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      Determination of the estrous cycle phases of rats: some helpful considerations Translated title: Determinação das fases do ciclo estral em ratas: considerações úteis

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          Abstract

          The short length of the estrous cycle of rats makes them ideal for investigation of changes occurring during the reproductive cycle. The estrous cycle lasts four days and is characterized as: proestrus, estrus, metestrus and diestrus, which may be determined according to the cell types observed in the vaginal smear. Since the collection of vaginal secretion and the use of stained material generally takes some time, the aim of the present work was to provide researchers with some helpful considerations about the determination of the rat estrous cycle phases in a fast and practical way. Vaginal secretion of thirty female rats was collected every morning during a month and unstained native material was observed using the microscope without the aid of the condenser lens. Using the 10 x objective lens, it was easier to analyze the proportion among the three cellular types, which are present in the vaginal smear. Using the 40 x objective lens, it is easier to recognize each one of these cellular types. The collection of vaginal lavage from the animals, the observation of the material, in the microscope, and the determination of the estrous cycle phase of all the thirty female rats took 15-20 minutes.

          Translated abstract

          Em razão da curta duração de seu ciclo estral, ratas constituem bom modelo para o estudo das alterações que ocorrem durante o ciclo reprodutivo. O ciclo estral de ratas tem duração média de quatro ou cinco dias e é caracterizado por quatro fases, proestro, estro, metaestro e diestro, as quais podem ser determinadas pelos tipos celulares observados no esfregaço vaginal. Como a coleta da secreção vaginal e o uso de material corado geralmente são demorados, o objetivo deste trabalho foi fornecer aos pesquisadores algumas considerações úteis para a determinação das fases do ciclo estral de maneira rápida e prática. A secreção vaginal de trinta ratas foi coletada diariamente pela manhã, durante um mês, e o lavado vaginal não corado foi observado a fresco, em microscópio ótico, sem a utilização das lentes do condensador. O uso das objetivas de aumento de 10 vezes facilita a análise da proporção entre os três tipos celulares presentes no material. O uso das objetivas de aumento de 40 vezes permite o reconhecimento de cada um dos tipos celulares. Descrevemos o método utilizado, no qual a coleta do lavado vaginal, a observação do material e a identificação das fases do ciclo estral de 30 ratas podem ser realizados em 15-20 minutos.

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          The control of progesterone secretion during the estrous cycle and early pseudopregnancy in the rat: prolactin, gonadotropin and steroid levels associated with rescue of the corpus luteum of pseudopregnancy.

          The hormonal factors associated with converting a corpus luteum of estrous cycle into a corpus luteum of pseudopregnancy were studied by measuring LH and FSH prolactin, estradiol and progesterone levels in decapitated rats during the 4-day estrous cycle and a comparable time of pseudopregnancy (lights on 0600-0800 hr.). During the estrous cycle, prolactin, LH and FSH remained low and unchanging except on the afternoon of proestrus, when typical proestrous surges were observed. In contrast, estradiol levels began to increase on D-1, from baseline values of 7 pg/ml to approximately 15-20 pg/ml. These levels were maintained until the afternoon of D-2 when estradiol further increased to reach peak levels of 40-50 pg/ml by 0900 hr on proestrus. Estradiol then declined in relation to the increase in LH secreation and had returned to baseline by estrus. Progesterone secretion by the corpora lutea of the cycle also increased on the afternoon of D-1 and reached a maximum value of 25-30 ng/ml early on the morning of D-2. At this time, a precipitious fall in progesterone occurred, returning to baseline values of 5-1- ng/ml by 0700 on D-2 signifying the regression of the corpora lutea of the cycle. Progesterone remained low thereafter until the afternoon of proestrus when levels increased in response to the proestrus when levels increased in response to the proestrous surge of LH. Following cervical stimulation at 1900 hr on proestrus, no differences were noted, with respect to the estrous cycle, in LH, FSH or estradiol secreation through the afternoon of D-2. Surprisingly, progesterone levels did not differ in the cycle and pseudopregnancy until the early morning of D-29 instead of progesterone levels falling to baseline as they had during the cycle, the corpora lutea of pseudopregnancy were rescused, progesterone increasing dramatically to reach levels of 45-50 ng/ml by 1700 hr on that same day. The only difference in hormone secretion that was noted which could account for this marked divergence in progesterone secretion was the pattern of prolactin secretion following cervical stimulation. In contrast to the low levels seen during the estrous cycle, biphasio surges of prolactin secretion occured each day, one being nocturnal (0100-0900 hr) and the other diurnal (1500-2100 hr). The rescue of the corpus luteum occured in association with the nocturnal surge on D-2. These results suggest that nocturnal surge on D-2, PROLACTIN IS THE MAJOR Luteotropic stimulus which transforms and estrous cycle into pseudopregnancy by prolonging progesterone secretion from the corpus luteum. Moreover, if LH is important for progesterone secretion, no changes were observed in the pattern of LH secretion which can account for the rescue of the corpus luteum.
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            Pituitary LHRH receptors during puberty in the female rat: is the proestrous decline in receptor content due to occupancy?

            As during the adult estrous cycle, the number of pituitary luteinizing hormone-releasing hormone (LHRH) receptors was found to increase prior to the first preovulatory surge of LH. Receptor content, as measured by the binding of the analog 125I-D-Ala6-Pro9-LHRH to pituitary membranes, declined markedly at the time of the first LH surge, remaining low during the first estrus and first diestrus. When pituitary membranes from animals undergoing an LH surge were incubated with MgCl2 to dissociate endogenously bound hormone, available binding sites were restored to pre-LH surge values. The increase in LHRH binding capacity was not related to an increase in receptor affinity, and it was not demonstrable in other pubertal phases. MgCl2 may have unmasked receptors made cryptic at the time of the LHRH-induced LH surge. Nevertheless, the results are compatible with the view that the proestrous decline in LHRH receptor content is, at least in part, due to occupancy of binding sites by endogenous hormone.
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              ACUTE EFFECTS OF OVARIECTOMY ON PITUITARY LH, UTERINE WEIGHT, AND VAGINAL CORNIFICATION.

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                bjb
                Brazilian Journal of Biology
                Braz. J. Biol.
                Instituto Internacional de Ecologia (São Carlos )
                1678-4375
                November 2002
                : 62
                : 4a
                : 609-614
                Affiliations
                [1 ] Universidade Estadual de Campinas Brazil
                Article
                S1519-69842002000400008
                10.1590/S1519-69842002000400008
                12659010
                60fb205c-6d29-4b3d-bdd3-d1545da303d6

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1519-6984&lng=en
                Categories
                BIOLOGY

                General life sciences
                estrous cycle,rat,cytology,smear,ciclo estral,ratas,citologia,esfregaço
                General life sciences
                estrous cycle, rat, cytology, smear, ciclo estral, ratas, citologia, esfregaço

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