Abstract

Spermatozoa carries a reservoir of mRNAs regulating sperm functions and fertilizing potential. Although it is well recognized that a considerable proportion of high genetic merit breeding bulls produce poor-quality semen, the transcriptomic alterations in spermatozoa from such bulls are not understood. In the present study, comparative high-throughput transcriptomic profiling of spermatozoa from good and poor-quality semen-producing bulls was carried out to identify the transcripts associated with semen quality. Using next-generation sequencing (NGS), we identified 11,632 transcripts in Holstein Friesian bull spermatozoa; after total hit normalization, a total of 544 transcripts were detected, of which 185 transcripts were common to both good and poor-quality semen, while 181 sperm transcripts were unique to good quality semen, and 178 transcripts were unique to poor-quality semen. Among the co-expressed transcripts, 31 were upregulated, while 108 were downregulated, and 46 were neutrally expressed in poor-quality semen. Bioinformatics analysis revealed that the dysregulated transcripts were predominantly involved in molecular function, such as olfactory receptor activity and odor binding, and in biological process, such as detection of chemical stimulus involved in sensory perception, sensory perception of smell, signal transduction, and signal synaptic transmission. Since a majority of the dysregulated transcripts were involved in the olfactory pathway (85% of enriched dysregulated genes were involved in this pathway), the expression of selected five transcripts associated with this pathway (OR2T11, OR10S1, ORIL3, OR5M11, and PRRX1) were validated using real-time qPCR, and it was found that their transcriptional abundance followed the same trend as observed in NGS; the sperm transcriptional abundance of OR2T11 and OR10S1 differed significantly (p < 0.05) between good and poor-quality semen. It is concluded that poor-quality semen showed altered expression of transcripts associated with olfactory receptors and pathways indicating the relationship between olfactory pathway and semen quality in bulls.

Highlights

  • For genetic improvement and enhancing productivity, artificial insemination (AI) of cows with high-genetic merit bulls is being carried out

  • Using Illumina Seq-500 RNA sequencing platforms, we identified 15,028,305 and 15,373,636 with paired-end 76 bp raw reads in spermatozoa of good and poor-quality semen, respectively

  • The processed reads were mapped to the reference genome (Bos taurus), which showed an alignment of ≃73% and ≃83% for good and poor-quality semen, respectively

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Summary

Introduction

For genetic improvement and enhancing productivity, artificial insemination (AI) of cows with high-genetic merit bulls is being carried out. AI of native cows with cryopreserved spermatozoa from high genetic exotic bulls/crossbred bulls improved the milk production substantially, the coverage of AI is limited in these countries. A significant proportion of bulls qualified through the BSE are culled because of poor-quality semen and subfertility during later stages. 20–25% variation in conception rates was observed among the bulls that qualified through the BSE [3, 4], and a substantial proportion of bulls that have passed the BSE had high ejaculation rejection rates [5, 6]. It has been reported that ejaculate rejection rate in breeding bulls ranged from 10 to 100% with the average being 52 to 55% [7–9]. High ejaculate rejection rates lead to colossal loss to the breeders and hamper genetic improvement

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