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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2007.tde-13122007-150929
Document
Author
Full name
José Sergio de Arruda Gonçalves
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Visintin, José Antonio (President)
Crusco, Silvia Edelweiss
Vannucchi, Camila Infantosi
Title in Portuguese
Análise da expressão gênica diferencial dos receptores de estrógeno (erα e erβ) e progesterona (pr) em oócitos e células do cumulus nas diferentes fases do ciclo estral em cães
Keywords in Portuguese
Cães
Cumulus
Estrógeno
Oócito
Progesterona
Receptores
Abstract in Portuguese
O objetivo do presente estudo foi analisar a expressão gênica dos receptores de estrógeno (ERα e ERβ) e progesterona (PR) em oócitos e células do cumulus de cadelas nas diferentes fases do ciclo estral. Ovários de cadelas previamente avaliadas e submetidas à ovário-histerectomia foram fatiados, sob refrigeração, em PBS com 10% de SFB. No momento imediatamente pré-cirúrgico foi colhido sangue da cadela, que foi submetido posteriormente à dosagem de estradiol e progesterona. As cadelas foram agrupadas nas diferentes fases do ciclo estral de acordo com a avaliação prévia por colpocitologia, colposcopia e dosagem sérica de progesterona. Os oócitos recuperados após o fatiamento foram denudados mecanicamente. Oócitos e células do cumulus foram criopreservados separadamente e posteriormente foram submetidas a PCR em tempo real para quantificação relativa dos genes de interesse. A expressão gênica de ERα, ERβ e PR em oócitos e células do cumulus não foi diferente nas fases do ciclo estral. Foi possível, pela primeira vez, relatar a presença de ERα e ERβ em oócitos de cadelas.
Title in English
Analysis of differential gene expression of estrogen (erα and erβ) and progesterone (pr) receptors in oocytes and cumulus cells on different phases of the estrous cycle in dogs
Keywords in English
Cumulus
Dogs
Estrogen
Oocyte
Progesterone
Receptors
Abstract in English
The objective of the present study was to analyze the gene expression of estrogen (ERα and ERβ) and progesterone (PR) receptors in oocytes and cumulus cells on different phases of the estrous cycle in dogs. Ovaries from selected bitches were recovered after ovary-hysterectomy and sliced at low temperatures in PBS supplemented with 10% FCS. Immediately after surgery, blood samples were harvested for measurement of estrogen and progesterone serum concentration. Bitches were grouped within different phases of the estrous cycles, according to previous evaluation of colpocytology, colposcopy and serum progesterone levels. Oocytes recovered after slicing were mechanically denuded. Isolated oocytes and cumulus cells were cryopreserved and submitted to RT-PCR followed by a real time PCR for relative quantification of gene expression. Gene expression of ERα, ERβ and PR in oocytes and cumulus cells was not different among phases of the estrous cycles. For the first time it was possible to demonstrate ERα and ERβ in dog oocytes.
 
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Publishing Date
2007-12-19
 
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