Studies on substantially increased proteins in follicular fluid of bovine ovarian follicular cysts using 2-D PAGE and MA
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Studies on substantially increased proteins in follicular fluid of bovine ovarian follicular cysts using 2-D PAGE and MALDI-TOF MS Jiro Maniwa*1,2, Shunsuke Izumi3, Naoki Isobe1 and Takato Terada1 Address: 1Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan, 2Preclinical Sciences Department, AstraZeneca KK, Osaka 531-0076, Japan and 3Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8526, Japan Email: Jiro Maniwa* - [email protected]; Shunsuke Izumi - [email protected]; Naoki Isobe - [email protected]; Takato Terada - [email protected] * Corresponding author
Published: 08 June 2005 Reproductive Biology and Endocrinology 2005, 3:23 23
doi:10.1186/1477-7827-3-
Received: 02 May 2005 Accepted: 08 June 2005
This article is available from: http://www.rbej.com/content/3/1/23 © 2005 Maniwa et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: The objective of this study was to identify substantially increased proteins in bovine cystic follicular fluid (FF) in order to clarify the pathology and etiology of bovine ovarian follicular cysts (BOFC). Methods: Proteins in normal and cystic FF samples were subjected to two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and were compared using silver stained gel images with PDQuest image analysis software. Peptides from these increased spots were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), and were identified based on the NCBI database by a peptide mass fingerprinting method. Results: Comparative proteomic analysis showed 8 increased protein spots present in cystic FF. MS analysis and database searching revealed that the increased proteins in cystic FF were bovine mitochondrial f1-atpase (BMFA), erythroid associated factor (EAF), methionine synthase (MeS), VEGF-receptor, glyceraldehydes 3-phosphate dehydrogenase (GAPDH), heat shock protein 70 (HSP70), β-lactoglobulin (BLG) and succinate dehydrogenase Ip subunit (SD). Conclusion: Our results suggest that these proteins are overexpressed in BOFC, and that they may play important roles in the pathogenesis of BOFC. Furthermore, these proteins in the FF could be useful biomarkers for BOFC.
Background BOFC is one of the most frequently diagnosed gynecological findings in cattle, and is a major reproductive problem in cows, causing infertility [1-3]. BOFC is generally defined as follicle-like structures of greater than 25 mm in diameter without a corpus luteum in both ovaries [1,4]. There have been several different hypotheses regarding the cause of BOFC, such as inherited factors, high lacta-
tion, aging, seasonal effects, nutrition
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