title: Anti-Mullerian hormone (AMH) concentration in follicular fluid and mRNA expression of AMH receptor type II and LH receptor in granulosa cells as predictive markers of good buffalo (Bubalus bubalis) donors.
abstract: High individual variability in follicular recruitment and hence in the number of embryos produced is a major factor limiting the application of reproductive technologies in buffalo. Therefore, the identification of reliable markers to select embryo donors is critical to enroll buffaloes in embryo production programs. Better understanding of factors involved in follicular growth is also necessary to improve the response to superovulation in this species. The aim of this work was thus to determine the anti-Mullerian hormone (AMH) concentration in follicular fluid (FF) recovered from different size follicles and evaluate the mRNA expression profiles of development-related (AMHR2, CYP19A1, FSHR, and LHR) and apoptosis-related genes (TP53INP1 and CASP3) in the corresponding granulosa cells (GCs) in buffalo. Another objective was to evaluate whether the AMH concentration in FF and gene expression of GCs is associated with the antral follicular count. Ovaries were collected at the slaughterhouse, and all follicles were counted and classified as small (3-5 mm), medium (5-8 mm), and large (>8 mm). Follicular fluid was recovered for AMH determination, and the mRNA expression of AMHR2, FSHR, LHR, CYP19A1, TP53INP1, and CASP3 was analyzed in GCs. The AMH concentration in FF decreased (P < 0.01) at increasing follicular diameter. The mRNA expression of AMHR2 and FSHR was higher (P < 0.05) in small follicles, whereas that of LHR and CYP19A1 was higher (P < 0.05) in large follicles. The intrafollicular AMH concentration was positively correlated with the antral follicular count (r = 0.31; P < 0.05). Interestingly, good donors (≥12 follicles) had a higher (P < 0.05) concentration of AMH and AMHR2 levels in small follicles and higher (P < 0.05) LHR levels in large follicles than bad donors (<12 follicles). These results suggest a potential use of AMH to select buffalo donors to enroll in embryo production programs, laying the basis for further investigations.
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
authors.forename: Aixin
authors.initials: A
authors.lastname: Liang
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Salzano
authors.affiliation: Institute of Genetics and Biophysics ABT, National Research Council, Naples, Italy; IRCCS Neuromed, Pozzilli, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: D'Esposito
authors.affiliation: Department of Food Science, University of Udine, Udine, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Comin
authors.affiliation: Department of Food Science, University of Udine, Udine, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Montillo
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
authors.forename: Liguo
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Campanile
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy. Electronic address: bgasparr@unina.it.
authors.forename: Bianca
authors.initials: B
authors.lastname: Gasparrini
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers; D002020:Buffaloes / Q000196:embryology / Q000502:physiology*; D004624:Embryo Transfer; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry*; D005786:Gene Expression Regulation / Q000502:physiology; D006107:Granulosa Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011974:Receptors, LH / Q000235:genetics / Q000378:metabolism*; D018000:Receptors, Peptide / Q000235:genetics / Q000378:metabolism*; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger; D011974:Receptors, LH; D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; C084621:anti-Mullerian hormone receptor; D054304:Anti-Mullerian Hormone
keywords: Anti-Mullerian hormone; Buffalo; Gene expression; Granulosa cell