Carina Verna

Concentration and distribution of hyaluronic acid in mouse uterus throughout the estrous cycle

OBJECTIVE To quantify and study the immunoexpression of hyaluronic acid (HA) in the uterine horns of the mouse throughout the estrous cycle phases. DESIGN Experimental study using an ELISA-like fluorometric assay to quantify HA and an avidin-biotin immunoperoxidase method using biotinylated hyaluronan-binding protein for histochemical studies. SETTING University-based laboratory. ANIMAL(S) Forty regularly cycling adult female mice were divided into four groups according to the diagnosed phase of the cycle: proestrus, estrus, metaestrus, and diestrus. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Histologic samples of the uterine horns. Immunohistochemical reaction was evaluated by detection of HA and CD44 distribution within the uterine horn. Tissue HA content was determined through an ELISA-like fluorometric assay with the same hyaluronan-binding protein and with europium-labeled streptavidin. RESULT(S) The immunohistochemical HA and CD44 reactions were most intense during diestrus, mainly below the luminal epithelium. HA was strongly stained in the connective tissue near the myometrium layer during metaestrus. The biochemical data showed that the highest concentration of HA in uterine horns occurred during diestrus (4053.0 +/- 651.4 ng/g dry tissue) compared with other phases. CONCLUSION(S) Our data show that the expression of HA in mouse uterine horns is highest during the diestrous phase. The fluctuations of HA in the mouse uterus during the estrous phase may be related to the varying estrogen and P levels during the cycle and may be important as far as embryo implantation is concerned.

Effects of metoclopramide on the mouse anterior pituitary during the estrous cycle

The symptoms of hyperprolactinemia in women mainly result from changes in the release of gonadotropins and the consequent repercussions on ovarian function.1-3 Metoclopramide is a hyperprolactinemic drug that is used as an antiemetic agent.4,5 In female mice, metoclopramide-induced hyperprolactinemia causes changes in the reproductive system, mainly in ovarian function and the endometrium.6-10 While there is extensive data regarding prolactin regulation and hyperprolactinemic states, the direct effects of metoclopramide on the morphological and functional aspects of pituitary cells remain unknown. Therefore we have evaluated the histomorphological and immunohistochemical changes in lactotrophs of female mice upon treatment with metoclopramide.

Efeito da hiperprolactinemia induzida pela metoclopramida na glândula lacrimal: estudo experimental

PURPOSE: to evaluate the morphological changes in murine lacrimal glands by metoclopramide-induced hyperprolactinemia during the proestrus phase or pregnancy. METHODS: forty adult mice were divided into two groups: CTR1 (control) and MET1 (treated with metoclopramide). After fifty days, half of the mice were sacrificed. The remaining animals were mated, and then labeled as pregnant controls (CTR2). Part of these animals were treated with metoclopramide and constituted the metoclopramide-treated pregnant (MET2) group. The CTR2 and MET2 groups were sacrificed on the 6th day of pregnancy. The blood was collected for determination of the hormonal levels of estradiol and progesterone by a chemoluminescent method. The lacrimal glands were then removed, fixed in 10% formaldehyde and stained with HE. The morphometric analysis was performed using the Axion Vision program (Carl Zeiss) to measure acinar nuclear and cellular volumes. RESULTS: the nuclear and cellular volumes of the lacrimal glands in the MET1-(152.2±8.7; 6.3±1.6 µm3) and MET2-(278.3±7.9; 27.5±0.9 µm3) treated groups were lower than those in CTR1 (204.2±7.4; 21.9±1.3 µm3) and CTR2 (329.4±2.2; 35.5±2.0 µm3), respectively. There was a significant hormonal level reduction in the animals that received metoclopramide compared to controls (CTR1: estradiol = 156.6±42.2 pg/ml; progesterone = 39.4±5.1 ng/ml; MET1: estradiol = 108.0±33.1 pg/ml; progesterone = 28.0±6.4 ng/ml; CTR2: estradiol = 354.0±56.0 pg/ml; progesterone = 251.0±56.0 ng/ml; MET2: estradiol = 293.0±43.0 pg/ml, progesterone = 184.0±33.0 ng/ml). CONCLUSION: metoclopramide-induced hyperprolactinemia produced morphological signs of reduction of cellular activity in lacrimal glands during the proestrus phase and pregnancy. It is hypothesized that this effect might be related to the hyperprolactinemia-induced decrease in the hormonal production of estrogen and progesterone.

Concentration of glycosaminoglycan in ovariectomized mice uterus after treatment with ovarian steroids.

Abstract The aim of this study was to evaluate the amount of non- and sulfated glycosaminoglycans in the ovariectomized mice uterus, after treatment with ovarian steroids. For this purpose, 50 adult female mice were divided into five groups with 10 animals/each: control group: CG (ovary intact), and ovariectomized groups: OG (vehicle), EG (estradiol), PG (progesterone) and EPG (estradiol combined to progesterone). The treatments started 30 days after ovariectomy. All the animals were treated for 50 consecutive days. These hormones were administered in a sterile oily solution via gavage. Twenty-four hours after the last treatment, all animals were euthanized, removing the uterine horn for biochemical analyses. To quantify, the hyaluronic acid (HA) used ELISA-like fluorometric assay, and the sulfated glycosaminoglycans (GAGs) used agarose gel electrophoresis. The amount of HA was significantly higher in the group treated with progesterone (PG) compared to the others groups (p < 0.05), and in the group treated with estradiol (EG), the amount of chondroitin/dermatan sulfate was significantly higher compared to the others groups (p < 0.05), and in the group treated with progesterone (PG), the amount of heparan sulfate was significantly lower compared to the others groups, except to control group (p < 0.05). Our results showed that the estroprogestative therapy after long time (50 days) profoundly affected the amount of glycosaminoglycans in uterine. These changes may be indicative of uterine pathology such as the development of tumor.

