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Title:	Comparative proteomics and metabolomics of sperm from Kedah Kelantan (Bos indicus) and mafriwal (Bos taurus x Bos inducus) cattle
Authors:	Ashrafzadeh Ali (P48920)
Supervisor:	Sheila Nathan, Prof. Dr.
Keywords:	Fertility of Zebu cattle Metabolomics- Proteomics Dissertations, Academic -- Malaysia
Issue Date:	17-Jun-2015
Description:	In tropical areas, the fertility of Zebu cattle (Bos indicus) is higher than that of the European purebred (Bos taurus) and crossbreed (Bos taurus × Bos indicus). To identify proteins, metabolites and metabolic pathways attributed to the higher fertility of Zebu cattle (B. indicus) sperm in tropical areas, integration of metabolomics- and proteomics-based approaches was used to unravel physiological differences between sperm from the high fertile Malaysian indigenous breed, Kedah Kelantan (KK, B. indicus), and sperm from the sub-fertile crossbreed of cattle, Mafriwal (M, B. taurus × B. indicus). Two-dimensional polyacrylamide gel electrophoresis and the integration of shotgun metabolomics and proteomics using liquid chromatography mass spectrometry successfully compared sperm proteins and metabolites between KK and M. A number of proteins and metabolites associated with glycolysis (e.g α-D-glucose and glyceraldehyde-3-phosphate dehydrogenase), tricarboxylic acid cycle (e.g. isocitrate and isocitrate dehydrogenase), electron-transport chain (e.g. ubiquinol and NADH dehydrogenase), lipid metabolism (e.g. fatty acids and carnitine Oacyltransferase), response to stress (e.g. heat shock protein family members), spermoocyte interaction (e.g. zona pellucida binding protein) and flagella structure (e.g. tubulin family members) that are crucial in sperm fertility were shown to be regulated in KK and M. The results suggest more efficient carbohydrate and energy metabolism, specifically through glycolysis, in KK. On the other hand, lipid metabolism relatedproteins which were specific to M suggest a physiological compensation associated with defects in carbohydrate metabolism or a breed-specific characteristic which requires further validation. To confirm the proteomics and metabolomics findings, sperm motility, sperm morphology and plasma membrane integrity, DNA fragmentation and the level of reactive oxygen species were evaluated using different procedures. The increased sperm motility observed was consistent with the higher abundance of twelve flagella structural proteins in KK. Lower DNA fragmentation and higher plasma membrane integrity as well as lower levels of reactive oxygen species were apparent in KK. The higher sperm plasma membrane integrity in KK is most likely related to the higher level of phospholipids observed. A more robust antioxidant system and reduced polyunsaturated fatty acids content may help KK to cope with oxidative stress under the warm conditions of the tropics. As proteins involved in sperm-oocyte interaction are up-regulated in sperm from M, the lower fertility rates of M cattle may not be directly attributed to possible defects in these proteins. The findings from this study propose that the robust antioxidant activity, higher DNA and plasma membrane integrity, stronger flagella structure and energy production through glycolysis and higher motility of KK sperm contribute to the higher fertility of this breed in the tropics.,Di kawasan tropika, kesuburan lembu Zebu (Bos indicus) adalah lebih tinggi berbanding baka tulen Eropah (Bos taurus) dan baka kacukan (Bos taurus × Bos indicus). Bagi mengenalpasti protein, metabolit dan tapak jalan metabolik yang berperanan dalam kesuburan sperma lembu Zebu, pendekatan bersepadu yang melibatkan kaedah metabolomik dan proteomik telah digunakan. Ini dilakukan untuk merungkai perbezaan fisiologi di antara sperma daripada baka asli Malaysia yang berkesuburan tinggi, Kedah Kelantan (KK, B. indicus), dan sperma daripada baka kacukan berkesuburan rendah, Mafriwal (M, B. taurus × B. indicus). Elektroforesis gel dua dimensi dan gabungan kaedah metabolomik dan proteomik "shotgun" menggunakan kromatografi cecair spektrometri jisim telah berjaya membandingkan protein dan metabolit sperma di antara KK dan M. Beberapa protein dan metabolit yang terlibat dalam glikolisis (cth: α-D-glukosa dan gliseraldehid 3-fosfat dehydrogenase), kitaran asid trikarboksilik (cth: isositrat dan isositrat dehydrogenase) , rantaian pengangkutan electron (cth: ubiquinol dan NADH dehydrogenase), metabolisma lipid (cth: asid lemak dan karnitina O-asiltransferase), gerak balas terhadap tekanan (cth: protein kejutan haba), interaksi sperma-oosit (cth: zona pellucida protein mengikat) dan struktur flagella (cth: tubulin) didapati berbeza ekspresinya dalam KK dan M. Keputusan ini mencadangkan metabolisme karbohidrat dan tenaga yang lebih cekap dalam KK, khususnya yang melibatkan tapak jalan glikolisis. Sebaliknya, protein yang berkait dengan metabolisme lipid didapati hadir hanya dalam M dan ini mencadangkan pampasan fisiologi yang berkaitan dengan kecacatan metabolisme karbohidrat atau berkait-baka. Walaubagaimanapun, pemerhatian ini perlu ditentusahkan. Untuk mengesahkan keputusan analisis proteom dan metabolom, motiliti sperma, morfologi sperma dan integriti membran plasma, fragmentasi DNA dan tahap spesies oksigen reaktif telah dinilai menggunakan beberapa kaedah yang berlainan. Motiliti sperma KK yang lebih tinggi adalah konsisten dengan jumlah protein struktur flagela yang lebih tinggi dalam KK. Tahap fragmentasi DNA yang rendah dan integriti membran plasma yang lebih tinggi serta tahap spesies oksigen reaktif yang lebih rendah diperhatikan dalam KK. Integriti membran plasma sperma yang lebih tinggi dalam KK besar kemungkinannya berkait dengan kandungan fosfolipid yang lebih tinggi. Hasil penemuan kajian ini memandangkan bahawa aktiviti antioksidan yang teguh, integriti DNA dan membran plasma yang lebih tinggi, struktur flagella yang lebih kukuh dan penghasilan tenaga melalui glikolisis serta motiliti sperma yang lebih tinggi dalam KK menyumbang kepada kesuburan yang lebih tinggi baka ini di kawasan tropika.,Ph.D
Pages:	163
Call Number:	QP551 .A839 2015
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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