Ascorbic Acid, Catalase and Chlorpromazine Reduce Cryopreservation-induced Damages to Crossbred Bull Spermatozoa

Abstract

The present study evaluated the effectiveness of ascorbic acid, catalase, chlorpromazine and their combinations in reducing the cryodamages to crossbred bull (Bos taurus x Bos indicus) spermatozoa. A total of 32 ejaculates (eight each from four bulls) were diluted in Tris-citric acid-fructose-egg yolk-glycerol extender. Each ejaculate was split into six parts (five treatment and one control). Treatment groups included 10 mm ascorbic acid, 0.1 mm chlorpromazine, 200 IU/ml catalase, 10 mm ascorbic acid + 0.1 mm chlorpromazine or 200 IU/ml catalase + 0.1 mm chlorpromazine in the extender. Fluorescent probes (Fluorescein isothiocyanate--Pisum sativum agglutinin + Propidium iodide) were used for the assessment of spermatozoa viability and acrosomal status. The proportion of acrosome intact live (AIL), acrosome intact dead, acrosome reacted live and acrosome reacted dead sperm was assessed in fresh, equilibrated and frozen-thawed semen. The functional status of the sperm was assessed using hypo-osmotic sperm swelling test (HOSST). Activities of acrosin and hyaluronidase enzyme were also determined. Lipid peroxidation level was assayed based on the melonaldehyde (MDA) production. In cryopreserved semen, the values of AIL spermatozoa, HOSST response, hyaluronidase and acrosin activity were reduced by 53%, 47%, 34% and 54%, respectively from their initial values in fresh semen. However, MDA level was threefold higher in the frozen-thawed sperm compared with fresh sperm. Significant (p < 0.05) improvement in motility, viability, HOSST response, retention of hyaluonidase and acrosin and reduction in MDA was recorded in ascorbic acid, catalase, ascorbic acid + chlorpromazine and catalase + chlorpromazine incorporated groups. The percentage of AIL sperm was significantly (p < 0.05) higher in ascorbic acid, catalase and ascorbic acid + chlorpromazine incorporated groups compared with the control. Chlorpromazine alone did not improve the post-thaw semen quality but when combined with either ascorbic acid or catalase, improvement in semen quality was noticed. It was inferred that incorporation of ascorbic acid, catalase and ascorbic acid + chlorpromazine in semen extender improved the post-thaw semen quality in crossbred bulls.

Effects of reduced glutathione on acrosin activity in frozen–thawed boar spermatozoa

“…Following freeze-thawing, there was a marked reduction in acrosin activity in both GFE and PFE, which was in agreement with the results of Pinart et al (2015). In fact, sperm cryopreservation is also known to significantly reduce acrosin activity in other mammals, such as humans (Cross and Hanks 1991), bulls (Paudel et al 2010) and bucks (Sirat et al 1996). However, the decrease in acrosin activity following freeze-thawing has been interpreted in a different manner by authors conducting their work in different species.…”

“…Although no previous study has addressed this issue in cryopreserved spermatozoa from other mammalian species, different studies have evaluated the effects of other antioxidants. For example, Paudel et al (2010) showed that adding ascorbic acid, catalase and chlorpromazine to cryopreservation media not only reduced sperm cryodamage in bulls, but also better maintained their acrosin activity. In humans, Rao et al (2015) recently found that testicular hyperthermia increases oxidative stress and decreases acrosin activity.…”

Protective effects of ascorbic acid and vitamin E on antioxidant enzyme activity of freeze-thawed semen of Qinchuan bulls

“…Vc is another primary component of the antioxidant system. Vc can improve sperm viability, decrease the percentage of abnormality, and increase the activity of sperm motility (Dawson et al, 1992;Branco et al, 2010;Paudel et al, 2010).…”

Effect of Incorporation of Additives in Tris‐Based Egg Yolk Extender on Buffalo (Bubalus bubalis) Sperm Tyrosine Phosphorylation During Cryopreservation

“…This improves motility, viability and functional integrity of spermatozoa (Thiangtum et al. 2009; Paudel et al. 2010).…”