction, which is capacitation of the sperm head, and hyperactivated motility of the flagellum (hyperactivation). Capacitation, hyperactivation and the acrosome reaction are essential prerequisites for mammalian spermatozoa to fertilize eggs. Impairment of any of these functions might cause fertilization failure leading to possible infertility or subfertility. Such cases have also been reported in humans [2,3].Recent studies have revealed that in boar spermatozoa, hyperactivated motility is induced by cyclic adenosine 3',5'-monophosphate (cAMP), protein kinase A (PKA)-dependent tyrosine kinase and activation of phospholipase Cγ-protein kinase C signalling via SYK activation [4][5][6]. In bull spermatozoa, however, hyperactivated motility and capacitation may be regulated by different pathways [7], although Ca 2+ may be essential for both functions [8,9]. It has been...

Relationship of protein tyrosine phosphorylation state with tolerance to frozen storage and the potential to undergo cyclic AMP‐dependent hyperactivation in the spermatozoa of Japanese Black bulls

“…We previously showed that hyperactivation was induced in boar ejaculated spermatozoa by a long (90–180 min) incubation with cell‐permeable cyclic AMP (cAMP) analog “Sp‐5,6‐DCI‐cBiMPS” (cBiMPS) in vitro (Harayama and Miyake, 2006; Harayama and Nakamura, 2008). Similar effects of cBiMPS on the transition of motility to hyperactivation were recently observed in frozen‐stored spermatozoa of cattle (Japanese Black bulls) (Murase et al, 2010). Moreover, our previous findings (Harayama et al, 2004a, 2005) indicate that cBiMPS‐induced protein tyrosine phosphorylation is involved in the activation of spleen tyrosine kinase (SYK)‐phospholipase Cgamma1 (PLCgamma1) signaling cascades in the connecting piece of boar spermatozoa.…”

“…Moreover, we suggest that the immunodetection levels of tyrosine‐phosphorylated proteins in the sperm flagellum is related to the potential to undergo cAMP‐dependent hyperactivation. As low tolerance to frozen storage and low potential to undergo cAMP‐dependent hyperactivation cause AI subfertility (Bailey et al, 2000; Kuroda et al, 2007; Murase et al, 2010), we believe that the assessment of sperm protein tyrosine phosphorylation patterns is a valid tool for screening AI‐subfertile Japanese Black bulls.…”

Translocation of Tektin 3 to the equatorial segment of heads in bull spermatozoa exposed to dibutyryl cAMP and calyculin A

“…Phosphorylation of flagellar protein occurs during the transition to hyperactivation in mammalian spermatozoa (Si and Okuno, ; Baker et al, ; Kaneto et al, ). In vitro incubation of ejaculated porcine spermatozoa or frozen‐stored bovine spermatozoa (Japanese black bulls) with a cell‐permeable cyclic AMP (cAMP) analog induced hyperactivation, which might be mediated by cAMP‐induced protein phosphorylation in flagella (Harayama and Miyake, ; Harayama and Nakamura, ; Pons‐Rejraji et al, ; Murase et al, ). Calyculin A, a protein phosphatase inhibitor, also greatly enhanced cAMP‐triggered protein phosphorylation at serine/threonine and tyrosine residues in the connecting piece and also induced full hyperactivation of flagella, which is characterized by asymmetrical bending of the middle piece with a large amplitude (Mizuno et al, ).…”

“…induced hyperactivation, which might be mediated by cAMP-induced protein phosphorylation in flagella (Harayama and Miyake, 2006;Harayama and Nakamura, 2008;Pons-Rejraji et al, 2009;Murase et al, 2010). Calyculin A, a protein phosphatase inhibitor, also greatly enhanced cAMP-triggered protein phosphorylation at serine/threonine and tyrosine residues in the connecting piece and also induced full hyperactivation of flagella, which is characterized by asymmetrical bending of the middle piece with a large amplitude (Mizuno et al, 2015).…”

Improvement of Post-Thaw Sperm Kinematics and DNA Integrity of Cross-Bred Bovine Sperm by Incorporating DGC as Selection Method Prior to Cryopreservation

“…Findings suggest that compromise in the quality of cryopreserved sperm is likely due to synergistic effect of environmental and cyrobiological factors (Bailey et al, 2000). Previous studies have established that using sperm from heterogeneous populations for cryopreservation would later result in deteriorated post-thaw values for sperm kinematics and viability (Verstegen et al, 2002;Murase et al, 2010). Cooper et al (2010) defined high quality sperm as those exhibiting morphological and kinematic values above the minimal baseline values as suggested by the World Health Organization guidelines (Wakayama et al, 1998).…”