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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2021.tde-21072021-074922
Document
Author
Full name
Loren D'Aprile
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2021
Supervisor
Committee
Keohane, Paula de Carvalho Papa (President)
Campos, Danila Barreiro
Garcia, Rita de Cássia Maria
Title in Portuguese
A importância da família dos fatores de crescimento endotelial vascular na reprodução
Keywords in Portuguese
Cadelas
Corpo lúteo
Fatores angiogênicos
Isoformas
VEGFA
Abstract in Portuguese
O ciclo estral da cadela difere de fêmeas de outras espécies e tem como protagonista principal o corpo lúteo, glândula endócrina transitória que secreta diversos fatores relacionados à sua manutenção e regressão. Um dos fatores chaves que regula a angiogênese é o VEGFA, fator de crescimento endotelial pertencente a uma ampla família. O objetivo do trabalho foi revisar a importância do VEGFA, de suas isoformas, seus receptores e dos fatores de crescimento na angiogênese, na reprodução e no corpo lúteo canino. Sobre as isoformas angiogênicas do VEGFA e sua estruturação em exons e introns, nada se achou na espécie canina. Sugerimos a estrutura das isoformas caninas homologas às do ser humano para posterior validação. Adicionalmente, identificamos a expressão de fatores angiogênicos durante o diestro não gestacional na espécie canina, através da técnica de RNA-seq. Para tal, foram utilizados corpos lúteos provenientes de 18 cadelas que passaram por ovariosalpingohisterectomia (OSH) nos dias 10, 20, 30, 40, 50 e 60 após a ovulação (p.o). Os corpos lúteos coletados foram utilizados para extração de RNA total a partir do protocolo de TRIZOL e posterior análise por RNA- seq, seguindo o protocolo TruSeq RNA Sample Preparation Guide, descrito por Illumina. Identificamos a expressão diferencial do VEGFA, VEGFC, VEGFD, VEGFR1, VEGFR2 e VEGFR3 nas comparações entre o período luteotrófico e regressão luteínica. Os dados demonstram maior expressão de fatores angiogênicos no dia 20 p.o, período relacionados à proliferação celular e manutenção da função do corpo lúteo canino.
Title in English
The importance of vascular endothelial growth factor family in reproduction
Keywords in English
Angiogenic factors
Bitches
Corpus luteum
Isoforms
VEGFA
Abstract in English
The estrous cycle of bitches differs from that of other species and its main protagonist is the corpus luteum, a transient endocrine gland that secretes several factors related to its maintenance and regression. One of the main factors that regulate angiogenesis is VEGFA, an endothelial growth factor belonging to a large family. The aim of the study was to shed light on the importance of VEGFA, its isoforms and receptors in angiogenesis, reproduction and canine corpus luteum. The angiogenic isoforms of this growth factor have been investigated for its structure in exons and introns, and nothing has been found about this conformation in dogs. Here we suggest the structure of canine isoforms homologous to that of humans for further validation and we seek to identify the expression of angiogenic factors during non- gestational diestrus in the canine species, using the RNA-seq technique. For this purpose, corpus luteum of 18 bitches submitted to ovariosalpingohisterectomy (OSH) were used on days 10, 20, 30, 40, 50 and 60 after ovulation (p.o). The collected corpus luteum were used for extraction of total RNA using the TRIZOL protocol and subsequent analysis by RNA-seq, following the protocol TruSeq RNA Sample Preparation Guide, described by Illumina. We identified the differential expression of VEGFA, VEGFC, VEGFD, VEGFR1, VEGFR2 and VEGFR3 when comparing the luteotrophic and regression periods. The data demonstrate the active participation of angiogenic factors on day 20 post ovulation, a period related to cell proliferation and maintenance of canine corpus luteum function.
 
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Publishing Date
2021-08-16
 
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