- Email: [email protected]
The presence of secretoneurin in human synovium and synovial fluid
Neuroscience Letters 224 (1997) 139–141
The presence of secretoneurin in human synovium and synovial fluid U. Eder a, M. Hukkanen b, B. Leitner a, E. Mur c, P. Went d, R. Kirchmair a, R. Fischer-Colbrie a, J.M. Polak b, H. Winkler a,* a
Department of Pharmacology, University of Innsbruck, Peter-Mayr-Straße 1a, A-6020 Innsbruck, Austria b Department of Histochemistry, Royal Postgraduate Medical School, London W12 ONN, UK c Clinic of Internal Medicine, Anichstraße 35, A-6020 Innsbruck, Austria d Orthopaedic Clinic, Anichstraße 35, A-6020 Innsbruck, Austria Received 4 November 1996; revised version received 10 February 1997; accepted 10 February 1997
Abstract Secretoneurin is a neuropeptide formed from the proprotein secretogranin II. It is found in afferent nerve fibres and has chemotactic activity for monocytes, neutrophils and fibroblasts. We investigated the presence of secretoneurin in synovial fluid and synovium from patients with osteoarthritis and rheumatoid arthritis. The secretoneurin immunoreactive material found in synovial fluid was identified by high performance liquid chromatography as the free peptide secretoneurin. Its level in hip joints was 15.6, in knee joints of osteoarthritis patients 17.3 and in rheumatoid patients significantly lower (8.6 fmol/ml). Immunocytochemistry provided evidence for the presence of sub-intimal secretoneurin-immunoreactive nerve fibres in knee synovium in osteoarthritic patients. In rheumatoid synovium, only very few immunoreactive fibres were found these being mostly localised in deep stroma. The results show that secretoneurin is present in osteoarthritic joint and suggest that secretoneurin levels are down-regulated in rheumatoid joint. Therefore, secretoneurin may participate in acute or mild phases of inflammation but is unlikely to have a major role when more severe inflammation is present such as that seen in rheumatoid joint. 1997 Elsevier Science Ireland Ltd. Keywords: Secretogreanin II; Neuropeptide; Inflammation; Arthritis
In order to define causative factors in inflammatory diseases of the joints many studies have concentrated on neuropeptides as possible candidates. In human synovium afferent nerve fibres immunoreactive for substance P (SP) and calcitonin gene related peptide (CGRP) have been demonstrated [9,17]. These neuropeptides have also been found in the synovial fluid, however, the results appear controversial. Several studies found more than 100 fmol SP/ml [2,5,6,18,19], whereas in two other studies either no SP [16] or a very low level (,3.5 fmol/ml; [11]) was detected. It should be noted that in this latter study the immunoreactive material was characterized by high performance liquid chromatography (HPLC). It was found that immunoreactive material in unextracted samples obviously not related to SP interfered with the radioimmunoassay. In the study by Larson et al. [16], although no SP could be detected, neurokinin A (about 5 fmol/ml), CGRP (about 15 fmol/ml) and neuropeptide Y (NPY; about 4 fmol/ml) were * Corresponding author. Tel.: +43 512 5073700; fax: +43 512 5072868; e-mail: [email protected]
present in synovial fluid of controls. There is also no agreement whether these neuropeptide levels are changed in rheumatic diseases. In the careful study by Joyce et al. [11] there was no evidence that SP levels were increased in various diseases of the joint, including osteoarthritis and rheumatoid arthritis. In the other study [16] neurokinin A (NKA) was decreased in arthritic joints, whereas CGRP and NPY were increased. We have recently characterized a new neuropeptide, i.e. secretoneurin [13] which is derived from the peptide secretogranin II (see [7]). Secretoneurin can release dopamine from central neurons [1,21] and it has, which is of interest in the present context, chemotactic activity for monocytes and fibroblasts but not neutrophils [12,20,22]. Its message is present in spinal ganglia and secretoneurin is stored in and released from afferent C-fibres [14]. Since we have developed [13] a very sensitive and reliable assay for this peptide, we have analysed synovial fluid samples to provide evidence that secretoneurin is present and have also used immunocytochemistry to demonstrate its presence in synovium itself.
