Determining the Stage of the Estrous Cycle in Female Mice by Vaginal Smear

2016 ◽  
Vol 2016 (8) ◽  
pp. pdb.prot094474 ◽  
Author(s):  
Gabriel Gonzalez
2017 ◽  
Vol 35 (1) ◽  
pp. 71
Author(s):  
Siska Adelya Ramadhani ◽  
Iman Supriatna ◽  
Ni Wayan Kurniani Karja ◽  
Adi Winarto

Gosipol is a substances contained in extracted cotton seed which is thought to have the antifertility ability therefore it is often used as a herbal contraceptive. The aim of this study were to assess the folliculogenesis in mice after administrated with cottonseed extract. 60 female mice strain DDY which was 14-15 weeks old and 30-35 g body weight were divided into five groups and given cottonseed extract each 0; 1,5; 2,1 and 2,7 g/kg BW for 5, 10, 15, 24, and 24 + 10 days (without cottonseed treatment). At the end of the treatment period, mice was euthanasia to observe follicular development histomorphology (each three mice of each treatment). Mice estrous status were evaluated based on the description of the vaginal smear cells with Giemsa staining. The results showedthat the number of developing follicles was low (P < 0.05) compared with control after 5 days cottonseed extract administration at dose 2,7 g/kg BW that were 23 ± 3,6. At dose 1,5 and 2,1 g/kg BW the number of follicles was low after 24 days that were 25 ± 10,4 and 27 ± 3,5. Recovery effects of follicle number after cottonseed extract administration for 24 days was the best at a dose of 1,5 g/kg BW. Prolonge of estrous cycle occured in mice which were administrated the cottonseed extract of at all dose treatment. In conclusion, although the decrease in the number of developing follicles and prolonge of estrous cycles occurred after cottonseed extract administration, but these effects are reversible after the administration ended.


2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Naira Silva Mansano ◽  
Tabata Mariz Bohlen ◽  
Renata Frazao

Abstract It is well known that nutritional status affects the reproduction, since an adequate amount of energy is necessary for puberty onset and fertility. However, the neural mechanisms by which energy homeostasis affects reproduction is not completely elucidated. To determine if acute or chronic food restriction (FR) are able to modulate the estrous cycle, adult female mice were used in the experiments. The estrous cycle was evaluated by daily observation of vaginal smear. To determine the effects of an acute FR protocol on estrous cycle, females were individualized and kept on ad libitum diet (control, n=17) or fasted for 24 hours (n= 21). A subgroup of animals was euthanized shortly after the 24-hours test to collect hypothalamus and determinate Kiss1 mRNA levels, while another group of mice were regrouped and fed ad libitum. To determine the effects of a chronic FR protocol on estrous cycle, control mice were individualized and maintained with 100% of daily food content (average of 5 g per day, n = 6), or submitted to 60% of FR (n= 12). Animals were fed ad libitum after test. As expected, mice fasted for 24-hours exhibited a significant weight loss (control: 21.7 g ± 0.5 vs 21.6 ± 0.5 g; fasted: 22.7g ± 0.5 vs 18.7g ± 0.4, P=0.0001). This effect was followed by a significant reduction of hypothalamic Kiss1 mRNA expression (control: 1.0 ± 0.2; fasted: 0.3 ± 0.05, P=0.04, n=4/4 per group). Surprisingly, even under lower Kiss1 mRNA levels, 24-hours fasting induced no changes on estrous cycle. On the other hand, chronic FR induced a gradual weight loss (body weight at the 5th day of FR, control: 21.5g ± 0.2; FR: 17.3g ± 0.7, P=0.0002). The chronic FR was follow by the disruption of estrous cyclicity. While control mice exhibited a regular pattern of cyclicity during the period of evaluation, only leukocytes were identified in the vaginal smear of mice submitted to 60% of FR, even though they had a normal cycling pattern before the test. Therefore, by comparing 30 days of estrous cycle evaluation, including the period before chronic FR, while control mice exhibited cornified cells in the vaginal smear 58.5 ± 4.9% of days, female mice submitted to FR exhibited cornified cells in 38.3 ± 3.8% of days (P= 0.0068). Approximately 3-4 days after the end of the chronic FR females returned to exhibit estrous cyclicity, however the length of the estrous cycle was prolonged compared to control group. Our data suggest that chronic nutritional status variations are required to disrupt the hypothalamus-pituitary-gonadal axis and therefore the estrous cyclicity.


