Monitoring Endocrine Function in Males: Using Intra‐Atrial Cannulas to Monitor Plasma Hormonal Dynamics in Toxicology Experiments

Patricia A. Fail1, Stephanie A. Anderson1

1 Research Triangle Institute, Research Triangle Park, North Carolina
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 16.5
DOI:  10.1002/0471140856.tx1605s13
Online Posting Date:  November, 2002
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Abstract

Monitoring Endocrine Function in Males: Using an Intra‐Atrial Cannula to Monitor Plasma Hormonal Dynamics in Toxicology Experiments (Patricia A. Fail and Stephanie A. Anderson, Research Triangle Institute, Research Triangle Park, North Carolina). Intra‐atrial cannulation provides assessment of endocrine change within animals. Protocols for permanent tethers used in short‐term 5‐ to 10‐day experiments and permanent vascular access used in long‐term (>10 days) experiments are presented. A protocol for blood processing is also included. Data from a longitudinal endocrine baseline assessment (LEBA) and an endocrine challenge test (ECT) a presented as well.

     
 
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Table of Contents

  • Basic Protocol 1: Intra‐Atrial Cannulation of a Male Rat with a Permanent Tether Apparatus
  • Basic Protocol 2: Intra‐Atrial Cannulation of a Male Rat with a Vascular Access Port
  • Support Protocol 1: Collecting and Processing Blood Samples from a Cannulated Male Rat
  • Commentary
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Intra‐Atrial Cannulation of a Male Rat with a Permanent Tether Apparatus

  Materials
  • Silicone adhesive
  • Heparinized saline with gentamicin: 0.9% (w/v) NaCl ( appendix 2A)/20 IU/ml heparin/0.05 mg/ml gentamicin sulfate, sterile
  • Male rat
  • Anesthetic
  • Iodide scrub
  • 70% (v/v) ethanol
  • Veterinary ophthalmic ointment (e.g., Paralube, Pharmaderm)
  • Microrenathane polyurethane tubing, 0.06‐cm (0.025‐in.) i.d., 0.1‐cm (0.04‐in.) o.d. and 0.1‐cm (0.04‐in.) i.d., 2‐cm (0.8‐in.) o.d. (Braintree Scientific)
  • Polysulfone anchor button with attached stainless steel spring tether (Instech Laboratories)
  • Swivel (Instech Laboratories)
  • Swivel‐to‐tether clamp (Instech Laboratories)
  • Ethylene oxide gas sterilizer
  • No. 22 scalpel blades, sterile
  • 1‐cc plastic tuberculin syringe with 23‐G blunt‐end needle, heparinized (i.e., coated with 1000 IU/ml liquid sodium heparin solution and allowed to dry)
  • Electric clipper
  • Toenail clipper or scissors
  • Hemostats and/or forceps, sterile
  • 4‐0 silk suture, sterile
  • 20‐G needles, sterile
  • Wound clips
  • Heating pad or heat lamp

Basic Protocol 2: Intra‐Atrial Cannulation of a Male Rat with a Vascular Access Port

  Materials
  • 22‐G noncoring Port‐a‐Cath needles (SIMS Deltec) attached to 5‐ml syringes filled with sterile 0.9% (w/v) NaCl (saline; appendix 2A)
  • Flush solution for ports: 0.9% (w/v) NaCl ( appendix 2A)/100 IU/ml heparin/0.05 mg/ml gentamicin sulfate, sterile
  • Surgical drape, sterile
  • Ruler with millimeter markings, sterile
  • Preclinical Mini‐Port (SIMS Deltec)
  • Heating pad or heat lamp
  • Additional reagents and equipment for rat surgery (see protocol 1)

Support Protocol 1: Collecting and Processing Blood Samples from a Cannulated Male Rat

  Materials
  • Male rat with an intra‐atrial cannulation with either a permanent tether apparatus (see protocol 1) or a vascular access port (see protocol 2)
  • Heparinized saline: 0.9% (w/v) NaCl ( appendix 2A)/20 IU/ml heparin, sterile
  • Rat jacket, steel spring tether, swivel, and polyurethane tubing with 1‐cc syringe attached at distal end (complete setup may be purchased through Strategic Applications or jackets may be custom made; Fig. ), for use with vascular access port only
  • Hand‐held tubing clamp
  • 1‐cc syringes with 23‐G (with 22‐G i.d.) blunt‐end needles, heparinized (i.e., coated with 1000 IU/ml liquid sodium heparin solution and allowed to dry)
  • 1.5‐ml heparinized blood collection tube
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Figures