0304-3940/97/$17.00 1997 Elsevier Science Ireland Ltd. All rights reserved PII S0304-3940 (97 )1 3467-X
140
U. Eder et al. / Neuroscience Letters 224 (1997) 139–141
Synovial fluid samples were boiled for 12 min (secretoneurin is heat stable) and then centrifuged for 20 min. The supernatants were lyophilised and used for further analysis. The specific and sensitive (detection limit 1–4 fmol) radioimmunoassay (RIA) for secretoneurin and HPLC with molecular exclusion column have been described previously [13]. Fig. 1 demonstrates the results obtained by HPLC chromatography. The immunoreactive material eluted exactly in the position of the free peptide secretoneurin. We have previously shown [13] that in neurons the propeptide secretogranin II is completely processed to free secretoneurin. Thus afferent nerve fibres are likely to release the free peptide. Having demonstrated that our RIA specifically measured free secretoneurin we obtained quantitative data. In synovial fluid from the hip joint (obtained during operations for hip replacement) in osteoarthritic patients the secretoneurin level was 15.6 ± 2.5 (n = 4), in the knee joints of patients with osteoarthritis it was 17.3 ± 2.9 (n = 6) and in those of rheumatoid arthritis it was lower (9.5 ± 1.6, n = 7, P , 0.05). Since secretoneurin is also found in serum (20 fmol/ml, unpublished observation) one might argue that secretoneurin in synovial fluid is derived from this source. However, at least in rats no correlation between serum and synovial fluid levels of neuropeptides (SP, CGRP, and NPY) could be found [4]. It seems therefore likely that secretoneurin in the synovial fluid is derived from nerves in the synovium. In order to investigate this further, we studied the possible presence of secretoneurin-immunoreactive nerve fibres in human synovium. Samples of knee synovium were collected from patients with osteoarthritis (n = 6) or rheumatoid arthritis (n = 6). All tissues were immediately fixed
Fig. 1. Molecular size exclusion chromatography of extracts from synovial fluid. A heat-stable extract from a synovial fluid from a patient with rheumatoid arthritis patient was subjected to HPLC followed by RIA. The elution position of the free peptide secretoneurin (SN) is indicated by an arrow. Analogous results were obtained in three experiments.
Fig. 2. Secretoneurin- (A) and CGRP-immunoreactive nerve fibres (B) in synovial subintima from a patient with osteoarthritis. Secretoneurin- and CGRP-immunoreactive nerve fibres are found both around synovial vasculature and as free ending terminals (arrows) in close proximity to the synovial intima. sc, synovial cavity; fi, a mononuclear inflammatory cell infiltrate. Original magnification × 350.
after removal by immersion for 16–18 h at 4°C in freshly made Zamboni’s solution. After fixation, the tissues were washed in phosphate-buffered saline (PBS) containing 15% (w/v) sucrose and 0.01% (w/v) sodium azide at 4°C before mounting, cryosectioning and immunocytochemical processing. Antisera used included polyclonal antiserum raised against synthetic secretoneurin, SP and CGRP and antiserum for protein gene product 9.5 (PGP 9.5) which was used for visualisation of the overall synovial innervation. Avidin-biotin peroxidase method was used for immunocytochemical localization of the above peptides and proteins with glucose oxidase nickel amplification. The technique is described in detail elsewhere [10]. As reported previously, rheumatoid inflammatory synovium was found to be much less densely innervated compared with osteoarthritic synovium [9,15,17]. In osteoarthritic synovium, the distribution of secretoneurin-immunoreactive nerve fibres paralleled that seen for SP-immunoreactive fibres being less frequently found than those of CGRP- and PGP-9.5-immunoreactive fibres. These secretoneurin-immunoreactive fibres were
