Protocolo para la citología vaginal directa de ratas de laboratorio / Protocol for direct vaginal cytology of laboratory rats

Introducción: La citología vaginal directa es un método muy utilizado para la evaluación del ciclo estral de las ratas de laboratorio, pero la información acerca de los procedimientos e interpretación de los resultados aparece disgregada en la literatura, lo cual dificulta su empleo en los estudios de reproducción. Objetivo: Proponer un protocolo para la realización de la citología vaginal directa de ratas de laboratorio y la interpretación de los resultados. Material y métodos: Se combinó la información de la literatura y la experiencia de 10 años de estudios de reproducción, en los que se han empleado un total de 250 ratas Wistar hembras, siguiendo preceptos éticos establecidos. Se describen los procedimientos para la obtención de las muestras, mediante lavado vaginal, así como para su observación y análisis en estado húmedo, con microscopio óptico y cámara digital acoplada. Resultados: Se describen los tipos celulares principales presentes en el lavado vaginal y las características que permiten identificar cada fase o estado de transición del ciclo estral. Se discuten aspectos a considerar en la interpretación de los resultados, que incluye la relación con los cambios hormonales, los cuidados en la obtención de la muestra y la influencia de factores ambientales. Se muestran imágenes y figuras representativas para ilustrar el texto. Conclusiones: El trabajo constituye un protocolo para el estudio del ciclo estral de ratas de laboratorio, mediante la citología vaginal directa. Provee métodos no invasivos, sencillos y económicos, así como conocimientos esenciales para la interpretación de los resultados, que integran una guía de gran utilidad para los estudios experimentales de reproducción(AU)

Introduction: Direct vaginal cytology is a widely used method for the evaluation of the estrous cycle of laboratory rats, but the information about the procedures and interpretation of results is dispersed through literature, making its use difficult in investigations on reproduction. Objective: To propose a protocol for performing direct vaginal cytology of laboratory rats as well as for the interpretation of the results. Material and methods: The information obtained from literature and the experience of 10 years of investigation on reproduction in 250 female Wistar rats were combined following the established ethical principles. The procedures for obtaining samples by vaginal washing and by observation and analysis in humid state with light microscope equipped with digital camera were described. Results: The main types of cells present in the vaginal washing and the characteristics that allow us to identify each phase or transitional phases of the estrus cycle are described. Aspects to take into consideration in the interpretation of the results, which include the association between hormonal changes, the cares in the obtaining of the sample and the influence of environmental factors, are discussed. Representative images and figures are included to illustrate the text. Conclusions: The work consists of a study protocol of the estrus cycle of laboratory rats by direct vaginal cytology. It provides noninvasive, simple and cost-reducing procedures as well as essential knowledge for the interpretation of results which integrate a very useful guideline for experimental investigations on reproduction(AU)

Effects of ovarian varicose vein on mitochondrial structure, malondialdehyde and prooxidants: antioxidants balance in rat ovaries / Efectos de las venas ováricas varicosas sobre la estructura mitocondrial, niveles de malondialdehído y balance prooxidantes: antioxidantes en ovarios de ratas

Oxidative stress is increased in varicose veins. Many studies have implicated oxidative stress in the pathogenesis of infertility causing diseases of the female reproductive tract. The aim of this study was to determine whether varicocele can cause raised levels of reactive oxygen species and denaturation of mitochondrial structure in ovaries of female rats or not. In each experimental study, 15 weaning-age female rats were divided equally in 3 groups: Unilateral Varicose Vein (A), Sham (B) and Control (C) groups. Mitochondrial structure and malondialdehyde levels as a product of lipid peroxidation and Prooxidants-Antioxidants Balance were evaluated 60 days after intervention in proestrus stage. Comparisons between groups were made by the measured test. After 2 months, our results showed that mitochondrial structure ultra-structurally was denatured with histologic examination, malondialdehyde and prooxidants-antioxidants balance levels of left ovaries increased significantly in varicocele group compared to control and sham groups (P0.05). In the right side, malondialdehyde increased significantly, but in prooxidants-antioxidants balance levels, there is no significant differences between groups. The data of control and sham groups were the same. These findings may support the concept that increased levels of malondialdehyde and PAB in varicocele may cause negative effects on fertility, so using antioxidants maybe useful.

El estrés oxidativo aumenta en las venas varicosas. Diversas investigaciones han implicado al estrés oxidativo en la patogénesis de las enfermedades que causan la infertilidad del tracto reproductivo femenino. El objetivo de este estudio fue determinar si el varicocele puede provocar niveles de especies reactivas del oxígeno y la desnaturalización de la estructura mitocondriales en los ovarios de ratas. En cada estudio experimental, cinco ratas hembras en edad de destete se dividieron por igual en 3 grupos: Várices unilateral (A), simulado (B) y control (C). La estructura mitocondrial y los niveles de malondialdehído como un producto de la peroxidación lipídica y el balance pro-oxidantes-antioxidantes (BPA) se evaluaron 60 días después de la intervención en la etapa proestro. Las comparaciones entre grupos se realizaron mediante la prueba de medición. Después de 2 meses, observamos que la estructura mitocondrial se desnaturalizó ultraestructuralmente, los niveles malondialdehído y el balance prooxidantes-antioxidantes de los ovarios izquierdos aumentaron significativamente en el grupo A en comparación con los grupos B y C (P0.05). En el lado derecho, los niveles de malondialdehído aumentaron significativamente, pero el BPA, no mostró diferencias significativas entre los grupos. Los datos de los grupos B y C eran los mismos. Estos hallazgos pueden apoyar el concepto de que el aumento de niveles de malondialdehído y BPA en presencia de varicocele, puede causar efectos negativos sobre la fertilidad, por tanto el uso de antioxidantes puede resultar útil.