Influence of hyperprolactinemia on collagen fibers in the lacrimal gland of female mice

OBJECTIVE: To quantify the collagen fibers in the lacrimal gland of female mice with hyperprolactinemia. METHODS: Forty adult female mice were randomly divided into two groups with 20 animals each: nonpregnant control (CTR1, control group, 0.2 mL of saline solution) and nonpregnant experimental (HPRL1, experimental group, 200 µg/day metoclopramide). Treatments lasted for 50 consecutive days. On day 50, 10 females from each group (control and experimental) were euthanized in the proestrus phase; then, the blood was collected and the lacrimal glands were removed. Thereafter, the remaining females were placed with the mates and continued to receive treatment with saline solution or metoclopramide. On the 6th post-coital day, 10 pregnant females from the control group (CTR2) and 10 pregnant females from the experimental group (HPRL2) were euthanized, after which blood was collected and the lacrimal glands removed. The lacrimal glands were processed for morphological analyses and collagen quantification, and prolactin and sex steroid levels were measured in the blood samples. Data were statistically analyzed using an unpaired Student t test (p<0.05). RESULTS: Morphological analysis revealed greater structural tissue disorganization of the lacrimal glands in the metoclopramide-treated groups. The total collagen content was significantly higher in the HPRL1 group than in the CTR1 group (p<0.05), whereas the difference between the CTR2 and HPRL2 groups was not significant. CONCLUSION: Our data suggest an impairment in the functioning of the lacrimal gland as a consequence of increased prolactin levels and decreased serum levels of estrogen and progesterone.

Molecular features of sexual steroids on cartilage and bone

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.

Aspectos moleculares dos esteroides sexuais sobre a cartilagem e os ossos

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.

Efeito da hiperprolactinemia induzida pela metoclopramida na córnea de camundongas

PURPOSE: To evaluate the morphological changes in murine cornea upon metoclopramide-induced hyperprolactinemia during the proestrous phase or pregnancy. METHODS: Forty adult mice were divided into two groups: (control) CTR1 and (treated with metoclopramide (MET1). After fifty days, half of the mice were sacrificed. The remaining animals were mated, and then labeled as pregnant controls (CTR2). Part of these animals were treated with metoclopramide and constituted the metoclopramide-treated pregnant (MET2) group. The groups CTR2 and MET2 were sacrificed on the 6th day of pregnancy. The hormonal levels were assessed by chemioluminescence and radioimmunoassay methods and the cornea was removed for the histomorphometric study. RESULTS: The epithelial, stromal, endothelial and total thickness in the experimental group was: MET1 and MET2 were higher than one in the control group: CTR1 and CTR2. There was a significant reduction of the hormonal level in the animals that received metoclopramide as compared to controls (CTR1: estradiol = 156.6±42.2 pg/ml; progesterone = 39.4±5.1 ng/ml; prolactin = 130.4±26.2 ng/ml; MET1: estradiol = 108.0±33.1 pg/ml; progesterone = 28.0±6.4 ng/ml; prolactin = 551.5±23.3 ng/ml; CTR2: estradiol = 354.0±56.0 pg/ml; progesterone = 251.0±56.0 ng/ml; prolactin = 423.2±28.1 ng/ml; MET2: estradiol = 293.0±43.0 pg/ml; progesterone = 184.0±33.0 ng/ml; prolactin = 823.1±51.1 ng/ml). CONCLUSION: The metoclopramide-induced hyperprolactinemia may increase corneal layers, mainly in pregnant mice. Possibly, this effect is related to reduction in estrogen and progesterone production.

Microscopic alterations in silicone tubes surface after application of ophthalmological lubricants

Objective: To identify microscopic morphological alterations in the surface of silicone tubes used for intubation of the lachrymal system after exposure to ophthalmological lubricants. Methods: Experimental, descriptive and longitudinal study consisted of the application of ophthalmological lubricants in silicone tubes. The tubes were divided in: Group 1 (Cylocort®), 2 (Epitezan®), 3 (Labcaina®), 4 (Liposic®), 5 (Maxinom®) and 6 (Vista Gel®). One tube was not exposed to any lubricant, used as control. The tubes were observed and photographed after 2 hours, 30 days, 45 days before and after cleaning the surface and lumen. The following aspects were observed: surface (regularity, transparency, quantity, size and shape of the substances) and lumen (obstruction). Results: Control: irregular surface with pores after 2 hours: Group 1 – irregular surface with presence of film; Groups 2, 3 and 5 – abundant and irregular quantity of ointment at the surface; Group 4 – discrete modification at the surface; Group 6 – growth of pigmented (brownish) structures with filaments in the lumen, with discrete film in the surface. 30 Days: Groups 1, 4 and 5 – increase of the irregular superficial film; Group 2 – crust with notorious horizontal lines; Group 3 – diminution of the superficial film; Group 4 – crust less evident. Group 6 – increase of the structure seen with 2 hours of exposition, arboriform aspect. Forty-five days pre cleaning: Group 4 – diminution of the surface crust; Group 6 – expansion of the arboriform structure; unaltered findings in other groups. 45 days after cleaning: Groups 1 and 5 – light diminution of the surface crust; Groups 2, 3 and 4 – kept the modifications; Group 6 – the structure inside the lumen was not identified, clear surface, without evidence of film. Conclusions: Microscopic morphological alterations in the surface and lumen of silicone tubes can occur when those remain in contact with determined ophthalmological lubricants.