U. Eder et al. / Neuroscience Letters 224 (1997) 139–141
found around synovial vasculature and in parallel and terminating to the subintima of the synovial layer providing morphological evidence that secretoneurin may in fact be released in close proximity to the synovial cavity (Fig. 2). The present study has established with reliable and specific methods that the peptide secretoneurin is present in synovial nerves and is found in the synovial fluid. The concentrations are in the range of those being reported for CGRP and NKA [16]. Synovial fluid levels of secretoneurin in patients with rheumatoid arthritis are significantly lower. As discussed above Larsson et al. [16] found also a decrease for NKA in such patients, whereas CGRP and NPY were increased. For SP no changes were observed in the two studies discussed above. There was also no constant change of SP levels in synovial fluid in antigenic monoarthritis in rats [8] and only in acutely induced joint inflammation the levels of SP, NKA, CGRP and NPY were increased [3]. As shown by immunohistochemistry secretoneurin immunoreactive fibres are less frequent in rheumatoid joints than in those of osteoarthritic patients. Lack of immunostaining might be due to a loss of peptide caused by increased secretion. However, our data on the synovial fluid levels of secretoneurin which are lower in rheumatoid patients, do not support such a possibility. In conclusion our study has firmly established that the neuropeptide secretoneurin is present in synovial nerve fibres and in synovial fluid. In rheumatoid patients secretoneurin-positive fibres are less frequently observed in the synovium and its level in synovial fluid is decreased. Thus, secretoneurin may participate in acute phases of synovial inflammatory but like other neuropeptides it is unlikely to have a major role in the more prolonged inflammatory processes of the joints as occurring in rheumatoid patients. [1] Agneter, E., Sitte, H.H., Sto¨ckl-Hiesleitner, S., Fischer-Colbrie, R., Winkler, H. and Singer, E.A., Sustained dopamine release induced by secretoneurin in the striatum of the rat: a microdialysis study, J. Neurochem., 65 (1995) 622–625. [2] Agro, A. and Stanisz, A.M., Are lymphocytes a target for substance P modulation in arthritis, Semin. Arthritis Rheum., 21 (1992) 252–258. [3] Bilevicute, I., Lundeberg, T., Ekblom, A. and Theodorsson, E., Bilateral changes of substance P-, neurokinin A-, calcitonin gene-related peptide- and neuropeptide Y-like immunoreactivity in rat knee joint synovial fluid during acute monoarthritis, Neurosci. Lett., 153 (1993) 37–40. [4] Bilevicute, I., Lundeberg, T., Ekblom, A. and Theodorsson, E., Substance P-, neurokinin A-, calcitonin gene related peptide- and neuropeptide Y-like immunorectivity (-LI) in rat knee joint synovial fluid during acute monoarthritis is not correlated with concentrations of neuropeptide-LI in cerebrospinal fluid and plasma, Neurosci. Lett., 167 (1994) 145–148. [5] De Miguel, E., Arnalich, F., Tato, E., Vazquez, J., Gijon-Banos, J. and Hernanz, A., The effect of gold salts in rheumatoid arthritis, Neurosci. Lett., 174 (1994) 185–187. [6] Devillier, P., Weill, B., Renoux, M., Menkes, C. and Pradelles, P., Elevated levels of tachykinin-like immunoreactivity in joint fluids from patients with rheumatic inflammatory diseases, N. Engl. J. Med., 314 (1986) 1323.
