Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis
Accurate determination of estrus is essentially required for efficient reproduction management of farm animals. Buffalo is a shy breeder and does not manifest overt signs of estrus that make estrus detection difficult resulting in a poor conception rate. Therefore, identifying estrus biomarkers in e...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-05-01
|
| Series: | Frontiers in Genetics |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fgene.2022.867909/full |
| _version_ | 1828339400523644928 |
|---|---|
| author | Laishram Kipjen Singh Laishram Kipjen Singh Mamta Pandey Rubina Kumari Baithalu Rubina Kumari Baithalu Abhijeet Fernandes Syed Azmal Ali Syed Azmal Ali Latika Jaiswal Suryaprakash Pannu Neeraj Tushar K. Mohanty A. Kumaresan Tirtha K. Datta Sudarshan Kumar Ashok K. Mohanty |
| author_facet | Laishram Kipjen Singh Laishram Kipjen Singh Mamta Pandey Rubina Kumari Baithalu Rubina Kumari Baithalu Abhijeet Fernandes Syed Azmal Ali Syed Azmal Ali Latika Jaiswal Suryaprakash Pannu Neeraj Tushar K. Mohanty A. Kumaresan Tirtha K. Datta Sudarshan Kumar Ashok K. Mohanty |
| author_sort | Laishram Kipjen Singh |
| collection | DOAJ |
| description | Accurate determination of estrus is essentially required for efficient reproduction management of farm animals. Buffalo is a shy breeder and does not manifest overt signs of estrus that make estrus detection difficult resulting in a poor conception rate. Therefore, identifying estrus biomarkers in easily accessible biofluid such as saliva is of utmost interest. In the current study, we generated saliva proteome profiles during proestrus (PE), estrus (E), metestrus (ME), and diestrus (DE) stages of the buffalo estrous cycle using both label-free quantitation (LFQ) and labeled (TMT) quantitation and mass spectrometry analysis. A total of 520 proteins were identified as DEPs in LFQ; among these, 59 and four proteins were upregulated (FC ≥ 1.5) and downregulated (FC ≤ 0.5) during E vs. PE, ME, and DE comparisons, respectively. Similarly, TMT-LC-MS/MS analysis identified 369 DEPs; among these, 74 and 73 proteins were upregulated and downregulated during E vs. PE, ME, and DE stages, respectively. Functional annotations of GO terms showed enrichment of glycolysis, pyruvate metabolism, endopeptidase inhibitor activity, salivary secretion, innate immune response, calcium ion binding, oocyte meiosis, and estrogen signaling. Over-expression of SERPINB1, HSPA1A, VMO1, SDF4, LCN1, OBP, and ENO3 proteins during estrus was further confirmed by Western blotting. This is the first comprehensive report on differential proteome analysis of buffalo saliva between estrus and non-estrus stages. This study generated an important panel of candidate proteins that may be considered buffalo estrus biomarkers which can be applied in the development of a diagnostic kit for estrus detection in buffalo. |
| first_indexed | 2024-04-13T22:42:15Z |
| format | Article |
| id | doaj.art-d55e4986b54648eaac4947576ed736ab |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-04-13T22:42:15Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-d55e4986b54648eaac4947576ed736ab2022-12-22T02:26:35ZengFrontiers Media S.A.Frontiers in Genetics1664-80212022-05-011310.3389/fgene.2022.867909867909Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalisLaishram Kipjen Singh0Laishram Kipjen Singh1Mamta Pandey2Rubina Kumari Baithalu3Rubina Kumari Baithalu4Abhijeet Fernandes5Syed Azmal Ali6Syed Azmal Ali7Latika Jaiswal8Suryaprakash Pannu9 Neeraj10Tushar K. Mohanty11A. Kumaresan12Tirtha K. Datta13Sudarshan Kumar14Ashok K. Mohanty15Animal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaMolecular Reproduction Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaMolecular Reproduction Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaMolecular Reproduction Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaProteomics and Cell Biology Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaDivision of Proteomics of Stem Cells and Cancer, German Cancer Research Center (DKFZ), Heidelberg, GermanyMolecular Reproduction Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Reproduction, Gynaecology and Obstetrics, ICAR-National Dairy Research Institute, Karnal, IndiaAnimal Biotechnology Centre, ICAR-National Dairy Research Institute, Karnal, IndiaProteomics and Cell Biology Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaProteomics and Cell