Videos

Literature Cited

Literature Cited
   Coquelin, A. and Desjardins, C. 1982. Luteinizing hormone and testosterone secretion in young and old male mice. Am. J. Physiol. 243:E257‐E263.
   Dons, R.F. and Havlik, R. 1986. A multilayered cannula for long‐term blood sampling of unrestrained rats. Lab. Anim. Sci. 36:544‐547.
   Ellis, G.B. and Desjardins, C. 1982. Male rats secrete luteinizing hormone and testosterone episodically. Endocrinology 110:1618‐1627.
   Fail, P.A., Sauls, H.R., Pearce, S.W., Anderson, S.A., and Izard, M.K. 1992. Measures of pituitary and testicular function evaluated with an endocrine challenge test (ECT) in cannulated male rats. Toxicologist 12:436 (abstr. 1725).
   Fail, P.A., Pearce, S.W., Anderson, S.A., Tyl, R.W., and Gray, L.E. Jr. 1995. Endocrine and reproductive toxicity of vinclozolin (VIN) in male Long‐Evans hooded rats. Toxicologist 15:293 (abstr. 1570).
   Fail, P.A., Chapin, R.E., Price, C.J., and Heindel, J.J. 1998. General, reproductive, developmental, and endocrine toxicity of boric acid and inorganic boron‐containing compounds: A review. Reprod. Toxicol. 12:1‐18.
   Fail, P.A., Anderson, S.A., and Friedman, M.A. 1999. Response of the pituitary and thyroid to tropic hormones in Sprague‐Dawley versus Fischer 344 male rats. Toxicol. Sci. 52:107‐121.
   Harms, P.G. and Ojeda, S.R. 1974. A rapid and simple procedure for chronic cannulation of the rat jugular vein. J. Appl. Physiol. 36:391‐392.
   MacLeod, J.N. and Shapiro, B.H. 1988. Repetitive blood sampling in unrestrained and unstressed mice using a chronic indwelling right atrial catheterization apparatus. Lab. Anim. Sci. 38:603‐608.
   McKenna, M.L. and Bier, J.G. 1984. Multilayer cannula for long‐term infusion of unrestrained rats. Lab. Anim. Sci. 34:308‐310.
   O'Farrell, L., Griffith, J.W., and Lang, C.M. 1996. Histological development of the sheath that forms around long‐term implanted central venous catheters. J. Parenter. Enteral Nutr. 20:156‐158.
   Paulose, C.S. and Dakshinamurti, K. 1987. Chronic catheterization using vascular‐access‐port in rats: Blood sampling with minimal stress for plasma catecholamine determination. J. Neurosci. Methods 22:141‐146.
   Simard, J., Luthy, L., Guay, J., Belanger, A., and Labrie, F. 1986. Characteristics of interaction of the antiandrogen flutamide with the androgen receptor in various target tissues. Mol. Cell. Endocrinol. 44:261‐270.
   Suzuki, K., Koizumi, N., Hirose, H., Hokao, R., Takemura, N., and Motoyoshi, S. 1997. Blood sampling technique for measurement of plasma arginine vasopressin concentration in conscious and unrestrained rats. Lab. Anim. Sci. 47:190‐193.
   Tyl, R.W. 2002. In vivo models for male reproductive toxicity. In Current Protocols in Toxicology (M. Maines, L.G. Costa, E. Hodgson, and D.J. Reed, eds.) pp. 16.1.1‐16.1.15. John Wiley & Sons, New York.
   Viguier‐Martinez, M.C., Hochereau de Reviers, M.T., Barenton, B., and Perreau, C. 1983a. Effect of a nonsteroidal antiandrogen, flutamide, on the hypothalamo‐pituitary axis, genital tract and testis in immature rats: Endocrinological and histological data. Acta Endocrinol. 102:299‐306.
   Viguier‐Martinez, M.C., Hochereau de Reviers, M.T., Barenton, B., and Perreau, C. 1983b. Endocrinological and histological changes induced by flutamide treatment on the hypothalamo‐hypophyseal testicular axis of the adult male rat and their incidences on fertility. Acta Endocrinol. 104:246‐252.
   Weeks, J.R. and Davis, J.D. 1964. Chronic intravenous cannulas for rats. J. Appl. Physiol. 19:540‐541.
Key Reference
   Anonymous 1993. Preclinical Mini‐Port Implantable Vascular Access Device, pamphlet 40‐2931‐01A. Pharmacia‐Deltec (now SIMS Deltec), St. Paul, Minnesota.
  This pamphlet is the basis for the surgical implantation and the use of the vascular access port.
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