141
[7] Fischer-Colbrie, R., Laslop, A. and Kirchmair, R., Secretogranin II: molecular properties, regulation of biosynthesis and processing to the neuropeptide secretoneurin, Prog. Neurobiol., 46 (1995) 49–70. [8] Garrett, N., Kidd, B., Cruwys, S. and Tomlinson, D., Changes in preprotachykinin mRNA expression and substance P levels in dorsal root ganglia of monoarthritic rats: comparison with changes in synovial substance P levels, Brain Res., 675 (1995) 203–207. [9] Gro¨nblad, M., Konttinen, Y.T., Korkala, O., Liesi, P., Hukkanen, M. and Polak, J.M., Neuropeptides in synovium of patients with rheumatoid arthritis and osetoarthritis, J. Rheumatol., 15 (1988) 1807– 1810. [10] Hukkanen, M., Konttinen, Y.T., Santavirta, S., Paavolainen, P., Gu, X.H., Terenghi, G. and Polak, J.M., Rapid proliferation of calcitonin gene-related peptide-immunoreactive nerves during healing of rat tibial fracture suggests neural involvement in bone growth and remodelling, Neuroscience, 54 (1993) 969–979. [11] Joyce, T., Yood, R. and Carraway, R., Quantitation of substance-P and its metabolites in plasma and synovial fluid from patients with arthritis, J. Clin. Endocrinol. Metab., 77 (1993) 632–637. [12] Ka¨hler, C.M., Bellmann, R., Reinisch, N., Schratzberger, P., Gruber, B. and Wiedermann, C.J., Stimulation of human skin fibroblast migration by the neuropeptide secretoneurin, Eur. J. Pharmacol., 304 (1996) 135–139. [13] Kirchmair, R., Hogue-Angeletti, R., Gutierrez, J., Fischer-Colbrie, R. and Winkler, H., Secretoneurin – a neuropeptide generated in brain, adrenal medulla and other endocrine tissues by proteolytic processing of secretogranin II (chromogranin C), Neuroscience, 53 (1993) 359–365. [14] Kirchmair, R., Marksteiner, J., Troger, J., Mahata, S.K., Mahata, M., Donnerer, J., Amann, R., Fischer-Colbrie, R., Winkler, H. and Saria, A., Human and rat primary C-fibre afferents store and release secretoneurin, a novel neuropeptide, Eur. J. Neurosci., 6 (1994) 861–868. [15] Konttinen, Y.T., Rees, R., Hukkanen, M., Gro¨nblad, M., Tolvanen, E., Gibson, S.J., Polak, J.M. and Brewerton, D.A., Nerves in inflammatory synovium: immunohistochemical observations on the adjuvant arthritic rat model, J. Rheumatol., 17 (1990) 1586–1591. [16] Larsson, J., Ekblom, A., Henriksson, K., Lundeberg, T. and Theodorsson, E., Concentrations of substance P, neurokinin A, calcitonin gene related peptide, neuropeptide Y and vasoactive intestinal polypeptide in synovial fluid from knee joints in patients suffering from rheumatoid arthritis, Scand. J. Rheumatol., 20 (1991) 326–335. [17] Mapp, P., Kidd, B., Gibson, S., Terry, J., Revell, P., Ibrahim, N., Blake, D. and Polak, J., Substance P-, calcitonin gene related peptide- and C-flanking peptide of neuropeptide Y-immunoreactive fibres are present in normal synovium but depleted in patients with rheumatoid arthritis, Neuroscience, 37 (1990) 143–153. [18] Marshall, W., Chiu, B. and Inman, R., Substance P and arthritis: analysis of plasma and synovial fluid levels, Arthritis Rheum., 33 (1990) 87–90. [19] Menkes, C., Renoux, M., Laoussadi, S., Mauborgne, A., Bruxelle, J. and Cesselin, F., Substance P levels in the synovium and synovial fluid from patients with rheumatoid arthritis and osteoarthritis, J. Rheumatol., 20 (1993) 714–717. [20] Reinisch, N., Kirchmair, R., Ka¨hler, C.M., Hogue-Angeletti, R., Fischer-Colbrie, R., Winkler, H. and Wiedermann, C.J., Attraction of human monocytes by the neuropeptide secretoneurin, FEBS Lett., 334 (1993) 41–44. [21] Saria, A., Troger, J., Kirchmair, R., Fischer-Colbrie, R., HogueAngeletti, R. and Winkler, H., Secretoneurin releases dopamine from rat striatal slices: a biological effect of a peptide derived from secretogranin II (chromogranin C), Neuroscience, 54 (1993) 1–4. [22] Schratzberger, P., Reinisch, N., Ka¨hler, C.M. and Wiedermann, C.J., Deactivation of chemotaxis of human neutrophils by priming with secretogranin II-derived secretoneurin, Regul. Pept., 63 (1996) 65– 71.
