Sexual and Reproductive Behaviors

Cheryl L. Sisk1, Leslie R. Meek2

1 Michigan State University, East Lansing, Michigan, 2 University of Minnesota at Morris, Morris, Minnesota
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 8.2
DOI:  10.1002/0471142301.ns0802s00
Online Posting Date:  May, 2001
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Abstract

The procedures described in this unit include testing procedures for male and female reproductive behaviors, gonadectomy, and hormonal treatments appropriate for inducing male and female reproductive behaviors. Because reproductive behaviors are social behaviors, and therefore require the presence of stimulus animals, the protocols in this unit also provide information on the preparation of stimulus animals. The protocols are written for use with laboratory rats, although a discussion of issues related to species differences in the study of reproductive behaviors is included.

     
 
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Table of Contents

  • Basic Protocol 1: Assessment of Mounts, Intromissions, and Ejaculations in Male Rats
  • Basic Protocol 2: Assessment of Lordosis in Female Rats
  • Support Protocol 1: Castration
  • Support Protocol 2: Ovariectomy
  • Support Protocol 3: Testosterone Treatment Using Subcutaneous Implant
  • Support Protocol 4: Testosterone Treatment by Subcutaneous Injection
  • Support Protocol 5: Estradiol and Progesterone Treatment
  • Reagents and Solutions
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Assessment of Mounts, Intromissions, and Ejaculations in Male Rats

  Materials
  • Female rats 60 to 120 days old, either ovariectomized and steroid primed (see Support Protocols protocol 42 and protocol 75) or gonad intact, one per test
  • 70% ethanol
  • Test males, either castrated and steroid primed (see Support Protocols protocol 31, protocol 53, and protocol 64) or gonad intact
  • 20‐gallon glass aquarium mounted on an open framework with tilted mirror positioned underneath for visualization of ventral side of the animal
  • Lamp(s) with dim red light bulb, if testing during dark portion of the light/dark cycle
  • Video camera, videotapes, VCR, and monitor
  • Pen and data pad or computer and appropriate software for data entry and analysis
  • Stopwatch
  • Additional reagents and equipment for estradiol and progesterone treatment if using ovariectomized rats (see protocol 7)

Basic Protocol 2: Assessment of Lordosis in Female Rats

  Materials
  • Stimulus male 60 to 120 days old, either castrated and steroid primed (see Support Protocols protocol 31, protocol 53 and protocol 64) or gonad intact; one per test
  • 70% ethanol
  • Test females, either ovariectomized and steroid primed (see Support Protocols protocol 42 and protocol 75), or gonad intact
  • 20‐gallon glass aquarium
  • Video camera, videotapes, VCR, and monitor
  • Lamp(s) with dim red light bulb, if testing during dark portion of the light/dark cycle
  • Additional reagents and equipment for testosterone treatment (see Support Protocols protocol 53 and protocol 64)

Support Protocol 1: Castration

  Materials
  • Male rat
  • Anesthetic (as recommended by IACUC)
  • Topical anti‐infective: 5% Nolvasan solution or Betadine
  • Topical antiseptic cream, antibiotic, and analgesic (if recommended by IACUC)
  • Surgical gloves, mask, and lab coat
  • Padded bench paper or surgical pads
  • Heating pad
  • Shaver
  • Surgical scissors, operating and iris
  • Mouse‐tooth forceps
  • Gauze
  • 3‐0 surgical silk and suturing needles or 9‐mm wound clips and applicator
  • Holding cage lined with paper towels

Support Protocol 2: Ovariectomy

  Materials
  • Female rat
  • Anesthetic (as recommended in IACUC guidelines)
  • Topical anti‐infective: 5% Nolvasan solution or Betadine
  • Antiseptic cream, analgesic, and antibiotic (if recommended by IACUC)
  • Surgical gloves, mask, and lab coat
  • Padded bench paper or surgical pads
  • Heating pad
  • Shaver
  • Surgical scissors, operating and iris
  • Mouse‐tooth forceps
  • Hemostats
  • Gauze
  • 3‐0 surgical silk and suturing needles or 9‐mm wound clips and applicator

Support Protocol 3: Testosterone Treatment Using Subcutaneous Implant

  Materials
  • Silicone type A medical adhesive (Factor II)
  • Crystalline testosterone (Sigma or Steraloids)
  • 70% ethanol
  • Saline solution: 0.9% NaCl
  • Castrated male rat (see protocol 3)
  • Anesthetic (e.g., methoxyflurane)
  • Topical anti‐infective: 5% Nolvasan solution or Betadine
  • Silastic medical‐grade tubing (Dow Corning 601‐285, 1.57‐mm i.d., 3.18‐mm o.d.)
  • Two 1‐ml syringes and one blunt‐ended 16‐G needle
  • Padded bench paper or surgical pads
  • Surgical gloves, mask, and lab coat
  • Shaver
  • Gauze
  • Surgical scissors
  • Forceps
  • 9‐mm wound clips and applicator

Support Protocol 4: Testosterone Treatment by Subcutaneous Injection

  • Testosterone propionate in sesame oil (see recipe for steroid hormones)
  • Surgical gloves, mask, and lab coat
  • 1‐ml syringe with 26‐G needle

Support Protocol 5: Estradiol and Progesterone Treatment

  Materials
  • β‐estradiol 3‐benzoate in sesame oil (see recipe for steroid hormones)
  • Progesterone in sesame oil (see recipe for steroid hormones)
  • Ovariectomized female rat (see protocol 4)
  • Surgical gloves, mask, and lab coat
  • Two 1‐ml syringes and 26‐G needles
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Figures

