Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS
In sperm proteomic experiments round cells and leukocyte proteins are profiled along with sperm proteome. The influence of round cell and leukocyte proteins on the sperm proteome has not been investigated. The objective of this study was to identify if the proteins from round cells, including leukoc...
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doaj-b5edab829eb547a49cfdd9832d83bcd22020-11-25T00:30:44ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-01-0120231410.3390/ijms20020314ijms20020314Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MSManesh Kumar Panner Selvam0Ashok Agarwal1Tânia R. Dias2Ana D. Martins3Luna Samanta4American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USAAmerican Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USAAmerican Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USAAmerican Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USAAmerican Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USAIn sperm proteomic experiments round cells and leukocyte proteins are profiled along with sperm proteome. The influence of round cell and leukocyte proteins on the sperm proteome has not been investigated. The objective of this study was to identify if the proteins from round cells, including leukocytes, interfere with the proteomic analysis of spermatozoa in frozen semen samples. Proteomic profiling of sperm was performed using liquid chromatography-tandem mass spectrometry in four groups: Group 1 contained neat semen with round cells and leukocytes ≥ 1 × 106/mL, group 2 contained neat semen with round cells ≥ 1 × 106/mL that was processed by 65% density gradient to remove the round cells and leukocytes, group 3 contained neat semen with round cells < 1 × 106/mL, and group 4 contained neat semen with round cells < 1 × 106/mL that was processed by 65% density gradient to remove the round cells. Pure leukocyte culture was used as control group. A total of 1638, 1393, 1755, and 1404 proteins were identified in groups 1, 2, 3, and 4, respectively. Comparative analysis of group 1 vs. 3 revealed 26 (1.18%) differentially expressed proteins (DEPs). On the other hand, only 6 (0.31%) DEPs were observed with group 2 vs. 4. Expression of these DEPs were either absent or very low in the control group. The results of our proteomics analysis failed to show any influence of non-spermatogenic round cell proteins on sperm proteome identification. These results validate the use of neat semen samples for sperm proteomic studies.http://www.mdpi.com/1422-0067/20/2/314spermleukocytesround cellsnon-spermatogenic cellsfrozen semenproteomics |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Manesh Kumar Panner Selvam Ashok Agarwal Tânia R. Dias Ana D. Martins Luna Samanta |
spellingShingle |
Manesh Kumar Panner Selvam Ashok Agarwal Tânia R. Dias Ana D. Martins Luna Samanta Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS International Journal of Molecular Sciences sperm leukocytes round cells non-spermatogenic cells frozen semen proteomics |
author_facet |
Manesh Kumar Panner Selvam Ashok Agarwal Tânia R. Dias Ana D. Martins Luna Samanta |
author_sort |
Manesh Kumar Panner Selvam |
title |
Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS |
title_short |
Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS |
title_full |
Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS |
title_fullStr |
Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS |
title_full_unstemmed |
Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS |
title_sort |
presence of round cells proteins do not interfere with identification of human sperm proteins from frozen semen samples by lc-ms/ms |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2019-01-01 |
description |
In sperm proteomic experiments round cells and leukocyte proteins are profiled along with sperm proteome. The influence of round cell and leukocyte proteins on the sperm proteome has not been investigated. The objective of this study was to identify if the proteins from round cells, including leukocytes, interfere with the proteomic analysis of spermatozoa in frozen semen samples. Proteomic profiling of sperm was performed using liquid chromatography-tandem mass spectrometry in four groups: Group 1 contained neat semen with round cells and leukocytes ≥ 1 × 106/mL, group 2 contained neat semen with round cells ≥ 1 × 106/mL that was processed by 65% density gradient to remove the round cells and leukocytes, group 3 contained neat semen with round cells < 1 × 106/mL, and group 4 contained neat semen with round cells < 1 × 106/mL that was processed by 65% density gradient to remove the round cells. Pure leukocyte culture was used as control group. A total of 1638, 1393, 1755, and 1404 proteins were identified in groups 1, 2, 3, and 4, respectively. Comparative analysis of group 1 vs. 3 revealed 26 (1.18%) differentially expressed proteins (DEPs). On the other hand, only 6 (0.31%) DEPs were observed with group 2 vs. 4. Expression of these DEPs were either absent or very low in the control group. The results of our proteomics analysis failed to show any influence of non-spermatogenic round cell proteins on sperm proteome identification. These results validate the use of neat semen samples for sperm proteomic studies. |
topic |
sperm leukocytes round cells non-spermatogenic cells frozen semen proteomics |
url |
http://www.mdpi.com/1422-0067/20/2/314 |
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