The presence of secretoneurin in human synovium and synovial fluid U. Eder a, M. Hukkanen b, B. Leitner a, E. Mur c, P. Went d, R. Kirchmair a, R. Fischer-Colbrie a, J.M. Polak b, H. Winkler a,* a
Department of Pharmacology, University of Innsbruck, Peter-Mayr-Straße 1a, A-6020 Innsbruck, Austria b Department of Histochemistry, Royal Postgraduate Medical School, London W12 ONN, UK c Clinic of Internal Medicine, Anichstraße 35, A-6020 Innsbruck, Austria d Orthopaedic Clinic, Anichstraße 35, A-6020 Innsbruck, Austria Received 4 November 1996; revised version received 10 February 1997; accepted 10 February 1997
Abstract Secretoneurin is a neuropeptide formed from the proprotein secretogranin II. It is found in afferent nerve fibres and has chemotactic activity for monocytes, neutrophils and fibroblasts. We investigated the presence of secretoneurin in synovial fluid and synovium from patients with osteoarthritis and rheumatoid arthritis. The secretoneurin immunoreactive material found in synovial fluid was identified by high performance liquid chromatography as the free peptide secretoneurin. Its level in hip joints was 15.6, in knee joints of osteoarthritis patients 17.3 and in rheumatoid patients significantly lower (8.6 fmol/ml). Immunocytochemistry provided evidence for the presence of sub-intimal secretoneurin-immunoreactive nerve fibres in knee synovium in osteoarthritic patients. In rheumatoid synovium, only very few immunoreactive fibres were found these being mostly localised in deep stroma. The results show that secretoneurin is present in osteoarthritic joint and suggest that secretoneurin levels are down-regulated in rheumatoid joint. Therefore, secretoneurin may participate in acute or mild phases of inflammation but is unlikely to have a major role when more severe inflammation is present such as that seen in rheumatoid joint. 1997 Elsevier Science Ireland Ltd. Keywords: Secretogreanin II; Neuropeptide; Inflammation; Arthritis
In order to define causative factors in inflammatory diseases of the joints many studies have concentrated on neuropeptides as possible candidates. In human synovium afferent nerve fibres immunoreactive for substance P (SP) and calcitonin gene related peptide (CGRP) have been demonstrated [9,17]. These neuropeptides have also been found in the synovial fluid, however, the results appear controversial. Several studies found more than 100 fmol SP/ml [2,5,6,18,19], whereas in two other studies either no SP [16] or a very low level (,3.5 fmol/ml; [11]) was detected. It should be noted that in this latter study the immunoreactive material was characterized by high performance liquid chromatography (HPLC). It was found that immunoreactive material in unextracted samples obviously not related to SP interfered with the radioimmunoassay. In the study by Larson et al. [16], although no SP could be detected, neurokinin A (about 5 fmol/ml), CGRP (about 15 fmol/ml) and neuropeptide Y (NPY; about 4 fmol/ml) were * Corresponding author. Tel.: +43 512 5073700; fax: +43 512 5072868; e-mail: [email protected]
present in synovial fluid of controls. There is also no agreement whether these neuropeptide levels are changed in rheumatic diseases. In the careful study by Joyce et al. [11] there was no evidence that SP levels were increased in various diseases of the joint, including osteoarthritis and rheumatoid arthritis. In the other study [16] neurokinin A (NKA) was decreased in arthritic joints, whereas CGRP and NPY were increased. We have recently characterized a new neuropeptide, i.e. secretoneurin [13] which is derived from the peptide secretogranin II (see [7]). Secretoneurin can release dopamine from central neurons [1,21] and it has, which is of interest in the present context, chemotactic activity for monocytes and fibroblasts but not neutrophils [12,20,22]. Its message is present in spinal ganglia and secretoneurin is stored in and released from afferent C-fibres [14]. Since we have developed [13] a very sensitive and reliable assay for this peptide, we have analysed synovial fluid samples to provide evidence that secretoneurin is present and have also used immunocytochemistry to demonstrate its presence in synovium itself.
