Gene Expression Profile and Enzymatic Activities of Frozen Buck Sperm Supplemented with Melatonin in Cold and Hot Temperatures.

Abstract

This study was carried out to improve the freezability of buck semen using two different types of cryoprotectants supplemented with melatonin as antioxidant in cold and hot temperature of breeding season. Ejaculates from four mature Egyptian baladi bucks were pooled after collection. Semen was extended with Tris-fructose-citric containing egg yolk using glycerol and dimethyl sulfoxide supplemented with two doses of melatonin (10-6 M and 10-3 M) in addition to control group. Types of motility as well as velocity, enzymatic activity and expression profile of selected genes were measured. The results revealed that the progressive motility percentage was significantly higher in samples supplemented with low dose of melatonin (10-6 M) compared to high dose (10-3 M) in glycerol (74.4 versus 64.4) and Dimethyl Sulfoxide (DMSO) based extender (35.5 versus 32.9) in cold temperature. The same trend was found in samples cryopreserved with glycerol (75.1 versus 53.5) and DMSO (32.1 versus 22) in hot temperature. The results also demonstrated that CASA parameters (VAP and VCL) were significantly increased in low compared to high melatonin dose in glycerol based extender during cold and hot temperature. The activity of total antioxidant capacity (TAC) was significantly higher in samples supplemented with low (0.49 mM/L) than high melatonin dose (0.16 mM/L) in DMSO extender. CPT2, ATP5F1A and SOD2 genes were up regulated in glycerol based extender groups in cold temperature compared to other groups of this study. On the other hand, NFE2L2 gene was upregulated in groups cryopreserved with DMSO in hot temperature compared with all other experimental groups. Therefore, it could be concluded that the glycerol based extender in cold season supplemented with low dose of melatonin improved semen quality, antioxidant defense capacity and transcriptional profile, which may maintain the post-thaw fertilizing ability of buck semen.

Age-Dependent Computer-Assisted Sperm Analysis of Buck Semen Supplemented with Different Antioxidants

“…In line with the previous findings, melatoninsupplemented male goat semen showed enhanced total and progressive sperm motility values for CASA parameters [30]. It should be noted that the inclusion of antioxidants (L, M, and S) is necessary to maintain the viability and quality of semen following cryopreservation [27,28,29].…”

“…To maintain semen quality and viability following cryopreservation, antioxidant addition (L, M, and S) is necessary [27,28,29] . The present study showed an increase in total and progressive motility in semen groups supplemented with various antioxidants, which is consistent with earlier investigations.…”

“…However, Motility and fertilization potential have enhanced due to the cysteine addition of semen extender in buck semen [29] . Additionally, it was shown that adding melatonin to buck semen increased sperm motility and vitality after freezing [28].…”

Molecular and physiochemical evaluation of buck semen cryopreserved with antioxidants

“…Our results demonstrated up-regulation of two mitochondrial transcripts (CPT2 and ATP5F1A) that enhance metabolic activity of spermatozoa in semen groups supplemented with Cys, LC + M, LC + Cys and M + Cys. Similarly, up-regulation of these two genes was reported in ram semen that showed enhancement in post-thaw sperm motility (Dessouki et al, 2020). In a microarray study, transcripts associated with keeping haemostasis of mitochondrial membrane potential were upregulated in high-fertile bulls (Madeja et al, 2021;Saraf et al, 2021).…”

Antioxidant Activity and Oxidative Stress-Oriented Apoptosis Pathway in Saccharides Supplemented Cryopreserved Sperm of Pacific Abalone, Haliotis discus hannai

“…Cryopreservation process can also reduce mRNA expression levels of five vital antioxidant enzymes (CAT, Cu/Zn-SOD, Mn-SOD, GPx, and GR) in mammalian sperm [ 13 , 14 , 15 ]. Imbalance in antioxidant defense might be responsible for mitochondria-oriented apoptosis [ 16 , 17 , 18 ].…”