Assessing Reproductive Status/Stages in Mice

Claudia S. Caligioni1

1 Cellular and Develomental Neurobiology, NIH/NINDS, Bethesda Maryland
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Appendix 4I
DOI:  10.1002/0471142301.nsa04is48
Online Posting Date:  July, 2009
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Abstract

The short reproductive cycle length observed in rodents, called the estrous cycle, makes them an ideal animal model for investigation of changes that occur during the reproductive cycle. Most of the data in the literature about the estrous cycle is obtained from rats because they are easily manipulated and they exhibit a clear and well‐defined estrous cycle. However, the increased number of experiments using knockout mice requires identification of their estrous cycle as well, since (in)fertility issues may arise. In mice, like rats, the identification of the stage of estrous cycle is based on the proportion of cell types observed in the vaginal secretion. The aim of this unit is to provide guidelines for quickly and accurately determining estrous cycle phases in mice.Curr. Protoc. Neurosci. 48:A.4I.1‐A.4I.8. © 2009 by John Wiley & Sons, Inc.

Keywords: estrous cycle; mouse; vaginal smears; wet vaginal

     
 
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Table of Contents

  • Introduction
  • Description of Stages of Estrous Cycle and Hormonal Considerations
  • Basic Protocol 1: Assessment of Vaginal Opening in Mice
  • Basic Protocol 2: Collection and Analyses of Mouse Vaginal Secretion (“Wet Smears”)
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Assessment of Vaginal Opening in Mice

  Materials
  • Female mice (from 23‐day‐old)
  • 0.9% saline
  • Cotton balls

Basic Protocol 2: Collection and Analyses of Mouse Vaginal Secretion (“Wet Smears”)

  Materials
  • Post‐pubertal female mice (∼40 days old)
  • Phosphate‐buffered saline (PBS; appendix 2A) or 0.9% saline (0.15 M)
  • Fine‐tipped plastic pipets, 1‐mm tip i.d.
  • Cleaned slides or printed slides
  • Light microscope with 10× objective
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Figures

Videos

Literature Cited

Literature Cited
   Caligioni, C.S. and Franci, C.R. 2002. Oxytocin secretion induced by osmotic stimulation in rats during the estrous cycle and after ovariectomy and hormone replacement therapy. Life Sci. 71:2821‐2831.
   Freeman, M.E. 1994. The neuroendocrine control of the ovarian cycle in the rat. In Physiology of Reproduction (E. Knobil and J.D. Neill, eds.) pp. 613‐658. Raven Press, New York.
   Marcondes, F.K., Bianchi, F.J., and Tanno, A.P. 2002. Determination of the estrous cycle phases of rats: Some helpful considerations. Braz. J. Biol. 62:609‐614.
   Nelson, J.F., Felicio, L.S., Randall, P.K., Sims, C., and Finch, C.E. 1982. A longitudinal study of estrous cyclicity in aging C57BL/6J mice: I. Cycle frequency, length and vaginal cytology. Biol. Reprod. 27:327‐339.
   Ojeda, S.R. and Urbancki, H.F. 1994. Puberty in the rat. In Physiology of Reproduction (E. Knobil and J.D. Neill, eds.) pp. 363‐409. Raven Press, New York.
   Parkening, T.A., Collins, T.J., and Smith, E.R. 1982. Plasma and pituitary concentrations of LH, FSH, and prolactin in aging C57BL/6 mice at various times of the estrous cycle. Neurobiol. Aging 3:31‐35.
   Rodriguez, I., Araki, K., Khatib, K., Martinou, J‐C., and Vassalli, P. 1997. Mouse vaginal opening in an apoptosis‐dependent process which can be prevented by the overexpression of Bcl2. Dev. Biol. 184:115‐121.
   Sarkar, D.K., Chiappa, S.A., Fink, G., and Sherwood, N.M. 1976. Gonadotropin‐releasing hormone surge in pro‐oestrous rats. Nature 264:461‐463.
   Walmer, D.K., Wrona, M.A., Hughes, C.L., and Nelson, K.G. 1992. Lactoferrin expression in the mouse reproductive tract during the natural estrous cycle: Correlation with circulating estradiol and progesterone. Endocrinology 131:1458‐1466.
   Yener, T., Turkkani Tunc, A., Aslan, H., Aytan, H., and Cantug Caliskan, A. 2007. Determination of oestrous cycle of the rats by direct examination: How reliable? Anat. Histol. Embryol. 36:75‐77.
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