0304-3940/97/$17.00 1997 Elsevier Science Ireland Ltd. All rights reserved PII S0304-3940 (97 )1 3467-X
140
U. Eder et al. / Neuroscience Letters 224 (1997) 139–141
Synovial fluid samples were boiled for 12 min (secretoneurin is heat stable) and then centrifuged for 20 min. The supernatants were lyophilised and used for further analysis. The specific and sensitive (detection limit 1–4 fmol) radioimmunoassay (RIA) for secretoneurin and HPLC with molecular exclusion column have been described previously [13]. Fig. 1 demonstrates the results obtained by HPLC chromatography. The immunoreactive material eluted exactly in the position of the free peptide secretoneurin. We have previously shown [13] that in neurons the propeptide secretogranin II is completely processed to free secretoneurin. Thus afferent nerve fibres are likely to release the free peptide. Having demonstrated that our RIA specifically measured free secretoneurin we obtained quantitative data. In synovial fluid from the hip joint (obtained during operations for hip replacement) in osteoarthritic patients the secretoneurin level was 15.6 ± 2.5 (n = 4), in the knee joints of patients with osteoarthritis it was 17.3 ± 2.9 (n = 6) and in those of rheumatoid arthritis it was lower (9.5 ± 1.6, n = 7, P , 0.05). Since secretoneurin is also found in serum (20 fmol/ml, unpublished observation) one might argue that secretoneurin in synovial fluid is derived from this source. However, at least in rats no correlation between serum and synovial fluid levels of neuropeptides (SP, CGRP, and NPY) could be found [4]. It seems therefore likely that secretoneurin in the synovial fluid is derived from nerves in the synovium. In order to investigate this further, we studied the possible presence of secretoneurin-immunoreactive nerve fibres in human synovium. Samples of knee synovium were collected from patients with osteoarthritis (n = 6) or rheumatoid arthritis (n = 6). All tissues were immediately fixed
Fig. 1. Molecular size exclusion chromatography of extracts from synovial fluid. A heat-stable extract from a synovial fluid from a patient with rheumatoid arthritis patient was subjected to HPLC followed by RIA. The elution position of the free peptide secretoneurin (SN) is indicated by an arrow. Analogous results were obtained in three experiments.
Fig. 2. Secretoneurin- (A) and CGRP-immunoreactive nerve fibres (B) in synovial subintima from a patient with osteoarthritis. Secretoneurin- and CGRP-immunoreactive nerve fibres are found both around synovial vasculature and as free ending terminals (arrows) in close proximity to the synovial intima. sc, synovial cavity; fi, a mononuclear inflammatory cell infiltrate. Original magnification × 350.
after removal by immersion for 16–18 h at 4°C in freshly made Zamboni’s solution. After fixation, the tissues were washed in phosphate-buffered saline (PBS) containing 15% (w/v) sucrose and 0.01% (w/v) sodium azide at 4°C before mounting, cryosectioning and immunocytochemical processing. Antisera used included polyclonal antiserum raised against synthetic secretoneurin, SP and CGRP and antiserum for protein gene product 9.5 (PGP 9.5) which was used for visualisation of the overall synovial innervation. Avidin-biotin peroxidase method was used for immunocytochemical localization of the above peptides and proteins with glucose oxidase nickel amplification. The technique is described in detail elsewhere [10]. As reported previously, rheumatoid inflammatory synovium was found to be much less densely innervated compared with osteoarthritic synovium [9,15,17]. In osteoarthritic synovium, the distribution of secretoneurin-immunoreactive nerve fibres paralleled that seen for SP-immunoreactive fibres being less frequently found than those of CGRP- and PGP-9.5-immunoreactive fibres. These secretoneurin-immunoreactive fibres were
