Cryopreservation‐induced alterations in protein composition of rainbow trout semen

Abstract

The aim of this study was to detect cryopreservation-induced alterations in the protein composition of rainbow trout semen using two independent methods 1DE SDS-PAGE prefractionation combined with LC-MS/MS and 2D difference gel electrophoresis followed by MALDI-TOF/TOF identification. Here, we show the first comprehensive dataset of changes in rainbow trout semen proteome after cryopreservation, with a total of 73 identified proteins released from sperm to extracellular fluid, including mitochondrial, cytoskeletal, nuclear, and cytosolic proteins. Our study provides new information about proteins released from sperm, their relation to sperm structure and function, and changes of metabolism of sperm cells as a result of cryopreservation. The identified proteins represent potential markers of cryoinjures of sperm structures and markers of the disturbances of particular sperm metabolic pathways. Further studies will allow to decipher the precise function of the proteins altered during rainbow trout cryopreservation and are useful for the development of extensive diagnostic tests of sperm cryoinjures and for the successful improvement of sperm cryopreservation of this economically important species.

Transcriptome Sequencing Reveals the Differentially Expressed lncRNAs and mRNAs Involved in Cryoinjuries in Frozen-Thawed Giant Panda (Ailuropoda melanoleuca) Sperm

“…The mRNAs targeted by these lncRNAs and the differentially expressed mRNA were mainly enriched in membrane-related terms (integral component of membrane and membrane) and responses to stimulus. Similar GO enrichment results were observed in GO analysis of differentially expressed proteins in rainbow trout frozen sperm [ 44 ] and differentially expressed miRNAs of porcine frozen sperm [ 18 ]. Moreover, these membrane-related terms could be associated with sperm cryodamage.…”

Proteomic identification of rainbow trout blood plasma proteins and their relationship to seminal plasma proteins

“…Spots of interests were manually excised, subjected to tryptic digestion and identified as described by Nynca et al. by an Autoflex TOF/TOF mass spectrometer (MALDI‐TOF/TOF; Bruker Daltonics, Bremen, Germany). MS peptide mass fingerprint and fragment spectra from each individual spot were combined and used to search against the NCBInr fish database (released on April 10, 2014) using the MASCOT search engine (version 2.4, Matrix Science Ltd., London, UK) in consideration of the following settings: (i) enzyme trypsin; (ii) fixed modifications carbamidomethyl; (iii) variable modifications methionine oxidation; (iv) allow to miss one cleavage; (v) peptide tol.…”

“…Corresponding proteins of SP and BP were considered to be more abundant in SP if they fulfilled the following enrichment criteria: (i) normalised spectral counts need to be at least 2.5 times higher in SP compared to the corresponding BP data and (ii) at least ten spectra per protein in at least one sample need to be acquired. A similar strategy has been applied by Flenkenthaler et al [15] and Nynca et al [16]. Statistical analysis of protein abundance was also performed (Section 2.8).…”

Cryomedia Formula: Cellular Molecular Perspective

“…Furthermore, the cryopreservation affects cells' biomarkers [56]. It alters the proteome profile of cells, which in some cases can bring about changes in cellular metabolism, function, and structure [57]. In previous work, there is often no clear demarcation between the effect of CPAs and the cryopreservation protocol itself.…”