DissertationChairdin Dwi Nugraha, S.Pt.,M.Pt.: Analysis of Semen Productivity of Bali Cattle by Identifying Spermatozoa RNA Isolation Methods and PRM1, TNP1, and MARCH 1 Gene Expression

Bali cattle are one of the local cattle breeds that have the highest population compared to other local cattle in Indonesia. Due to its superior resistance to parasites and tropical environments, it also has better reproductive efficiency. So that the development and preservation of Bali cattle from all aspects need to be carried out in a sustainable manner to support the effort supply of meat in Indonesia.

One way is to improve reproductive quality which is the main aspect in producing quality offspring, supported by an artificial insemination (AI) program. Basically the success of AI is influenced by superior male semen quality factors. To understand the molecular basis governing the productivity of semen, it is necessary to have groups of males with high and low productivity, so that the target genes expressed in the cells will be obtained.

This was revealed by a student of the doctoral program in animal science, Faculty of Animal Science, Universitas Brawijaya (Fapet UB), Chairdin Dwi Nugraha, S.Pt., M.Pt. in his research entitled“Development of RNA Isolation Methods and Analysis of PRM1, TNP1, and MARCH1 Gene Expression as Potential Marker Genes Basic for Selection of Prospective Male Superior Bali Cattle.”Research conducted with Prof. Suyadi (Fapet UB), Prof. Kuswati (Fapet UB), and Prof. Widodo (FMIPA). has been presented through an open dissertation exam, Monday (7/8/2023).

According to him, the application of gene expression technology can be utilized in Bali cattle in the analysis of genes related to semen productivity. On the other hand, sperm contains a very small amount of RNA, so it is necessary to develop the correct RNA isolation method. Extraction of RNA from sperm includes purification of the sperm and evaluation of RNA quality.

The study was conducted on 28 male Bali cattle through three stages. The first is the calculation of the Total Motile Spermatozoa (TSM) value in one year, the second is identifying the right method of isolating spermatozoa RNA with lysis modification, and the last step is studying the expression of genes derived from frozen semen for each individual in the high semen productivity group and the semen productivity group. low.

The results showed that male Bali cattle in the high productivity group had the potential to produce frozen semen up to 12,000 straws per year. The best sperm isolation uses the Direct-zol RNA Miniprep Kit with modifications, namely the incubation temperature of 80°C + crushing using a micropestle. Semen production in superior bulls of Bali cattle can be distinguished by the expression of the TNP1 and MARCH1 genes with a relative expression level of > 1.0. The role of the TNP1 and MARCH1 genes can potentially be used as candidate genes for semen productivity. (dta)