2011 (SBTE) 25th Annual Meeting Proceedings - International ...

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011.
A187 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION
ESTROGEN SOURCES IN FEMALE SIRIO HAMSTERS (
(MESOCRICETUS AUR
URATUS
TUS) ) AFTER OVARIET
ARIETOMY
Kelly Annes 1 , Marie Odile M. Chelini 2 , Nivea Lopes Souza 3 , Silvia Renata Gaido Cortopassi 3 & Marcella Pecora Milazzotto 1
1
CENTRO DE CIÊNCIAS NATURAIS E HUMANAS - UFABC, SANTO ANDRÉ, SP, BRAZIL. 2 INSTITUTO DE PSICOLOGIA - USP, SÃO PAULO, SP, BRAZIL. 3 FACULDADE DE MEDICINA
VETERINÁRIA E ZOOTECNIA - USP, SÃO PAULO, SP, BRAZIL.
The technique of doses of steroid hormones in feces is increasingly used in animals of small size, a characteristic that makes it
difficult to obtain blood samples. In a previous work estrogen and progesterone were measured in feces and blood of non-treated females (IN)
and ovariectomized (OV). Although quantitative differences between the methods (there was no difference between the levels of fecal estrogen
groups, since the serum concentrations of estradiol were different), the possibility of detection of these metabolites in the feces is shown in the
study of relevant reproductive physiology these animals. Based on this data, the hypothesis of this work is that although female Sirio hamsters
(Mesocricetus auratus) OV produce estrogen in non-ovarian tissues which cannot be detected in sistemic circulation, these metabolites may be
quantified in their feces. Therefore, our objective was to verify the presence and quantify estrogen metabolites in feces of non-treated females
(IN) or OV and quantify the gene expression of the responsible for the synthesis of aromatase (ARO) in non-ovarian tissues (brain, liver, adrenal
glands and gastric mucosa). For such, 11 adult females were randomly divided in two groups: OV (n = 6) and IN (n = 5). Estrogen
concentrations from the OV and IN animals was followed for 7 months after the ovariectomia. After this period the females were euthanized for
the target tissue gathering for the analysis of the ARO expression by RT-PCR. The collect of feces were made before the OV of 4 on 4 h for 4
days, the day of OV and 8 days later (once a day), from 30 days and every 30 days (produced by 24 for 6 months ). Estrone, estriol, estradiol
was extracted by suspension in methanol 80% and quantified by enzyme immunoassay. The analysis of the ARO expression showed that there
was no difference on this enzyme expression in the brain in OV or in IN, there was expression in the liver and gastric mucosa, nevertheless, the
expression in adrenals was greater in OV. Before surgery, the 11 females showed fecal metabolites curve exactly parallel the dosage blood, with
6 h delay. The short and long term after surgery, OV females showed marked and steady decline of fecal estrogens. After challenge with ACTH
dosage of the curve was similar of non-treated females obtained before surgery. We conclude that the adrenal has become a major source of
estrogen in females OV, and these metabolites in the feces of these animals. However, further studies would be required for the determination of
the majority of estrogen metabolite detected in feces. [Acknowledgments: FAPESP process 2009/ 52750-8].
Keywords: hamster, oestrogen, ovariectomy.
A188 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION
IMMUNOLOC
OCALIZA
ALIZATION OF THE PIDERMAL GROW TH FACT
CTOR (EGF), TRANSFORMING GROW TH FACT
CTOR-
ALPHA (TGF
GFA) AND EGF RECEPTOR (EGFR) IN THE OVAR
ARY OF THE BITCH
Diogo José Cardilli 1 , José Félix Pérez-Gutiérrez 2 , Kellen De Sousa Oliveira 1 , Carolina Franchi João 3 , Fabiana Azevedo Voorwald 1 ,
J oão Ademir Oliv
liveir
eira 1 & Gilson Hélio Toniollo
1
1
FCAV/UNESP, JABOTICABAL, SP, BRAZIL. 2 UCM-MADRID, MADRID, ESPANHA. 3 UFP-CASTANHAL, CASTANHAL, PA, BRAZIL.
The EGF and the TGFa, belong to the same family. They exhibit 40-50% structural homology; both bind to the EGFr and induce
similar biological effects: cellular division, differentiation and survival. Previous studies had shown their role in a wide variety of physiological
functions including oocyte maturation in different species such as: porcine, rodents, primates, bovine and canine; however their localization in
the ovary of the bitch has not been examined. The aim of this study was to determine the distribution of EGF, TGFa and EGFr in the ovary of
the bitch. Reproductive tracts from 34 bitches at the different phases of the estrous cycle were collected, fixed and processed with paraffin
embedding. Sections were cut, mounted, dried, deparaffinized and rehydrated. Immunocytochemistry was performed using a 1:20 dilution of the
following antisera: anti-EGF-pc (Ab-3, Calbiochem), Anti-TGFá-mc (Ab-2) and anti-EGFr receptor-mc (Ab-5). Samples were incubated
overnight. Primary antibody binding was detected using a biotynilated secondary goat-anti-rabbit or rabbit anti-mouse IgG with avidin-biotinperoxidase
complexing. The detection reagent was AEC (Vector). Immunohistochemistry revealed the presence of EGF, TGFα and EGFr in the
ovary of the bitch. Within the cell, both ligands, EGF and TGFá were localized in the cytoplasm, while EGFr staining was nuclear. EGF
immunolocalized in the follicular cells (FC) and in the oocytes of secondary and tertiary follicles, as well as, in the vascular endothelium,
granulosa cells strings and in the luteal cells (LC). Positive reaction to TGF-α was found in the superficial epithelium, stroma, cortical tubules,
vascular endothelium, LC, as well as, in the FC and oocytes of primary, secondary and tertiary follicles. EGFr was immunolocalized in the
superficial epithelium, stroma, cortical tubules, vascular endothelium, LC and FC of primary, secondary and tertiary follicles. This study
describes the localization of EGF system in the ovary of the bitch and suggest its participation in the regulation of ovarian functions such as
follicular growth and luteinization.
Keywords: stroma, immunocytochemistry, bitches.
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