Gene expression of small leucine-rich proteoglycans (SLRPS), decidual prolactin and growth factors involved in decidualization of the endometrium of female mice with pseudoprenhez with hyperprolactinemia induced by metoclopramide
100 employees are female mice, adults, 90 days of age, weighing 30g, virgins and 34 mice vasectomized males, adults, 180 days of age, weighing 35g with approval by the Ethics Committee (Project No. 0809/04) Annex 1. In order to analyze the decidualised endometrium will be conducted two experiments: the induction of hyperprolactinemia and the state then the induction pseudoprenhez. Experimental I, females will be divided into two groups with 50 animals each, namely: control group (Ctr): subjected to daily injections subcutaneously 0.2 ml of saline for 50 days the experimental group (HPrl): subjected to daily injections subcutaneously of 6.7 mg/g of metoclopramide, dissolved in 0.2 ml of saline for 50 days. The induction of hyperprolactinemia will be held according to the protocol recommended by SINGTRIPOP et al., 1991. After 50 days shall be made vaginal smear test to subdivide the animals according to the estrous cycle, namely: group (Ctr: 10 animals/stage): proestrus, estrus, metaestrus and diestrus and group (HPrl: 10 animals/stage): proestrus, estrus, estrus, metaestrus and diestrus. Then the animals, after euthanasia by anesthetic depth associated with the muscle relaxant and the uterine horns are removed, processing described below. Experiment II, the females are in estrus phase will be placed for mating with males vasectomies, forming two groups (Ctr and HPrl) with 10 female mice each with pseudoprenhez in this period, the animals continued to receive treatments. On the 5th day of pseudoprenhez after proof will be done by deepening the anesthetic euthanasia associated with muscle relaxant and the uterine horns are removed. The uterine horns for the analysis of gene expression will be put into tubes with 2 ml of RNAlater for subsequent RNA extraction and quantification by the technique of Polymerase Chain Reaction Real-Time (PCR - Real Time) and the uterine horns for immunohistochemistry to be fixed for 24 hours in 10% formaldehyde diluted in PBS pH 7.4 phosphate buffer.
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