Whole genome RNA sequencing of spermatozoa from holstein bulls with various cryoresistance

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Whole-genome RNA sequencing (RNA-Seq) was performed to compare sperm transcript profiles from bulls with different sperm productivity characteristics and cryotolerance. Ejaculates from Holstein bulls classified as having high and low cryopreservability and sperm productivity (НСS and LCS), respectively; n = 3 bulls per group) were analyzed for post-thaw motility, mitochondrial membrane potential, plasma membrane integrity, and acrosomes. Total RNA was isolated from decryopreserved spermatozoa from GSF and PSF bulls and subjected to RNA-seq (Illumina NextSeq 500 platform), and transcript arrays were assessed using DESeq2, DESeq packages in each bull's spermatozoa. Several differentially expressed genes (DEGs) related to inflammation (IRG1, MMP12), spermatogenesis (Col4A2,), and unknown function (LOC 781178, LOC618942) were identified that were preferentially downregulated in ejaculates with poor cryotolerance.

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Sobre autores

O. Barkova

L.K. Ernst Federal Research Center for Animal Husbandry

Email: barkoffws@list.ru

All-Russian Research Institute of Genetics and Breeding of Farm Animals (VNIIGRZh)

Rússia, Pushkin, St. Petersburg

D. Starikova

L.K. Ernst Federal Research Center for Animal Husbandry

Email: barkoffws@list.ru

All-Russian Research Institute of Genetics and Breeding of Farm Animals (VNIIGRZh)

Rússia, Pushkin, St. Petersburg

I. Chistyakova

L.K. Ernst Federal Research Center for Animal Husbandry

Email: barkoffws@list.ru

All-Russian Research Institute of Genetics and Breeding of Farm Animals (VNIIGRZh)

Rússia, Pushkin, St. Petersburg

N. Pleshanov

L.K. Ernst Federal Research Center for Animal Husbandry

Autor responsável pela correspondência
Email: barkoffws@list.ru

All-Russian Research Institute of Genetics and Breeding of Farm Animals (VNIIGRZh)

Rússia, Pushkin, St. Petersburg

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2. Fig. 1. Productive traits of decryopreserved bull sperm. Values ​​are expressed as the mean (±SEM) of three ejaculates from each bull with high (black bar) and low (gray bar) sperm productivity indices (SPI and LPI, respectively). All indices had significant differences in sperm traits of at least P < 0.05.

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3. Fig. 2. Graphical representation of the results of principal component analysis (PCA) and multidimensional scaling (MDS) to determine the distance between sperm transcript samples. (a) PCA plot shows the distance from sample to sample for transcripts of six bulls from two groups (case – bulls from the NCS group, control – bulls from the VCS group); (b) MDS plot shows the distances between samples among bull replications. Blue dots indicate bulls with poor sperm cryoresistance (IRG1, MMP12, Col4A2, LOC 781178, LOC618942).

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4. Fig. 3. Volcano plot of the differentially expressed gene (DegS) in bull sperm. Each dot represents a single gene. Gray dots indicate genes whose expression was slightly differentially upregulated; blue dots indicate genes whose expression was significantly downregulated (log2FC > 1 or < –1, and adjusted P-value < 0.05; FDR < 0.05).

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5. Fig. 4. Gene ontology (GO) analysis of differentially expressed genes (DEGs) in bull spermatozoa differing in cryoresistant ability and fertility qualities.

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