Biology Lab, Animal Biotechnology Center, ICAR-National Dairy Research Institute, Karnal, IndiaAccurate determination of estrus is essentially required for efficient reproduction management of farm animals. Buffalo is a shy breeder and does not manifest overt signs of estrus that make estrus detection difficult resulting in a poor conception rate. Therefore, identifying estrus biomarkers in easily accessible biofluid such as saliva is of utmost interest. In the current study, we generated saliva proteome profiles during proestrus (PE), estrus (E), metestrus (ME), and diestrus (DE) stages of the buffalo estrous cycle using both label-free quantitation (LFQ) and labeled (TMT) quantitation and mass spectrometry analysis. A total of 520 proteins were identified as DEPs in LFQ; among these, 59 and four proteins were upregulated (FC ≥ 1.5) and downregulated (FC ≤ 0.5) during E vs. PE, ME, and DE comparisons, respectively. Similarly, TMT-LC-MS/MS analysis identified 369 DEPs; among these, 74 and 73 proteins were upregulated and downregulated during E vs. PE, ME, and DE stages, respectively. Functional annotations of GO terms showed enrichment of glycolysis, pyruvate metabolism, endopeptidase inhibitor activity, salivary secretion, innate immune response, calcium ion binding, oocyte meiosis, and estrogen signaling. Over-expression of SERPINB1, HSPA1A, VMO1, SDF4, LCN1, OBP, and ENO3 proteins during estrus was further confirmed by Western blotting. This is the first comprehensive report on differential proteome analysis of buffalo saliva between estrus and non-estrus stages. This study generated an important panel of candidate proteins that may be considered buffalo estrus biomarkers which can be applied in the development of a diagnostic kit for estrus detection in buffalo.https://www.frontiersin.org/articles/10.3389/fgene.2022.867909/fullbiomarkerestruslabel-free quantitationLC-MS/MStandem mass tagsaliva |
| spellingShingle | Laishram Kipjen Singh Laishram Kipjen Singh Mamta Pandey Rubina Kumari Baithalu Rubina Kumari Baithalu Abhijeet Fernandes Syed Azmal Ali Syed Azmal Ali Latika Jaiswal Suryaprakash Pannu Neeraj Tushar K. Mohanty A. Kumaresan Tirtha K. Datta Sudarshan Kumar Ashok K. Mohanty Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis Frontiers in Genetics biomarker estrus label-free quantitation LC-MS/MS tandem mass tag saliva |
| title | Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis |
| title_full | Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis |
| title_fullStr | Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis |
| title_full_unstemmed | Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis |
| title_short | Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis |
| title_sort | comparative proteome profiling of saliva between estrus and non estrus stages by employing label free quantitation lfq and tandem mass tag tmt lc ms ms analysis an approach for estrus biomarker identification in bubalus bubalis |
| topic | biomarker estrus label-free quantitation LC-MS/MS tandem mass tag saliva |
| url | https://www.frontiersin.org/articles/10.3389/fgene.2022.867909/full |
| work_keys_str_mv | AT laishramkipjensingh comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT laishramkipjensingh comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT mamtapandey comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT rubinakumaribaithalu comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT rubinakumaribaithalu comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT abhijeetfernandes comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT syedazmalali comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT syedazmalali comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT latikajaiswal comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT suryaprakashpannu comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT neeraj comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT tusharkmohanty comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT akumaresan comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT tirthakdatta comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT sudarshankumar comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis AT ashokkmohanty comparativeproteomeprofilingofsalivabetweenestrusandnonestrusstagesbyemployinglabelfreequantitationlfqandtandemmasstagtmtlcmsmsanalysisanapproachforestrusbiomarkeridentificationinbubalusbubalis |