Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation
This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine ( Canis lupus familiaris ) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation...
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ftdoajarticles:oai:doaj.org/article:331d93b870bf4c87959e4a5a3097eee3 2023-05-15T15:49:48+02:00 Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation Anna Zmudzinska Mariusz A. Bromke Rafal Strzezek Magdalena Zielinska Beata Olejnik Marzena Mogielnicka-Brzozowska 2022-03-01T00:00:00Z https://doi.org/10.3390/ani12060772 https://doaj.org/article/331d93b870bf4c87959e4a5a3097eee3 EN eng MDPI AG https://www.mdpi.com/2076-2615/12/6/772 https://doaj.org/toc/2076-2615 doi:10.3390/ani12060772 2076-2615 https://doaj.org/article/331d93b870bf4c87959e4a5a3097eee3 Animals, Vol 12, Iss 772, p 772 (2022) epididymal spermatozoa semen quality proteomic sonication canine Veterinary medicine SF600-1100 Zoology QL1-991 article 2022 ftdoajarticles https://doi.org/10.3390/ani12060772 2022-12-31T04:10:52Z This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine ( Canis lupus familiaris ) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation. Sperm samples were collected from 15 dogs. Sperm protein fractions: intracellular (SIPs) and membrane-associated (SMAPs) were isolated. After sonication, sperm morphology was evaluated using Spermac Stain™. The sperm protein fractions were analyzed using gel electrophoresis (SDS-PAGE) and nanoliquid chromatography coupled to quadrupole time-of-flight mass spectrometry (NanoLC-Q-TOF/MS). UniProt database-supported identification resulted in 42 proteins identified in the SIPs and 153 proteins in the SMAPs. Differentially abundant proteins (DAPs) were found in SIPs and SMAPs. Based on a gene ontology analysis, the dominant molecular functions of SIPs were catalytic activity (50%) and binding (28%). Hydrolase activity (33%) and transferase activity (21%) functions were dominant for SMAPs. Bioinformatic analysis of SIPs and SMAPs showed their participation in important metabolic pathways in epididymal sperm, which may suggest their potential as sperm quality biomarkers. The use of sonication 150 W, 10 min, may be recommended for the separation of dog epididymal sperm heads, tails, acrosomes and the protein isolation. Article in Journal/Newspaper Canis lupus Directory of Open Access Journals: DOAJ Articles Animals 12 6 772 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
epididymal spermatozoa semen quality proteomic sonication canine Veterinary medicine SF600-1100 Zoology QL1-991 |
spellingShingle |
epididymal spermatozoa semen quality proteomic sonication canine Veterinary medicine SF600-1100 Zoology QL1-991 Anna Zmudzinska Mariusz A. Bromke Rafal Strzezek Magdalena Zielinska Beata Olejnik Marzena Mogielnicka-Brzozowska Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
topic_facet |
epididymal spermatozoa semen quality proteomic sonication canine Veterinary medicine SF600-1100 Zoology QL1-991 |
description |
This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine ( Canis lupus familiaris ) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation. Sperm samples were collected from 15 dogs. Sperm protein fractions: intracellular (SIPs) and membrane-associated (SMAPs) were isolated. After sonication, sperm morphology was evaluated using Spermac Stain™. The sperm protein fractions were analyzed using gel electrophoresis (SDS-PAGE) and nanoliquid chromatography coupled to quadrupole time-of-flight mass spectrometry (NanoLC-Q-TOF/MS). UniProt database-supported identification resulted in 42 proteins identified in the SIPs and 153 proteins in the SMAPs. Differentially abundant proteins (DAPs) were found in SIPs and SMAPs. Based on a gene ontology analysis, the dominant molecular functions of SIPs were catalytic activity (50%) and binding (28%). Hydrolase activity (33%) and transferase activity (21%) functions were dominant for SMAPs. Bioinformatic analysis of SIPs and SMAPs showed their participation in important metabolic pathways in epididymal sperm, which may suggest their potential as sperm quality biomarkers. The use of sonication 150 W, 10 min, may be recommended for the separation of dog epididymal sperm heads, tails, acrosomes and the protein isolation. |
format |
Article in Journal/Newspaper |
author |
Anna Zmudzinska Mariusz A. Bromke Rafal Strzezek Magdalena Zielinska Beata Olejnik Marzena Mogielnicka-Brzozowska |
author_facet |
Anna Zmudzinska Mariusz A. Bromke Rafal Strzezek Magdalena Zielinska Beata Olejnik Marzena Mogielnicka-Brzozowska |
author_sort |
Anna Zmudzinska |
title |
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
title_short |
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
title_full |
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
title_fullStr |
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
title_full_unstemmed |
Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine ( Canis lupus familiaris ) Epididymal Spermatozoa and Sperm Structure Separation |
title_sort |
proteomic analysis of intracellular and membrane-associated fractions of canine ( canis lupus familiaris ) epididymal spermatozoa and sperm structure separation |
publisher |
MDPI AG |
publishDate |
2022 |
url |
https://doi.org/10.3390/ani12060772 https://doaj.org/article/331d93b870bf4c87959e4a5a3097eee3 |
genre |
Canis lupus |
genre_facet |
Canis lupus |
op_source |
Animals, Vol 12, Iss 772, p 772 (2022) |
op_relation |
https://www.mdpi.com/2076-2615/12/6/772 https://doaj.org/toc/2076-2615 doi:10.3390/ani12060772 2076-2615 https://doaj.org/article/331d93b870bf4c87959e4a5a3097eee3 |
op_doi |
https://doi.org/10.3390/ani12060772 |
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Animals |
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12 |
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6 |
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772 |
_version_ |
1766384822956064768 |