Videos

Literature Cited

Literature Cited
   Bronson, F.H. and Heideman, P.D. 1994. Seasonal regulation of reproduction in mammals. In The Physiology of Reproduction, Vol. 2 (E. Knobil and J.D. Neill, eds.) pp. 541‐611. Raven Press, New York.
   Claro, F., Del Abril, A., Segovia, S., and Guillamon, A. 1990. SBR: A computer program to record and analyze sexual behavior in rodents. Physiol. Behav. 58:589‐593.
   Dewsbury, D.A. 1979. Factor analysis of measure of copulatory behavior in three species of muroid rodents. J. Comp. Physiol.Psychol. 93:868‐878.
   Erskine, M.S. 1989. Solicitation behavior in the estrous female rat: A review. Hormones Behav. 23:573‐502.
   Erskine, M.S. 1995. Prolactin release after mating and genitosensory stimulation in females. Endocrine Rev. 16:508‐528.
   Everitt, B.J. 1990. Sexual motivation: A neural and behavioural analysis of the mechanisms underlying appetitive and copulatory responses of male rats. Neurosci. Biobehav. Rev. 15:217‐232.
   Freeman, M.E. 1994. The neuroendocrine control of the ovarian cycle of the rat. In The Physiology of Reproduction, Vol. 2 (E. Knobil and J.D. Neill, eds.) pp. 613‐658. Raven Press, New York.
   Holmes, G.M., Holmes, D.G., and Sachs, B.D. 1988. An IBM‐PC based data collection system for recording rodent sexual behavior and for general event recording. Physiol. Behav. 55:825‐828.
   Mallick, H.N., Tilakaratna, P., Manchanda, S.K., and Kumar, V.M. 1993. A computer program for recording male sex behaviour in rats. Indian J. Physiol. Pharmacol. 37:151‐156.
   McClintock, M.K. 1985. Group mating in the domestic rat as a context for sexual selection. Consequences for the analysis of sexual behavior and neuroendocrine responses. Adv. Study Behav. 15:1‐50.
   McClintock, M.K. and Adler, N.T. 1978. Induction of persistent estrus by airborne chemical communication among female rats. Horm. Behav. 11:515‐518.
   McClintock, M.K. and Anisko, J.J. 1982. Group mating among Norway rats. 1. Sex differences in the pattern and neuroendocrine consequences of copulation. Anim. Behav. 30:398‐509.
   McClintock, M.K., Anisko, J.J., and Adler, N.T. 1982. Group mating among Norway rats. 2. The social dynamics of copulation: Competition, cooperation and mate choice. Anim. Behav. 30:510‐525.
   Meisel, R.L. and Sachs, B.D. 1994. The physiology of male sexual behavior. In The Physiology of Reproduction, Vol. 2 (E. Knobil and J.D. Neill, eds.) pp. 3‐105. Raven Press, New York.
   Pfaus, J.G., Mendelson, S.D., and Phillips, A.G. 1990. A correlational and factor analysis of anticipatory and consummatory measures of sexual behavior in the male rat. Psychoneuroendocrinology 15:329‐350.
   Rakerd, B., Brigham, D.A., and Clemens, L.G. 1985. A microcomputer‐based system for recording and analyzing behavioral data regarding the sexual activity of male rodents. Physiol. Behav. 35:999‐1001.
   Sachs, B.D. and Barfield, R.J. 1976. Functional analysis of masculine copulatory behaviour in the rat. In Advances in the Study of Behaviour (J.S. Rosenblatt, R.A. Hinde, E. Shaw, and C. Beer, eds.) pp. 91‐155. Academic Press, Orlando, Fla.
   Smith, E.R., Damassa, D.A., and Davidson, J.M. 1977. Hormone administration: Peripheral and intracranial implants. Methods Psychobiol. 3:259‐279.
   Turek, F.W. and Van Cauter, E. 1994. Rhythms in reproduction. In The Physiology of Reproduction, Vol. 2 (E. Knobil and J.D. Neill, eds.) pp. 487‐540. Raven Press, New York.
   Weed, J.L. and Boone, J.L. 1992. A Macintosh computer system for collecting and analyzing rodent sexual behavior. Physiol. Behav. 52:183‐185.
Key References
   Cherry, J.A. 1993. Measurement of sexual behavior: Controls for variables. Methods Neurosci. 15:3‐15.
  Discusses several variables that influence expression of male reproductive behavior during behavioral tests.
   Meisel and Sachs, 1994. See above.
  Comprehensive review of description, measurement, analysis, and neuroendocrine control of male reproductive behavior.
   Pfaff, D.W., Schwartz‐Giblin, S., McCarthy, M.M., and Kow, L.‐M. 1995. Cellular and molecular mechanisms of female reproductive behaviors. In The Physiology of Reproduction, Vol. 2 (E. Knobil and J.D. Neill, eds.) pp. 107‐220. Raven Press, New York.
  Comprehensive review of neuroendocrine and molecular mechanisms regulating lordosis.
   Price, E.O. 1993. Practical considerations in the measurement of sexual behavior. Methods Neurosci. 15:16‐31.
  Discusses aspects of test and stimulus animals and testing environment that influence expression of reproductive behavior during behavioral tests.
   Smith, et al. 1977. See above.
  Describes methods of delivering steroid hormones to animals.
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