U. Eder et al. / Neuroscience Letters 224 (1997) 139–141
found around synovial vasculature and in parallel and terminating to the subintima of the synovial layer providing morphological evidence that secretoneurin may in fact be released in close proximity to the synovial cavity (Fig. 2). The present study has established with reliable and specific methods that the peptide secretoneurin is present in synovial nerves and is found in the synovial fluid. The concentrations are in the range of those being reported for CGRP and NKA [16]. Synovial fluid levels of secretoneurin in patients with rheumatoid arthritis are significantly lower. As discussed above Larsson et al. [16] found also a decrease for NKA in such patients, whereas CGRP and NPY were increased. For SP no changes were observed in the two studies discussed above. There was also no constant change of SP levels in synovial fluid in antigenic monoarthritis in rats [8] and only in acutely induced joint inflammation the levels of SP, NKA, CGRP and NPY were increased [3]. As shown by immunohistochemistry secretoneurin immunoreactive fibres are less frequent in rheumatoid joints than in those of osteoarthritic patients. Lack of immunostaining might be due to a loss of peptide caused by increased secretion. However, our data on the synovial fluid levels of secretoneurin which are lower in rheumatoid patients, do not support such a possibility. In conclusion our study has firmly established that the neuropeptide secretoneurin is present in synovial nerve fibres and in synovial fluid. In rheumatoid patients secretoneurin-positive fibres are less frequently observed in the synovium and its level in synovial fluid is decreased. Thus, secretoneurin may participate in acute phases of synovial inflammatory but like other neuropeptides it is unlikely to have a major role in the more prolonged inflammatory processes of the joints as occurring in rheumatoid patients. [1] Agneter, E., Sitte, H.H., Sto¨ckl-Hiesleitner, S., Fischer-Colbrie, R., Winkler, H. and Singer, E.A., Sustained dopamine release induced by secretoneurin in the striatum of the rat: a microdialysis study, J. Neurochem., 65 (1995) 622–625. [2] Agro, A. and Stanisz, A.M., Are lymphocytes a target for substance P modulation in arthritis, Semin. Arthritis Rheum., 21 (1992) 252–258. [3] Bilevicute, I., Lundeberg, T., Ekblom, A. and Theodorsson, E., Bilateral changes of substance P-, neurokinin A-, calcitonin gene-related peptide- and neuropeptide Y-like immunoreactivity in rat knee joint synovial fluid during acute monoarthritis, Neurosci. Lett., 153 (1993) 37–40. [4] Bilevicute, I., Lundeberg, T., Ekblom, A. and Theodorsson, E., Substance P-, neurokinin A-, calcitonin gene related peptide- and neuropeptide Y-like immunorectivity (-LI) in rat knee joint synovial fluid during acute monoarthritis is not correlated with concentrations of neuropeptide-LI in cerebrospinal fluid and plasma, Neurosci. Lett., 167 (1994) 145–148. [5] De Miguel, E., Arnalich, F., Tato, E., Vazquez, J., Gijon-Banos, J. and Hernanz, A., The effect of gold salts in rheumatoid arthritis, Neurosci. Lett., 174 (1994) 185–187. [6] Devillier, P., Weill, B., Renoux, M., Menkes, C. and Pradelles, P., Elevated levels of tachykinin-like immunoreactivity in joint fluids from patients with rheumatic inflammatory diseases, N. Engl. J. Med., 314 (1986) 1323.
