Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells

Sugiyama, Makoto; Yasunaga, Arata; Kobayashi, Ryosuke; Fukasawa, Hanae; Hashimoto, Osamu; Kurusu, Shiro; Sasada, Hiroshi; Yoshioka, Kazuki

doi:10.1007/s00441-020-03310-w

Cited by 8 publications

(4 citation statements)

References 28 publications

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“…Several attempts have been made to develop rapid, objective methods, particularly for use in mice. These include using non-stained smear material ( 27 , 40 ), modifications to staining methods ( 41 ) and application of deep learning technology for classification of smears ( 42 ). Alternatives to vaginal cytology have been proposed based on gross examination of the vaginal opening ( 43 ), changes in skin temperature due to activation of brown adipose tissue ( 44 ) and variations in vaginal wall impedence ( 45 ).…”

Section: Introductionmentioning

confidence: 99%

Effect of Estrous Cycle on Behavior of Females in Rodent Tests of Anxiety

2021

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Anxiety disorders are more prevalent in women than in men. In women the menstrual cycle introduces another variable; indeed, some conditions e.g., premenstrual syndrome, are menstrual cycle specific. Animal models of fear and anxiety, which form the basis for research into drug treatments, have been developed almost exclusively, using males. There remains a paucity of work using females and the available literature presents a confusing picture. One confound is the estrous cycle in females, which some authors consider, but many do not. Importantly, there are no accepted standardized criteria for defining cycle phase, which is important given the rapidly changing hormonal profile during the 4-day cycle of rodents. Moreover, since many behavioral tests that involve a learning component or that consider extinction of a previously acquired association require several days to complete; the outcome may depend on the phase of the cycle on the days of training as well as on test days. In this article we consider responsiveness of females compared to males in a number of commonly used behavioral tests of anxiety and fear that were developed in male rodents. We conclude that females perform in a qualitatively similar manner to males in most tests although there may be sex and strain differences in sensitivity. Tests based on unconditioned threatening stimuli are significantly influenced by estrous cycle phase with animals displaying increased responsiveness in the late diestrus phase of the cycle (similar to the premenstrual phase in women). Tests that utilize conditioned fear paradigms, which involve a learning component appear to be less impacted by the estrous cycle although sex and cycle-related differences in responding can still be detected. Ethologically-relevant tests appear to have more translational value in females. However, even when sex differences in behavior are not detected, the same outward behavioral response may be mediated by different brain mechanisms. In order to progress basic research in the field of female psychiatry and psychopharmacology, there is a pressing need to validate and standardize experimental protocols for using female animal models of anxiety-related states.

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Section: Introductionmentioning

confidence: 99%

Effect of Estrous Cycle on Behavior of Females in Rodent Tests of Anxiety

2021

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“…östrus fazı öncesi dönemden elde edilen smear örneklerinde, vajen epitelinin yüzeyini kaplayan mukus hücrelerine odaklanarak, bu hücrelerin PAS ve Alcian blue ile boyandığını göstermişlerdir. Optimum çiftleşme zamanı için Alcian blue ve PAS ile boyalı preparatların, Giemsa boyasına göre daha doğru sonuçlar verdiğini iddia etmişlerdir (21). Spontan ovulasyonla elde edilen oositleri değerlendiren bir çalışmada, Toluidin mavisi kullanarak, smearde geniş kornifiye epitelin gözlendiği aşamada denekleri sakrifiye edip tuba uterinanın ampulla bölgesinden oosit elde ettiklerini belirtmişlerdir (13) (Şekil 3).…”

Section: öStrus Döngüsünde çEvresel Etkenlerunclassified

An Easy Way to Evaluate the Female Genital Canal in Mice and Rats: Vaginal Cytology

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Metin

2022

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Dişi genital kanalın durumunu izlemek için kullanılan vajinal sitoloji yöntemi, zaman içinde farklı metodlar ortaya çıksa da, kolay ve pratik olması nedeniyle sıklıkla tercih edilmektedir. 1917’de Stockard ve Papanicolaou’nun kobay vajinal lümen epitel hücrelerinin ovulasyon döngüsünün durumuna göre değişiklik gösterdiğini keşfetmelerinin ardından östrus siklusu takibi, dişi genital sistem çalışmalarının vazgeçilmez bir parçası olmuştur. Vajinal sitolojiyi göstermek için smear örneklerinin alınma zamanı ve yöntemi, uygulayıcının tecrübesi gibi çeşitli faktörler sonuçların doğruluğu üzerine etkilidir. Östrus siklusu, sıçan ve farelerde temel olarak dört faz gösterir: proöstrus, östrus, metöstrus ve diöstrus. Her fazın hücre tipi ve yoğunluğu farklıdır. Bu derlemede; fare ve sıçandan smear alma yöntemleri ve östrus siklus fazlarının özelliklerinin yanı sıra, elde edilen verilerin deneysel çalışmalarda kullanım alanlarına yönelik literatür bilgileri yer almaktadır. Ayrıca bu çalışma, dişi üreme sistemindeki dinamik değişikliklerin takibi, deneysel çalışmaların doğru planlanması ve sonuçların değerlendirilmesi yönünden dikkat edilmesi gereken noktaları belirtmesi ile dişi deneklerle çalışacak araştırmacılara kapsamlı bir bakış açısı sağlamayı hedeflemektedir.

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“…Estrus smears are unambiguous, whereas smears at other times are variable in cell composition (Raimondi et al, 2022). Stages other than estrus are traditionally defined by subtle, nuanced features of cytology that require a trained eye regardless of whether qualitative (Cora et al 2015;OECD, 2008;Sugiyama et al, 2021) or semi-quantitative (Hubscher et al, 2005;Paccola et al, 2013) criteria are used. Recently, machine learning tools have been developed to assist in staging cytology smears and, interestingly, these tools most readily detect the estrus stage (Sano et al, 2020;Wolcott et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

“…Stages other than estrus are traditionally defined by subtle, nuanced features of cytology that require a trained eye regardless of whether qualitative (Cora et al. 2015; OECD, 2008; Sugiyama et al., 2021) or semi‐quantitative (Hubscher et al., 2005; Paccola et al., 2013) criteria are used.…”

Section: Introductionmentioning

confidence: 99%

The Track‐by‐Day Method for Monitoring the Rodent Estrous Cycle

2023

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The exclusion of female subjects from preclinical neuroscience research has traditionally been justified in part by concerns about potential effects of cycling ovarian hormones on brain function. There is evidence that some behavioral and neurobiological measures do change over the estrous cycle and, as the use of female subjects becomes increasingly routine, there is a greater demand for accessible cycle‐tracking methods. Conventional estrous cycle staging requires expert training in the qualitative interpretation of vaginal cytology smears, which serves as a barrier for novice researchers. In addition, definitions and reporting practices are not standardized across laboratories, which makes it difficult to compare results across studies and likely contributes to a false perception of the cycle as ephemeral and inconsistent. Here, we describe a streamlined method for monitoring the estrous cycle in rats, which we term Track‐by‐Day. It is simple to implement and inherently produces consistent reporting. Our protocol should serve to demystify and facilitate adoption of cycle tracking for those new to the practice. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Collection and staining of vaginal smears Basic Protocol 2: Track‐by‐Day classification of vaginal smears Support Protocol: Preparation of gelatin‐subbed slides

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Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells

Cited by 8 publications

References 28 publications

Effect of Estrous Cycle on Behavior of Females in Rodent Tests of Anxiety

Effect of Estrous Cycle on Behavior of Females in Rodent Tests of Anxiety

An Easy Way to Evaluate the Female Genital Canal in Mice and Rats: Vaginal Cytology

The Track‐by‐Day Method for Monitoring the Rodent Estrous Cycle

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