Efeitos da hiperprolactinemia sobre o útero de camundongos no proestro / Hyperprolactinemia effects on the female mice uterus during proestrous

OBJETIVO: avaliar o efeito da hiperprolactinemia induzida pela metoclopramida sobre o endométrio e miométrio de camundongos fêmeas na fase de proestro. MÉTODOS: 24 camundongos fêmeas foram divididas aleatoriamente em dois grupos: GCtr/controle e GExp/tratadas com metoclopramida (6,7 µg/g por dia). Após 50 dias, os animais foram sacrificados na fase de proestro, e o sangue foi coletado para determinação dos níveis de estradiol, progesterona e prolactina. Os cornos uterinos foram removidos e fixados em formol a 10 por cento; foram, então, processados para inclusão em parafina. Cortes de 4 µm foram corados pela hematoxilina-eosina (H/E). Na análise morfológica, foi utilizado microscópio de luz, da marca Carl Zeiss, com objetivas variando de 4 a 400 X, para caracterização de cada corte histológico. Na análise morfométrica, foi avaliada a espessura do epitélio superficial, da lâmina própria e do miométrio, com auxílio de um analisador de imagem (AxionVision, Carl Zeiss) acoplado ao microscópio de luz (Carl Zeiss). A análise estatística foi realizada pela ANOVA seguida pelo teste Wilcoxon. O valor de p foi considerado significante quando < 0,05. RESULTADOS: os achados mostraram aumento dos níveis séricos de prolactina (295,6±38,0 ng/mL) e redução significante dos de progesterona (11,3±0,9 ng/mL) no GExp em comparação ao GCtr (45,5±5,2 ng/mL e 18,2±1,6 ng/mL, respectivamente) (p<0,001). Com relação aos níveis séricos de estradiol, não foram obtidas diferenças significantes entre os grupos (GExp=119,1±12,3 pg/mL e o GCtr=122,7±8,4 pg/mL, p=0,418). O estudo morfológico mostrou que o útero do GExp apresentou endométrio com epitélio superficial e lâmina própria mais desenvolvida quando comparado ao GCtr, o mesmo ocorrendo no miométrio. Os valores morfométricos das espessuras do epitélio luminal (8,0±1,1 µm) e do endométrio (116,2±21,1x10² µm) do GCtr foram inferiores ao GExp (10,2±0,8 µm e 163,2±23,3x10² µm com p<0,05) respectivamente. Já os dados ...

PURPOSE: to evaluate the effect of hyperprolactinemia induced by metoclopramide on the endometrium and myometrium of female mice in the proestrus phase. METHODS: 24 female mice were randomly divided in two groups: CtrG/control and ExpG/treated with metoclopramide (6.7 mg/g daily). After 50 days, the animals were sacrificed in the proestrus phase, and the blood was collected to determine the levels of estradiol, progesterone and prolactin. The uterine horns were removed, fixed in 10 percent formaldehyde and processed before being included in paraffin. Slices of 4 µm were stained by hematoxylin and eosin (H/E). In the morphological analysis, a Carl Zeiss light microscope, with objectives varying from 4 to 400 X was used for each histological slice characterization. In the morphometrical analysis, the superficial epithelium, the lamina propria and the myometrium thickness were evaluated, with the help of an image analyzer (AxionVision - Carl Zeiss) attached to the light microscope (Carl Zeiss). The statistical analysis was done by ANOVA, followed by the Wilcoxon test. P-value was considered as significant, when <0.05. RESULTS: our findings have shown an increase in the seric levels of prolactin (295.6±38.0 ng/mL) and significant decrease in the progesterone levels (11.3±0.9 ng/mL) in the ExpG, as compared to the CtrG (45.5±5.2 ng/mL and 18.2±1.6 ng/mL, respectively; p<0.001). Concerning the seric level of estradiol, significant differences between the groups were not obtained (ExpG=119.1±12.3 pg/mL and CtrG=122.7±8.4 pg/mL; p=0.418). The morphological study has shown that the uterus from the ExpG presented the endometrium with more developed superficial epithelium and lamina propria, as compared to the CtrG, the same happening with the myometrium. The thickness morphometrical values of the luminal epithelium (8.0±1.1 µm) and endometrium (116.2±21.1x10² µm) from the CtrG were lower than the ones from the ExpG (10.2±0.8 µm and 163.2±23.3x10² ...