The potential of sperm bovine protamine as a protein marker of semen production and quality at the National Artificial Insemination Center of Indonesia

Abstract

Background and Aim: Protamine (PRM) is the major protein in the sperm nucleus and plays an essential role in its normal function. Moreover, PRM has great potential as a protein marker of semen production and quality. This study aimed to assess the potential of sperm bovine PRM as a protein marker of semen production and quality in bulls at the National Artificial Insemination (AI) Center of Indonesia. Materials and Methods: The semen production capacity of each bull was collected from frozen semen production data at the Singosari AI Center for 6 months, and was then divided into two groups (high and low). A total of 440 frozen semen straws from six Limousin (LIM), six Friesian Holstein (FH), six Peranakan Ongole (PO), and four Aceh bulls aged 4-5 years were used in the study. The frozen semen was used to measure the concentration of PRM1, PRM2, and PRM3 using the enzyme immunoassay method. The frozen semen was also used to assess the quality of the semen, including progressive motility (PM) through computer-assisted semen analysis, sperm viability through eosin–nigrosin analysis, and the DNA fragmentation index through Acridine Orange staining. Results: PRM1 was significantly higher in all bull breeds included in the study (p<0.00), followed by PRM2 (p<0.00) and PRM3 (p<0.00). PRM1 significantly affected semen production in LIM, FH, PO, and Aceh bulls (p<0.05). Moreover, PRM2 significantly affected semen production only in FH and Aceh bulls (p<0.05), whereas PRM3 affected this parameter in PO and Aceh bulls exclusively (p<0.05). Consistently and significantly, PRM1 was positively correlated with the PM and viability of sperm and negatively associated with its DNA fragmentation in LIM, FH, PO, and Aceh bulls (p<0.05; p<0.01). The correlation analysis between PRM2 and PRM3 and semen quality parameters varied across all bull breeds; some were positively and negatively correlated (p<0.05; p<0.01), and some were not correlated at all. Conclusion: PRM1 has excellent potential as a protein marker of semen production and quality in bulls at the National AI Center of Indonesia.

PRM1 Gene Expression and Its Protein Abundance in Frozen-Thawed Spermatozoa as Potential Fertility Markers in Breeding Bulls

“…After that, 10 µL of thawed sperm were dropped onto a slide and covered with a coverslip. Finally, four fields were evaluated with a range of 50–250 sperm cells automatically counted using Sperm Vision (Minitube, Tiefenbach, Germany) [ 20 ].…”

“…Each frozen sperm was thawed in a 37 °C water bath for 30 s, washed twice with PBS, and pelleted (about 25 × 10 6 sperm cells/mL) by centrifugation at 700× g for 15 min. The amount of PRM1 protein was then measured in sperm pellets using the enzyme immunoassay (EIA) technique, according to the manufacturer’s protocols (Bovine PRM1 , MyBioSource, Inc., San Diego, CA, USA) [ 20 ]. The prepared sperm pellets were briefly added to the appropriate wells.…”

“…At the spermatid round, histone and non-histone proteins are substituted by transitional proteins [ 16 ], which are withdrawn from the compacted chromatin of elongated spermatids and replaced by PRM until sperm development [ 15 , 16 , 17 ]. The alternation process produces highly dense chromatin, which is a basis of germ cell maturation and unique to haploid cells [ 17 , 18 , 19 , 20 , 21 ]. In bulls [ 20 ], the PRM1 gene is essentially critical in normal sperm functioning.…”

“…The alternation process produces highly dense chromatin, which is a basis of germ cell maturation and unique to haploid cells [ 17 , 18 , 19 , 20 , 21 ]. In bulls [ 20 ], the PRM1 gene is essentially critical in normal sperm functioning. However, PRM2 and PRM3 genes have been expressed in bovine testes [ 22 ].…”

“…It reportedly plays an essential role in determining the semen quality of Frieswal crossbred bulls (HF X Sahiwal) [ 27 ]. Our previous study also showed PRM1 as the protein with the most potential determinant of semen quality of various bulls in Indonesia compared to PRM2 and PRM3 variants [ 20 ]. However, studies that prove the potential of PRM1 as a biomarker of bull fertility used for AI in Indonesia have not been reported.…”