Analysis of the motility parameters of in vitro hyperactivated hamster spermatozoa

Abstract

Golden hamster cauda epididymal spermatozoa under in vitro capacitating conditions exhibited time-dependent transformation of motility pattern, and the frequency of occurrence of a particular motility type was found to be dependent on the depth of the motility chamber used. The nonhyperactivated spermatozoa with planar motility were the most predominant at 0 hr irrespective of the depth of the motility chamber. But spermatozoa with the helical motility pattern were not detectable up to 6 hr when the Makler chamber was used, whereas in both the slide chamber and cannula, by 2 hr such spermatozoa constituted 90% of the total spermatozoa. However, by 6 hr the hyperactivated circular moving spermatozoa were the predominant type in all the chambers. Sperm motility chamber depths were also found to effect the motility parameter values of hamster spermatozoa, but this effect was also found to be dependent on the type of motility. Increase in chamber depth did not alter any of the motility parameter values of spermatozoa with hatchet type of motility and only increased the amplitude of lateral head displacement (ALH) in planar type. But in spermatozoa with the helical type of motility, an increase in chamber depth increased the progressive velocity (VSL), path velocity (VAP), curvilinear velocity (VCL), straightness (STR), linearity (LIN), and ALH. In spermatozoa with the circular type of motility, an increase in VSL, VAP, VCL, and ALH was also observed, but STR and LIN decreased. The hyperactivated spermatozoa could be distinguished from the nonhyperactivated spermatozoa because the former were swimming in circles, had low progressive velocity, decreased straightness and linearity of path, and also exhibited an increase in the amplitude of lateral head displacement compared to the nonhyperactivated spermatozoa. Further, the spermatozoa with helical motility could be differentiated from the nonhyperactivated, circular, and hatchet spermatozoa in that they had the highest VSL, VAP, VCL, and ALH. Spermatozoa with hatchet movement were slow and exhibited very low STR and LIN. Thus the motility parameters could be used to distinguish the nonhyperactivated and hyperactivated distal cauda epididymal spermatozoa.

Implications of diversity in sperm size and function for sperm competition and fertility

“…Analyses were carried out with logtransformed variables. Data for straightline velocity (VSL) were obtained from the literature (Mortimer and Mortimer, 1990;Stachecki et al, 1993Stachecki et al, , 1994Shivaji et al, 1995;Mahonynet al, 1996;Moore and Akhondi, 1996;Trimeche et al, 1996Trimeche et al, , 1999Patil et al, 1998;van der Horst et al, 1999;Castellini et al, 2000Castellini et al, , 2006Jayaprakash et al, 2001;Kawaguchi et al, 2004;Lavara et al, 2005;Miró et al, 2005;Baumber and Meyers, 2006;Cox et al, 2006;Sloter et al, 2006;Aurich et al, 2007;Grzmil et al, 2007;Holt et al, 2007;Hung et al, 2007;Immler et al, 2007a;Rijsselaere et al, 2007) and our own unpublished observations. Data for total sperm length were obtained from Cummins and Woodall (1985), Cassinello et al (1998), Gage and Freckelton (2003) and Malo et al (2006 Pitnick et al, 2001).…”

Pentoxifylline-stimulated capacitation and acrosome reaction in hamster spermatozoa: involvement of intracellular signalling molecules

“…Using the 'play back' and 'zoom function', the following kinematic parameters were determined for 100-150 individual spermatozoa from each treatment: straight line velocity (VSL), average path velocity (VAP), beat cross frequency (BCF), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), straightness (STR) and linearity (LIN). As detailed elsewhere (Shivaji et al, 1995;Jayapraksh et al, 1997), the sort function of the analyser was used to determine the populations of spermatozoa exhibiting planar (VSL Ͼ80 µm/s, STR Ͼ80%, LIN Ͼ50%, ALH Ͻ20 µm), or helical or circular (VSL Ͼ30 µm/s, STR Ͻ80%, LIN Ͻ50%, ALH Ͼ15 µm) patterns of motility, by visualizing individual sperm tracks. Hyperactivated spermatozoa, defined as those with helical or circular motility patterns, were expressed as a percentage of the total sperm population.…”

Serine/threonine phosphorylation associated with hamster sperm hyperactivation

“…IN ORDER TO examine the successive changes in serine and threonine phosphorylation that are related to sperm motility, we prepared three suspensions of spermatozoa: immotile, activated and hyperactivated spermatozoa. The immotile spermatozoa were obtained by dilution of mature epididymal spermatozoa in pure water, 19 activation occurred after dilution of spermatozoa in m‐TALP (they moved progressively with a high beat frequency of approximately 10 Hz) 8,19 and hyperactivated spermatozoa were obtained after 3 h incubation in m‐TALP; they exhibited a beat frequency of approximately 10 Hz, large bend amplitude, whiplash and frenzied flagellar movements 4–6,8 . We therefore redefined the activation of spermatozoa as the change from immotile to active and hyperactivation as the change from active to hyperactive.…”

“…Activated spermatozoa can be prepared in vitro by adding activating factors, such as calcium and/or bicarbonate, and are moved progressively with a high‐beat frequency 1–3 . Capacitated spermatozoa are prepared in vitro by adding calcium, albumin, glucose, etc., and capacitation involves an acrosomal reaction in the sperm head, which is very important for fertilization, and hyperactivation of motility, 1 which comprises a large bend amplitude, whiplash and frenzied flagellar movements 4–6 …”

“…[1][2][3] Capacitated spermatozoa are prepared in vitro by adding calcium, albumin, glucose, etc., and capacitation involves an acrosomal reaction in the sperm head, which is very important for fertilization, and hyperactivation of motility, 1 which comprises a large bend amplitude, whiplash and frenzied flagellar movements. [4][5][6] The most recent understanding is that the changes in sperm motility are closely associated with phosphorylation of flagellar proteins, 3 particularly serine, threonine and tyrosine; for example, 175-, 93-, 44-, 40-, 38-, 20-kDa proteins have been detected in boar sperm, 7 90-, 80-, 62-, 48-kDa proteins in hamsters 8,9 and 190-, 110-, 94-, 43-55-, 35-, 18-kDa proteins in humans. 10 From these results, it has been assumed that tyrosine phosphorylation plays an especially important role in the hyperactivation of mammalian spermatozoa.…”