Motility Parameters and Intracellular ATP Content of Rabbit Spermatozoa Stored for 3 Days at 15°C

Abstract

The effect of semen storage duration on motility parameters and ATP content of rabbit spermatozoa were investigated. Ejaculates were collected from 9 New Zealand White male rabbits and diluted with a commercial rabbit semen extender Galap. Semen was stored at 15 degrees C for 3 days. On each day of storage sperm motility and intracellular ATP content were evaluated. Sperm motility parameters were assessed using the computer-assisted sperm analysis (CASA) system and ATP content using the bioluminescence method. The time of storage had a significant effect on sperm motility parameters (except straight-line velocity) and ATP content. A significant correlation was observed between motility parameters and sperm ATP content. The motility parameters most strongly correlated with ATP content were total motile spermatozoa (r = 0.6364), progressively motile spermatozoa (r = 0.529), amplitude of lateral head displacement (r = 0.4178), curvilinear velocity (r = 0.4111) and average path velocity (r = 0.3743). Results show that motility parameters determined using the CASA system and intracellular ATP content are sensitive indicators of sperm quality during in vitro storage and may be useful for estimation of in vivo fertilizing ability of rabbit semen.

Motility, mitochondrial membrane potential and ATP content of rabbit spermatozoa stored in extender supplemented with GnRH analogue [des-Gly10, D-Ala6]-LH-RH ethylamide

“…However, in both groups (Control and GnRH) significant changes in motility parameters (except VSL) and percentage of spermatozoa showing high MMP and ATP content were observed throughout 3 days of storage. According to our previous studies (Gogol andWierzchoś-Hilczer 2009, Gogol 2013) the most sensitive parameters of semen quality were motility (especially TMOT and PMOT) and ATP content of the spermatozoa. A small (but statistically significant) decrease in the percentage of spermatozoa with high MMP (7.6% in Control and 6.2% in GnRH group) may suggest that this parameter is less suitable for estimating the in vitro quality of stored rabbit spermatozoa.…”

“…Moreover, MMP and ATP content of spermatozoa, both considered as sensitive indicators of energy metabolism and motility (Marchetti et al 2004, Gogol 2013, were determined. The results show that addition of the GnRH analogue to the extender did not have any significant effect on sperm function, as measured by motility parameters, MMP and ATP content.…”

“…The adenosine triphosphate (ATP) from spermatozoa cells was determined using the ViaLight Plus kit according to the manufacturer's instructions (Cambrex Bio Science Rockland, Inc., USA) (Gogol 2013). Prior to assay, samples composed of 10 μL of diluted semen were mixed with 100 μL Cell Lysis Reagent and incubated at room temperature for 5 minutes to extract ATP from cells.…”

“…Sperm motility parameters were assessed using a computer-assisted sperm analysis (CASA) system (Sperm Class Analyzer, S.C.A V5.1, Microptic, Barcelona, Spain) (Gogol 2013). Before assessment samples of semen were incubated at 37 o C for 30 minutes in a water bath.…”

Evaluation of effectiveness of an innovative semen extender (Formula®) comparing with a traditional extender (Lepus®) for artificial insemination in rabbits does

“…The availability of semen that can be stored for longer periods would allow an extension of the interval between semen collection and insemination of the females (Di Iorio et al 2014). Thus, enhancing AI performance in case of farms without male located far away from semen collection centres (Gogol, 2013). Previous researches showed that the fertilizing ability of rabbit spermatozoa was preserved for up to 48 h using chilled storage techniques (Roca et al 2000;L opez-Gatius et al 2005;Rosato & Iaffaldano 2011), thus enabling the transportation of superior genetics between nearby farms.…”