17b-estradiol role on the luteolytic process in non-pregnant dogs

• Grant number: 11/17768-3
• Support Opportunities: Regular Research Grants
• Duration: February 01, 2012 - July 31, 2014
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Paula de Carvalho Papa Keohane
• Grantee: Paula de Carvalho Papa Keohane
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The corpus luteum (CL) is a temporary endocrine gland, which goes trought different developmental phases: formation, maintenance and regression and reaches full secretory capacity when formation is complete. Steroidogenic profile differs over diestrus : progesterone (P4) is markedly elevated in the formation period; decreasing P4 and increasing 17b-estradiol (E2) are observed during maintenance and E2 highest diestrous levels on day 40 post ovulation (p.o.). Nevertheless, mechanisms regulating CL function and life span are not compeltely elucidated and show species specific differences. Special interest relies on the luteolytic phase, or regression phase, as some authors prefer when talking about the non-pregnant bitch, a moment that, differently from other species, PGF2a plays no important role. We hypothesize that E2 is one of the triggers for the non-pregnant dog luteolysis and that its variations over diestrus contribute to the establishement of metabolic alterations, specially insulin resistance. To test this hypothesis 24 non-pregnant bitches will be submitted to ovariosalpingohysterectomy on days 10, 20, 40, 60, 80 and 100 p.o. (n = 4 per group). Blood samples will be collected prior to anesthesia for hormonal measurements of P4, E2, insulin, prolactin, GH and LH and glycaemia will assessed as a metabolic parameter. Some CL will be frozen in liquid nitrogen for posterior gene and protein expression evaluation as microarray, real time PCR and western blotting. For immunhistochemistry CL samples will be fixed in 4% paraformaldehyde. Remaining CL will be transported to the laboratory in phospahte buffer for cell culture and E2 and/or E2 receptor blocker experiments. Genes involved in apoptosis, prolieferation and steroidogenesis, specially E2 and P4 receptors will be analyzed after cell treatments. Samples undergoing microarray analysis will be used to define other candidate genes related to luteal regression and metabolic alterations. At least 4 genes related to CL regression and 4 to metabolic alterations which have been differentially expressed will be validated through real time PCR, western blotting and immunhistochemistry. Data will be analyzed by MinitabÒ statistical program and differences will be considered when p < 0.05. (AU)

Scientific publications

(4)

(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)

PAPA, PAULA DE CARVALHO; MEDEIROS DE CARVALHO SOUSA, LIZA MARGARETH; SILVA, RENATA DOS SANTOS; DE FATIMA, LUCIANA ALVES; DA FONSECA, VANESSA UEMURA; DO AMARAL, VANESSA COUTINHO; HOFFMANN, BERND; ALVES-WAGNER, ANA BARBARA; MACHADO, UBIRATAN FABRES; KOWALEWSKI, MARIUSZ PAWEL. Glucose transporter 1 expression accompanies hypoxia sensing in the cyclic canine corpus luteum. Reproduction, v. 147, n. 1, p. 81-89, JAN 2014. (10/07373-9, 11/17768-3)

FATIMA, L. A.; EVANGELISTA, M. C.; SILVA, R. S.; CARDOSO, A. P. M.; BARUSELLI, P. S.; PAPA, P. C.. FSH up-regulates angiogenic factors in luteal cells of buffaloes. DOMESTIC ANIMAL ENDOCRINOLOGY, v. 45, n. 4, p. 224-237, NOV 2013. (11/17768-3)

MONA E PINTO, J.; PAVANELO, JR., V.; ALVES DE FATIMA, L.; MEDEIROS DE CARVALHO SOUSA, L. M.; PACHECO MENDES, G.; MACHADO FERREIRA, R.; AYRES, H.; SAMPAIO BARUSELLI, P.; PALMA RENNO, F.; DE CARVALLO PAPA, P.. Treatment with eCG Decreases the Vascular Density and Increases the Glandular Density of the Bovine Uterus. REPRODUCTION IN DOMESTIC ANIMALS, v. 49, n. 3, p. 453-462, JUN 2014. (11/17768-3)

MEDEIROS DE CARVALHO SOUSA, LIZA MARGARETH; SILVA, RENATA DOS SANTOS; DA FONSECA, VANESSA UEMURA; LEANDRO, RAFAEL MAGDANELO; DI VINCENZO, THIAGO SENNA; ALVES-WAGNER, ANA BARBARA; MACHADO, UBIRATAN FABRES; PAPA, PAULA DE CARVALHO. Is the canine corpus luteum an insulin-sensitive tissue?. Journal of Endocrinology, v. 231, n. 3, p. 223-233, DEC 2016. (11/17768-3, 10/07373-9)