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Journal of Histochemistry and Cytochemistry, Vol. 49, 1387-1396, November 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

An Opioid System in Connective Tissue: A Study of Achilles Tendon in the Rat

Paul W. Ackermanna, Mariana Speteab, Ingrid Nylanderc, Karolina Plojc, Mahmood Ahmedd, and Andris Kreicbergsa
a Department of Surgical Sciences, Section of Orthopedics, Karolinska Institute, Orthopedic Laboratory, Research Center, Karolinska Hospital, Stockholm, Sweden
b Department of Pharmaceutical Chemistry, Institute of Pharmacy, University of Innsbruck, Innsbruck, Austria
c Department of Pharmaceutical Biosciences, Division of Pharmacology, Uppsala, Sweden
d Section of Orthopaedics, Department of Surgery, The Aga Khan University Hospital, Karachi, Pakistan

Correspondence to: Paul W. Ackermann, Orthopedic Laboratory, Research Center M3:02, Karolinska Hospital, S-171 76, Stockholm, Sweden. E-mail: Paul.Ackermann@ks.se

The occurrence of endogenous opioids and their receptors in rat achilles tendon was analyzed by immunohistochemistry (IHC), radioimmunoassay (RIA), and in vitro binding assays. The investigation focused on four enkephalins, dynorphin B, and nociceptin/orphanin FQ. Nerve fibers immunoreactive to all enkephalins (Met-enkephalin, Leu-enkephalin, Met-enkephalin-Arg-Gly-Lys, Met-enkephalin-Arg-Phe) were consistently found in the loose connective tissue and the paratenon, whereas dynorphin B and nociceptin/orphanin FQ could not be detected. The majority of enkephalin-positive nerve fibers exhibited varicosities predominantly seen in blood vessel walls. Measurable levels of Met-enkephalin-Arg-Phe and nociceptin/orphanin FQ were found in tendon tissue using RIA, whereas dynorphin B could not be detected. In addition to the endogenous opioids identified, {delta}-opioid receptors on nerve fibers were also detected by IHC. Binding assays to characterize the opioid binding sites showed that they were specific and saturable for [3H]-naloxone (Kd 7.01 ± 0.98 nM; Bmax 23.52 ± 2.23 fmol/mg protein). Our study demonstrates the occurrence of an opioid system in rat achilles tendon, which may be assumed to be present also in other connective tissues of the locomotor apparatus. This system may prove to be a useful target for pharmacological therapy in painful and inflammatory conditions by new drugs acting selectively in the periphery. (J Histochem Cytochem 49:1387–1395, 2001)

Key Words: achilles tendon, connective tissue, rat, opioid peptides, enkephalins, peripheral nervous system, receptors, immunohistochemistry, radioimmunoassay, receptor binding


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