Quantitative proteomic profiling of bovine follicular fluid during follicle development

Author(s)

Ferrazza, Rodrigo de Andrade ^[1] ; Mogollon Garcia, Henry David ^[1] ; dos Santos Schmidt, Elizabeth Moreira ^[2] ; Carmichael, Monika Mihm ^[3] ; de Souza, Fabiana Ferreira ^[1] ; Burchmore, Richard ^[4] ; Sartori, Roberto ^[5] ; Eckersall, Peter David ^[3] ; Pinheiro Ferreira, Joao Carlos ^[1]

Affiliation

^[1] Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Anim Reprod & Vet Radiol, Botucatu, SP - Brazil

^[2] Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP - Brazil

^[3] Univ Glasgow, Sch Vet Med, Inst Biodivers Anim Hlth & Comparat Med, Glasgow, Lanark - Scotland

^[4] Univ Glasgow, Coll Med Vet & Life Sci, Glasgow Poly Facil, Glasgow, Lanark - Scotland

^[5] Univ Sao Paulo, Luiz de Queiroz Coll Agr, Dept Anim Sci, Sao Paulo - Brazil

• Document type: Journal article
• Source: BIOLOGY OF REPRODUCTION; v. 97, n. 6, p. 835-849, DEC 2017.
• Web of Science Citations: 2

Abstract

Bovine follicular fluid (FF) constitutes the microenvironment of follicles and includes various biologically active proteins. We performed a study involving 18 healthy nonlactating Holstein cows to determine the protein expression profile of FF at key stages of follicular development. Follicles were individually aspirated in vivo at predeviation (F1 similar to 7.0 mm), deviation (F1 similar to 8.5 mm), postdeviation (F1 similar to 12.0 mm), and preovulatory stages of follicle development, which were confirmed by measurement of follicular estradiol and progesterone concentrations. The FFs from nine cows were selected for proteomic analysis. After albumin depletion, triplicates of pooled FF were reduced, alkylated, and digested with trypsin. The resulting peptides were labeled with TMTsixplex and quantified using liquid chromatography-mass spectrometry/mass spectrometry. A total of 143 proteins were identified and assigned to a variety of biological processes, including response to stimulus and metabolic processes. Twenty-two differentially (P < 0.05) expressed proteins were found between stages indicating intrafollicular changes over development, with expected deviation time critical to modulate the protein expression. For instance, high concentrations of follistatin, inhibin, serglycin, spondin-1, fibrinogen, and anti-testosterone antibody were found during early stages of follicular development. In contrast, apolipoprotein H, alpha-2-macroglobulin, plasminogen, antithrombin-III, and immunoglobulins were increased after deviation. Among the differentially abundant proteins, 19 were found to be associated with steroidogenesis. Pathway analysis identified proteins that were mainly associated with the acute phase response signaling, coagulation system, complement system, liver/retinoid X receptor activation, and biosynthesis of nitric oxide and reactive oxygen. The differentially expressed proteins provide insights into the size-dependent protein changes in the ovarian follicle microenvironment that could influence follicular function. Summary Sentence The protein dynamic changes in ovarian follicle microenvironment during the follicle development are critical for follicular maturation and influence follicular function in cows. (AU)

• FAPESP's process: 12/18297-7 - Effect of heat stress on follicular development and luteolysis in Honstein cows, role of LH during induced luteolysis and characterization of luteal sensitivity to PGF2± during the estrous cycle
• Grantee: João Carlos Pinheiro Ferreira
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/20083-8 - In vivo study of the factors involved in the establishment of follicular dominance and atresia in Holstein cows and the effect of heat stress on these phenomena
• Grantee: Rodrigo de Andrade Ferrazza
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 14/21257-2 - Proteomics evaluation of the follicular fluid of Holstein cows under heat stress
• Grantee: Rodrigo de Andrade Ferrazza
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate