Deep learning-based classification of the mouse estrous cycle stages.


Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
16 07 2020
Historique:
received: 04 02 2020
accepted: 23 06 2020
entrez: 18 7 2020
pubmed: 18 7 2020
medline: 8 1 2021
Statut: epublish

Résumé

There is a rapidly growing demand for female animals in preclinical animal, and thus it is necessary to determine animals' estrous cycle stages from vaginal smear cytology. However, the determination of estrous stages requires extensive training, takes a long time, and is costly; moreover, the results obtained by human examiners may not be consistent. Here, we report a machine learning model trained with 2,096 microscopic images that we named the "Stage Estimator of estrous Cycle of RodEnt using an Image-recognition Technique (SECREIT)." With the test dataset (736 images), SECREIT achieved area under the receiver-operating-characteristic curve of 0.962 or more for each estrous stage. A test using 100 images showed that SECREIT provided correct classification that was similar to that provided by two human examiners (SECREIT: 91%, Human 1: 91%, Human 2: 79%) in 11 s. The SECREIT can be a first step toward accelerating the research using female rodents.

Identifiants

pubmed: 32678183
doi: 10.1038/s41598-020-68611-0
pii: 10.1038/s41598-020-68611-0
pmc: PMC7366650
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

11714

Références

Cover, K. K., Maeng, L. Y., Lebron-Milad, K. & Milad, M. R. Mechanisms of estradiol in fear circuitry: Implications for sex differences in psychopathology. Transl. Psychiatry 4, e422 (2014).
doi: 10.1038/tp.2014.67
Nowogrodzki, A. Clinical research: Inequality in medicine. Nature 550, S18–S19 (2017).
doi: 10.1038/550S18a
DiCarlo, L. M., Vied, C. & Nowakowski, R. S. The stability of the transcriptome during the estrous cycle in four regions of the mouse brain. J. Comp. Neurol. 525, 3360–3387 (2017).
doi: 10.1002/cne.24282
Spencer-Segal, J. L. et al. Distribution of phosphorylated TrkB receptor in the mouse hippocampal formation depends on sex and estrous cycle stage. J. Neurosci. 31, 6780–6790 (2011).
doi: 10.1523/JNEUROSCI.0910-11.2011
Zenclussen, M. L., Casalis, P. A., Jensen, F., Woidacki, K. & Zenclussen, A. C. Hormonal fluctuations during the estrous cycle modulate heme oxygenase-1 expression in the uterus. Front. Endocrinol. (Lausanne) 5, 32 (2014).
doi: 10.3389/fendo.2014.00032
Scharfman, H. E., Mercurio, T. C., Goodman, J. H., Wilson, M. A. & MacLusky, N. J. Hippocampal excitability increases during the estrous cycle in the rat: A potential role for brain-derived neurotrophic factor. J. Neurosci. 23, 11641–11652 (2003).
doi: 10.1523/JNEUROSCI.23-37-11641.2003
Adams, C., Chen, X. & Moenter, S. M. Changes in GABAergic transmission to and intrinsic excitability of gonadotropin-releasing hormone (GnRH) neurons during the estrous cycle in mice. eNeuro 5, e0171 (2018).
doi: 10.1523/ENEURO.0171-18.2018
Meziane, H., Ouagazzal, A. M., Aubert, L., Wietrzych, M. & Krezel, W. Estrous cycle effects on behavior of C57BL/6J and BALB/cByJ female mice: Implications for phenotyping strategies. Genes Brain Behav. 6, 192–200 (2007).
doi: 10.1111/j.1601-183X.2006.00249.x
Milad, M. R., Igoe, S. A., Lebron-Milad, K. & Novales, J. E. Estrous cycle phase and gonadal hormones influence conditioned fear extinction. Neuroscience 164, 887–895 (2009).
doi: 10.1016/j.neuroscience.2009.09.011
Lebron-Milad, K. & Milad, M. R. Sex differences, gonadal hormones and the fear extinction network: Implications for anxiety disorders. Biol. Mood Anxiety Disord. 2, 3 (2012).
doi: 10.1186/2045-5380-2-3
National Institutes of Health Consideration of Sex as a Biological Variable in NIH-funded Research. Notice #NOT-OD-15–102 (2015). https://grants.nih.gov/grants/guide/notice-files/not-od-15-102.html . Accessed 9 June.
Byers, S. L., Wiles, M. V., Dunn, S. L. & Taft, R. A. Mouse estrous cycle identification tool and images. PLoS ONE 7, e35538 (2012).
doi: 10.1371/journal.pone.0035538
Becker, J. B. et al. Strategies and methods for research on sex differences in brain and behavior. Endocrinology 146, 1650–1673 (2005).
doi: 10.1210/en.2004-1142
Goldman, J. M., Murr, A. S. & Cooper, R. L. The rodent estrous cycle: Characterization of vaginal cytology and its utility in toxicological studies. Birth Defects Res. B Dev. Reprod. Toxicol. 80, 84–97 (2007).
doi: 10.1002/bdrb.20106
Gal, A., Lin, P. C., Barger, A. M., MacNeill, A. L. & Ko, C. Vaginal fold histology reduces the variability introduced by vaginal exfoliative cytology in the classification of mouse estrous cycle stages. Toxicol. Pathol. 42, 1212–1220 (2014).
doi: 10.1177/0192623314526321
MacDonald, J. K., Pyle, W. G., Reitz, C. J. & Howlett, S. E. Cardiac contraction, calcium transients, and myofilament calcium sensitivity fluctuate with the estrous cycle in young adult female mice. Am. J. Physiol. Heart Circ. Physiol. 306, H938–H953 (2014).
doi: 10.1152/ajpheart.00730.2013
Cora, M. C., Kooistra, L. & Travlos, G. Vaginal cytology of the laboratory rat and mouse: Review and criteria for the staging of the estrous cycle using stained vaginal smears. Toxicol. Pathol. 43, 776–793 (2015).
doi: 10.1177/0192623315570339
Hubscher, C. H., Brooks, D. L. & Johnson, J. R. A quantitative method for assessing stages of the rat estrous cycle. Biotechnol. Histochem. 80, 79–87 (2005).
doi: 10.1080/10520290500138422
Matsuda, S. et al. Sex differences in fear extinction and involvements of extracellular signal-regulated kinase (ERK). Neurobiol. Learn. Mem. 123, 117–124 (2015).
doi: 10.1016/j.nlm.2015.05.009
Esteva, A. et al. Dermatologist-level classification of skin cancer with deep neural networks. Nature 542, 115–118 (2017).
doi: 10.1038/nature21056
Rajpurkar, P. et al. CheXNet: Radiologist-Level Pneumonia Detection on Chest X-rays with Deep Learning, 1–7 (2017). https://arXiv.org/1711.05225v3 .
Ehteshami, B. B. et al. Diagnostic assessment of deep learning algorithms for detection of lymph node metastases in women with breast cancer. JAMA 318, 2199–2210 (2017).
doi: 10.1001/jama.2017.14585
Liu, Y. et al. Artificial intelligence-based breast cancer nodal metastasis detection: Insights into the black box for pathologists. Arch. Pathol. Lab. Med. 143, 859–868 (2019).
doi: 10.5858/arpa.2018-0147-OA
Simonyan, K. & Zisserman, A. Very deep convolutional networks for large-scale image recognition. International Conference on Learning Representations, 1–14 (2015). https://arXiv.org/1409.1556v6
Raghu, M., Zhang, C., Kleinberg, J. & Bengio, S. Transfusion: Understanding transfer learning for medical imaging. Adv. Neural Inf. Process. Syst. 32 (2019). https://arXiv.org/1902.07208 .
Srivastava, N., Hinton, G., Krizhevsky, A., Sutskever, I. & Salakhutdinov, R. Dropout: A simple way to prevent neural networks from overfitting. J. Mach. Learn. Res. 15, 1929–1958 (2014).
Dozat, T. Incorporating Nesterov momentum into Adam. in International Conference on Learning Representations 1–4 (2016).
Kingma, D. P. & Ba, J. Adam: A method for stochastic optimization. Machine Learning (2017). https://arXiv.org/1412.6980v9 .
Selvaraju, R. R. et al. Grad-CAM: Visual explanations from deep networks via gradient-based localization. ICCV https://doi.org/10.1007/s11263-019-01228-7 (2017).
doi: 10.1007/s11263-019-01228-7

Auteurs

Kyohei Sano (K)

Department of Cognitive Behavioral Physiology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chiba, Chiba, 260-8670, Japan.

Shingo Matsuda (S)

Department of Cognitive Behavioral Physiology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chiba, Chiba, 260-8670, Japan. smatsuda@ac.shoyaku.ac.jp.
Department of Pharmacotherapeutics, Showa Pharmaceutical University, 3-3165, Higashi-Tamagawagakuen, Machida, Tokyo, 194-8543, Japan. smatsuda@ac.shoyaku.ac.jp.
Department of Ultrastructural Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawahigashi, Kodaira, Tokyo, 187-8502, Japan. smatsuda@ac.shoyaku.ac.jp.

Suguru Tohyama (S)

Department of Pharmacotherapeutics, Showa Pharmaceutical University, 3-3165, Higashi-Tamagawagakuen, Machida, Tokyo, 194-8543, Japan.

Daisuke Komura (D)

Department of Preventive Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.

Eiji Shimizu (E)

Department of Cognitive Behavioral Physiology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chiba, Chiba, 260-8670, Japan.

Chihiro Sutoh (C)

Department of Cognitive Behavioral Physiology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chiba, Chiba, 260-8670, Japan. csutoh@graduate.chiba-u.jp.

Articles similaires

[Redispensing of expensive oral anticancer medicines: a practical application].

Lisanne N van Merendonk, Kübra Akgöl, Bastiaan Nuijen
1.00
Humans Antineoplastic Agents Administration, Oral Drug Costs Counterfeit Drugs

Smoking Cessation and Incident Cardiovascular Disease.

Jun Hwan Cho, Seung Yong Shin, Hoseob Kim et al.
1.00
Humans Male Smoking Cessation Cardiovascular Diseases Female
Humans United States Aged Cross-Sectional Studies Medicare Part C
1.00
Humans Yoga Low Back Pain Female Male

Classifications MeSH