141
[7] Fischer-Colbrie, R., Laslop, A. and Kirchmair, R., Secretogranin II: molecular properties, regulation of biosynthesis and processing to the neuropeptide secretoneurin, Prog. Neurobiol., 46 (1995) 49–70. [8] Garrett, N., Kidd, B., Cruwys, S. and Tomlinson, D., Changes in preprotachykinin mRNA expression and substance P levels in dorsal root ganglia of monoarthritic rats: comparison with changes in synovial substance P levels, Brain Res., 675 (1995) 203–207. [9] Gro¨nblad, M., Konttinen, Y.T., Korkala, O., Liesi, P., Hukkanen, M. and Polak, J.M., Neuropeptides in synovium of patients with rheumatoid arthritis and osetoarthritis, J. Rheumatol., 15 (1988) 1807– 1810. [10] Hukkanen, M., Konttinen, Y.T., Santavirta, S., Paavolainen, P., Gu, X.H., Terenghi, G. and Polak, J.M., Rapid proliferation of calcitonin gene-related peptide-immunoreactive nerves during healing of rat tibial fracture suggests neural involvement in bone growth and remodelling, Neuroscience, 54 (1993) 969–979. [11] Joyce, T., Yood, R. and Carraway, R., Quantitation of substance-P and its metabolites in plasma and synovial fluid from patients with arthritis, J. Clin. Endocrinol. Metab., 77 (1993) 632–637. [12] Ka¨hler, C.M., Bellmann, R., Reinisch, N., Schratzberger, P., Gruber, B. and Wiedermann, C.J., Stimulation of human skin fibroblast migration by the neuropeptide secretoneurin, Eur. J. Pharmacol., 304 (1996) 135–139. [13] Kirchmair, R., Hogue-Angeletti, R., Gutierrez, J., Fischer-Colbrie, R. and Winkler, H., Secretoneurin – a neuropeptide generated in brain, adrenal medulla and other endocrine tissues by proteolytic processing of secretogranin II (chromogranin C), Neuroscience, 53 (1993) 359–365. [14] Kirchmair, R., Marksteiner, J., Troger, J., Mahata, S.K., Mahata, M., Donnerer, J., Amann, R., Fischer-Colbrie, R., Winkler, H. and Saria, A., Human and rat primary C-fibre afferents store and release secretoneurin, a novel neuropeptide, Eur. J. Neurosci., 6 (1994) 861–868. [15] Konttinen, Y.T., Rees, R., Hukkanen, M., Gro¨nblad, M., Tolvanen, E., Gibson, S.J., Polak, J.M. and Brewerton, D.A., Nerves in inflammatory synovium: immunohistochemical observations on the adjuvant arthritic rat model, J. Rheumatol., 17 (1990) 1586–1591. [16] Larsson, J., Ekblom, A., Henriksson, K., Lundeberg, T. and Theodorsson, E., Concentrations of substance P, neurokinin A, calcitonin gene related peptide, neuropeptide Y and vasoactive intestinal polypeptide in synovial fluid from knee joints in patients suffering from rheumatoid arthritis, Scand. J. Rheumatol., 20 (1991) 326–335. [17] Mapp, P., Kidd, B., Gibson, S., Terry, J., Revell, P., Ibrahim, N., Blake, D. and Polak, J., Substance P-, calcitonin gene related peptide- and C-flanking peptide of neuropeptide Y-immunoreactive fibres are present in normal synovium but depleted in patients with rheumatoid arthritis, Neuroscience, 37 (1990) 143–153. [18] Marshall, W., Chiu, B. and Inman, R., Substance P and arthritis: analysis of plasma and synovial fluid levels, Arthritis Rheum., 33 (1990) 87–90. [19] Menkes, C., Renoux, M., Laoussadi, S., Mauborgne, A., Bruxelle, J. and Cesselin, F., Substance P levels in the synovium and synovial fluid from patients with rheumatoid arthritis and osteoarthritis, J. Rheumatol., 20 (1993) 714–717. [20] Reinisch, N., Kirchmair, R., Ka¨hler, C.M., Hogue-Angeletti, R., Fischer-Colbrie, R., Winkler, H. and Wiedermann, C.J., Attraction of human monocytes by the neuropeptide secretoneurin, FEBS Lett., 334 (1993) 41–44. [21] Saria, A., Troger, J., Kirchmair, R., Fischer-Colbrie, R., HogueAngeletti, R. and Winkler, H., Secretoneurin releases dopamine from rat striatal slices: a biological effect of a peptide derived from secretogranin II (chromogranin C), Neuroscience, 54 (1993) 1–4. [22] Schratzberger, P., Reinisch, N., Ka¨hler, C.M. and Wiedermann, C.J., Deactivation of chemotaxis of human neutrophils by priming with secretogranin II-derived secretoneurin, Regul. Pept., 63 (1996) 65– 71.