Abstract
BackgroundCryptorchidism is a condition that occurs when one or both testes fail to descend into the scrotum. It is a common congenital disorder, causing economic loss in pig production. However, there have been only limited studies of differential protein expression profiles in undescended testes (UDTs) in the abdomen and descended testes (DTs) in cryptorchid pigs, especially at the peptidome and proteome levels. The present study aimed to analyze the peptidome of UDTs and DTs in unilateral cryptorchid pigs aged 1–2, 6, 15 and 20 weeks and in normal testes of healthy pigs aged 1–2 and 12 weeks, using peptide mass fingerprinting and three-dimensional principal component analysis (3D-PCA) with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and to identify potential protein candidates, using in-gel digestion coupled with mass spectrometry (GeLC-MS/MS). Western blot analysis was used to verify protein expression. Protein sequence was affirmed by liquid chromatography–tandem mass spectrometry.ResultsA PCA plot showed a discrete cluster for each sample group. Peptide mass fingerprints (PMFs) demonstrated unique peptide fragments in UDTs at different ages. A number of markedly expressed proteins from GeLC-MS/MS were identified, including the multifunctional tumor necrosis factor receptor superfamily member 18 (TNFRSF18), in DTs at 1–2 and 6 weeks and in UDTs at 15 and 20 weeks of age. Using western blot analysis, high expression of TNFRSF18 was observed in the UDTs at 15 weeks. Using the STITCH database, this protein was found to be related to apoptosis, corresponding to the previous report in the UDTs at the same age.ConclusionsThe present study revealed the specific PMFs and clusters for porcine cryptorchidism, and a novel protein, TNFRSF18, associated with the disease mechanism. These results could provide further insights into the pathogenesis of the disease.
Highlights
Cryptorchidism is a condition that occurs when one or both testes fail to descend into the scrotum
In-gel digestion coupled with mass spectrometry (GeLCMS/Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS)), a one-dimensional sodium dodecyl sulfate– polyacrylamide gel electrophoresis (SDS-PAGE) coupled with liquid chromatography–tandem mass spectrometry (LC-MS/MS), has been used for identification of proteins [11]
Distinct peptide peak spectra were observed in undescended testes in the abdominal cavity (UDT) at different ages and in Normal testis (NT) at 1–2 weeks of age (Fig. 1)
Summary
Cryptorchidism is a condition that occurs when one or both testes fail to descend into the scrotum It is a common congenital disorder, causing economic loss in pig production. There have been only limited studies of differential protein expression profiles in undescended testes (UDTs) in the abdomen and descended testes (DTs) in cryptorchid pigs, especially at the peptidome and proteome levels. Cryptorchidism is a condition that occurs when one or both testes fail to descend into the scrotum, which can reduce fertility and increase the risk of testicular malignancies [1]. The expression of protein markers could possibly be utilized for unveiling the aberrant mechanism underlying the disease, and for selection of suitable sires from the GGPs and GPs, which might reduce the incidence of cryptorchidism in fattening pigs. We decided to explore a panel of novel peptide and protein expression, using proteomic approaches
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