1983 ◽  
Vol 17 (2) ◽  
pp. 233-236 ◽  
Author(s):  
A. Marchlewska-Koj ◽  
M. Kruczek ◽  
E. Toch
Keyword(s):  

2002 ◽  
Vol 62 (4a) ◽  
pp. 609-614 ◽  
Author(s):  
F. K. MARCONDES ◽  
F. J. BIANCHI ◽  
A. P. TANNO

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.


2012 ◽  
Vol 226 (2) ◽  
pp. 376-380 ◽  
Author(s):  
Carolyn J. Koonce ◽  
Alicia A. Walf ◽  
Cheryl A. Frye

Author(s):  
Ologhaguo Macstephen Adienbo ◽  
Ogechi Stephanie Ezeala

Aim: Reports that some phytochemicals interfere with reproductive functions, in both humans and animals necessitated this study which is aimed at determining the effects of fruit extract of Tetrapleuratetrapteraon oestrous cycle and ovarian functions in females. Methods: Adult female wistar rats weighing 160-180 g with regular 4-5 days oestrus cycle were selected into 4 groups of 6 animals each. Group 1 (control) administered 1ml distilled water; groups II, III and IV were daily treated with the extract at doses 75 mg/kg, 150mg/kg and 300 mg/kg body weight respectively, orally for 21 days. Microscopic evaluation of vaginal smear was done daily to determine the various stages of the estrous cycle, their duration, as well as the estrous cycle length. After 24 hours of last administration, each rat was weighed, sacrificed, and right ovary was homogenised and the homogenate used for analyses of total protein content, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) concentration, according to standard protocols. Results: There was significant (P < 0.05) increase in duration of diestrus phase and estrous cycle length in all the extract-treated groups, compared to control animals. Also, there was relative reductions in the duration for proestrus (p<0.05), estrus (p<0.05) and metestrus (p<0) phases of the cycle, with a relative increase in duration for diestrus phase (p<0.05) in animals treated with 150 mg/kg and 300 mg/kg body weight respectively. In addition, a significant (P < 0.05) increase was observed in ovarian Protein, and Superoxide dismutase (SOD) enzyme activity; as well as significant (P < 0.05) reduction in malondialdehyde (MDA) level and in weight gain in the test animals, compared to the control. Conclusion:Tetrapleuratetrapterafruit extract disrupts ovarian utilisation of proteins in the ovaries, thereby impairing oestrous cyclicity, and body weight. These could result to infertility.


Author(s):  
Nazia Begum ◽  
Kandavalli Manipriya ◽  
Rahathunnisa Begum ◽  
Veeresh B

Rat estrous cycle determination or vaginal smear staining is paramount for studies related to endocrinology and reproduction; in the present study; we have reported a simple and rapid method for estrous cycle determination in rats using crystal violet. With this technique, the identification of stages can be done even on the next day. Hormonal variation in blood and histomorphological changes in ovaries at different stages of the estrous cycle were studied in female Wistar rats, which can be used to determine the hormone levels in works related to hormonal drugs, further ovarian morphology can be used to study changes in ovaries during the estrous cycle. This study aims to report a rapid and simple method for vaginal cytology using crystal violet and to report normal hormonal levels and histomorphology of ovaries in various phases of the estrous cycle.


Author(s):  
Swati B. Pokale ◽  
Ghanshayam Jadhav

This study evaluates the prospective use of an herbal plant Piper longum in letrozole induced polycystic ovary syndrome using a rat model. The study used female wistar rats, which were divided into 9 groups, each containing 6 animals. Group I (Control) daily received 1% Carboxy Methyl Cellulose (CMC)as a vehicle control. Letrozole (1mg/kg) was administered by oral route for period of 21days for induction of PCOS in group (II-IX). During experimental period, vaginal smear of all females were collected daily for the estrous cycle determination. During 28 days of letrozole administration, changes in estrous cycle of females were observed and studied. This study showed that PCOS was induced. After Letrozole treatment, 6 animals from group III-IX treated orally with, standard drugs Metformin (300mg/kg/oral route), Clomiphene citrate (100mg/kg/oral route), plant drug Piper longum L. at a concentration of 200mg/kg, 400mg/kg and 800mg/kg, drug interaction groups: Metformin + Piper longum L, 800mg/kg and Clomiphene citrate +Piper lonum L., 800mg/kg body weight and studied for consecutive estrous cycles. Vaginal smear were examined, it showed that hydro alcoholic extract of fruits of Piper longum Linn. group has potential effect on PCOS bringing estrous cycle to normalcy. Also, after Letrozole treatment ovary and reproductive weights of normal rats increased which is normalizes with plant drug treatment. Further studies of hydroalcoholic extract of fruits Piper longum Linn. need to be carried out to check other related parameters of PCOS.


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