Characterization of anti-Mullerian hormone concentrations - AMH in vivo and in vitro in bitches and gray wolves (Canis lupus baileyi)

• Grant number: 15/09246-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: October 01, 2015
• End date: July 15, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Maria Denise Lopes
• Grantee: Camila Louise Ackermann
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated scholarship(s): 15/24963-8 - Characterization of anti-Mullerian hormone concentrations - AMH in vivo and in vitro in bitches and gray wolves (Canis lupus baileyi), BE.EP.PD

Abstract

We aim to characterize anti-Mullerian hormone concentrations - AMH in vivo and in vitro in bitches and gray wolves (Canis lupus baileyi). Two experiments will be perform. Experiment 1: Determination of serum hormone levels of anti-muleriano hormone in female dogs and gray wolves: Ten females of grey wolf, from International Wolf Center (Minessota, EUA) will be used as an experimental model for Mexican Grey Wolves. Two blood samples will be collected from each wolf twice, one during their reproductive season (winter) and out off it (summer). The number of animals to be collected may be increase. For domestic dogs, 45 bitches (placed in 3 different groups: pre-pubertal, pubertal and old) will have a single blood sample collected. All samples will be centrifuged and serum stored at -20ºC until assays be performed. AMH and estrogen will be mensured. Data will be submitted to statistical tests. Experiment 2: AMH production in fresh and thawed dog ovaries during in vitro culture. At least 20 bitches will be spayed and will have a blood sample collected for future hormonal assay. Ovaries will be isolated, and stored in containers at 37 ºC. Ovaries without CL will have their cortex sliced in 8 pieces. Three fresh pieces will be used for in vitro culture and one will be use to histological evaluation. The other 4 pieces will be submitted to cryopreservation, using the same method used for Mexican Grey Wolves in Saint Louis Zoo. The slices will be thawed, for that the pieces will be immersed in a Petri dish containing thawing solution thereafter it will be transferred to a petri dish containing Sucrose solution. Both fresh and thawed samples will be cut into smaller pieces and placed in 96 well plate, witch well containing 400 µL of growth media. Plates will be maintained at 5% CO2 and 38.5ºC. Every other day 200 µL of media will be removed and stored at -20oC for AMH, P4 and estrogen assay, the same volume will be replaced with fresh media. After 7 days of culture, sample will be removed and processed to histological analysis. Hormonal assay will be performed to determine estrogen, AMH and P4 from in vitro culture media and serum. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ACKERMANN, C. L.; ASA, C. S.; KRISHER, R.; BAUMAN, K.; CASEY, S.; LOPES, M. D.. Evaluation of follicular growth and tissue viability in vitrified/warmed domestic dog ovaries after in vitro culture. REPRODUCTION IN DOMESTIC ANIMALS, v. 52, n. 2, SI, p. 77-81, APR 2017. (13/18337-1, 15/09246-8, 15/24963-8)

ACKERMANN, C. L.; ASA, C. S.; KRISHER, R.; BAUMAN, K.; CASEY, S.; LOPES, M. D.. Evaluation of follicular growth and tissue viability in vitrified/warmed domestic dog ovaries after in vitro culture. REPRODUCTION IN DOMESTIC ANIMALS, v. 52, p. 5-pg., 2017-04-01. (15/24963-8, 15/09246-8, 13/18337-1)