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Evaluation of burbot Lota lota spermatozoon motility, lipid composition, protein profile, and swelling rate following change of temperature and osmolality

ABSTRACT: Despite available information on the roles of osmotic pressure, potassium (K+), and calcium (Ca2+) in activation of Eurasian burbot spermatozoon motility, the changes in sperm biochemical composition and mechanisms underlying their activation at temperatures above optimal spawning temperature is still unclear. We explored sperm swelling, lipid composition, and proteome in relation to osmolality and temperature of swimming medium. Higher temperatures increase burbot spermatozoa vulnerability to osmotic pressure, decreasing motility in extremely hypotonic media, and the presence of Ca2+ decreases swelling of spermatozoa. A role of swelling in activation of spermatozoa motility at high temperature was rejected. We observed no differences in protein profile or lipid composition with respect to activation of burbot spermatozoa at high temperature. Burbot sperm activation occurs at higher temperatures without modification of the spermatozoon membrane. Elucidation of the intrinsic signaling pathway of burbot spermatozoon spontaneous activation requires further study.

INSTRUMENT(S): autoflex

ORGANISM(S): Lota Lota

TISSUE(S): Spermatozoon

SUBMITTER: Dmitry Loginov

LAB HEAD: Jan Sterba

PROVIDER: PXD027402 | Pride | 2022-09-20

REPOSITORIES: Pride

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			Action	DRS
	191114_Hadiseh_samples_2D.tar	Other
	2D_results.csv	Csv
	Gadus_morhua_ncbi_191115.fasta	Fasta

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Project description:BACKGROUND: In previous studies using candidate gene approaches, low sperm count (oligospermia) has been associated with altered sperm mRNA content and DNA methylation in both imprinted and non-imprinted genes. We performed a genome-wide analysis of sperm DNA methylation and mRNA content to test for associations with sperm function. METHODS AND RESULTS: Sperm DNA and mRNA were isolated from 21 men with a range of semen parameters presenting to a tertiary male reproductive health clinic. DNA methylation was measured with the Illumina Infinium array at 27,000 CpG loci. Unsupervised clustering of methylation data differentiated the 21 sperm samples by their motility values. Recursively partitioned mixture modeling (RPMM) of methylation data resulted in four distinct methylation profiles that were significantly associated with sperm motility (P=0.01). Linear models of microarray analysis (LIMMA) was performed based on motility and identified 9,189 CpG loci with significantly altered methylation (Q<0.05) in the low motility samples, with many loci located in genes associated with subfertility and epigenetic regulation. In the low motility samples, the majority of disrupted CpG loci (80%) were hypomethylated. Of the aberrantly methylated CpGs, 194 were associated with imprinted genes almost equally distributed into hypermethylated (predominantly paternally expressed) and hypomethylated (predominantly maternally expressed) groups. Sperm mRNA was measured with the Human Gene 1.0 ST Affymetrix GeneChip Array. LIMMA analysis based on motility identified 20 candidate transcripts as differentially expressed in low motility sperm, including HDAC1 (NCBI 3065), SIRT3 (NCBI 23410), and DNMT3A (NCBI 1788). Altered expression of these epigenetic regulatory genes was associated with RPMM DNA methylation class. CONCLUSIONS: Using integrative genome-wide approaches to study epigenetic and gene expression patterns in human sperm we identified CpG methylation profiles and mRNA alterations associated with low sperm motility, and that low motility sperm may have aberrant genome-wide hypomethylation due to excess HDAC1 activity. See "summary" above

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Evaluation of burbot Lota lota spermatozoon motility, lipid composition, protein profile, and swelling rate following change of temperature and osmolality

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