Inhibitory effect of amiloride and gadolinium on fine afferent nerves in the rat knee: evidence of mechanogated ion channels in joints.

Synovial joints are complex sensory organs which provide continuous feedback regarding position sense and degree of limb movement. The transduction mechanisms which convert mechanical forces acting on the joint into an electrochemical signal which can then be transmitted to the central nervous system are not well understood. The present investigation examined the effect of the mechanogated ion channel blockers amiloride and gadolinium on knee joint mechanosensitivity. In deeply anaesthetised rats (sodium thiopental: 100-120 mg/kg, i.p.), single unit extracellular recordings were made from knee joint group III (Adelta) and group IV (C) primary afferents in response to mechanical rotation of the joint. Afferent firing rate was measured before and after topical application of either amiloride (0.1 mM, 1 mM) or gadolinium (250 microM) onto the receptive field of the sensory unit and recording was continued every 10 min up to a total of 50 min. With normal rotation of the knee, joint mechanosensitivity was significantly reduced by both amiloride (P<0.0001; n=10-21) and gadolinium (P=0.001; n=12) and this effect was sustained throughout the recording period. This investigation provides the first in vivo electrophysiological evidence that joint mechanotransduction involves the activation of amiloride and gadolinium-sensitive mechanogated ion channels. Future studies to determine the mechanogated ion channel subtypes present in joints and the modulation of their gating properties during inflammation may yield novel approaches for the control of arthritis pain.

Voltage-gated calcium channels may be involved in the regulation of the mechanosensitivity of slowly conducting knee joint afferents in rat.

Voltage-gated Ca(2+) channels play an important role in the central processing of nociceptive information. Recently, it has been shown that L- and N-type voltage-gated Ca(2+) channels are also present on peptidergic, fine afferent nerve fibers in the knee joint capsule. Therefore, the influence of specific blockers for L-type (verapamil) or N-type (omega-conotoxin GVIA) Ca(2+) channels on the mechanosensitivity of slowly conducting afferents was tested in the rat knee joint. Topical application of 100 microM verapamil onto the receptive field reduced the mean response to knee joint rotation to 67+/-8% (SEM, n=12), obtained by outward rotations with a torque of 10 mNm above the mechanical threshold and compared with control movements. In the presence of 50 microM omega-conotoxin GVIA, the mean response decreased to 44+/-5% ( n=12), a reduction that was also observed during rotations of other intensities. Simultaneous application of both substances further reduced the response to 25+/-11% ( n=6). In additional experiments it was shown that L- and N-type voltage-gated Ca(2+) channels do not influence activity-dependent changes of the mechanical excitability. In conclusion, the data of the present study indicate that voltage-gated Ca(2+) channels may also be involved in the regulation of the mechanosensitivity of nociceptive nerve fiber endings.

Inhibitory effect of somatostatin on the mechanosensitivity of articular afferents in normal and inflamed knee joints of the rat.

The effect of somatostatin on the sensory activity of primary afferents was studied in normal and acutely inflamed rat knee joints. Fine afferent nerve fibers with conduction velocities of 0.9-18.0 m/s were recorded as single units. All nerve fibers tested responded to local mechanical stimulation, movements of the joint and i.a. injections of KCl (10(-4) mol, 0.1 ml) close to the joint. Somatostatin (10(-4) mol, 0.2 ml) caused no direct response of the units. In normal joints, somatostatin did not change the discharges evoked by non-noxious movements but decreased the responses to noxious movements significantly to about 63% of the responses before the application. In acutely inflamed joints, somatostatin reduced the discharges of non-noxious and of noxious movements to about 55% and 52%, respectively. Injections of somatostatin with lower concentrations (10(-6) mol, 10(-8) mol) i.a. close to inflamed joints revealed shorter and less pronounced reductions of the responses to noxious movements. In a proportion of afferents, substance P (10(-4) mol) and bradykinin (10(-4) mol) were able to increase these responses again. These data indicate that the mechanosensitivity of articular afferents in normal joints may also be regulated by several neuropeptides based on a balance of pro-inflammatory peptides such as substance P, and anti-inflammatory peptides such as somatostatin. In an inflamed joint, pro-inflammatory peptides seem to predominate resulting in a sensitization of the peripheral nerve fibers. In this case, an application of somatostatin or its analogues could be used clinically to compensate this effect.