Substance P Content Research Articles

Interaction of substance P with epidermal growth factor and fibroblast growth factor in cyclooxygenase-dependent proliferation of human skin fibroblasts.

Substance P (SP), fibroblast growth factor (FGF), and epidermal growth factor (EGF) are mitogens for fibroblasts. EGF acts as a progression factor, whereas FGF and SP have competence factor activity. The ability of eicosanoids to regulate proliferation of fibroblasts and the increased production of prostaglandins by fibroblasts in response to the growth factors, led us to investigate the involvement of cyclooxygenase-dependent arachidonic acid metabolites in the mitogenic response of serum-starved human skin fibroblasts to SP, FGF, and EGF. We tested the interaction of a submaximal concentration of SP(10(-9)M) with baFGF(40 micrograms/ml) and EGF(0.01 microgram/ml) both on fibroblast proliferation and release of arachidonic acid metabolites. A combination of SP and EGF synergistically stimulated fibroblast proliferation and prostaglandin E2 release, whereas addition of SP to FGF-containing cultures did not affect cell growth. Inhibition of cyclooxygenase by acetylsalicylic acid augmented the growth response of fibroblasts to all: SP, FGF, and EGF. In the presence of acetylsalicylic acid, SP combined with FGF enhanced fibroblasts proliferation, whereas a combination with EGF inhibited cellular growth with respect to growth induced by EGF alone. Thus, interactions of SP with FGF and EGF differently affected the mitogenic response depending on the formation of arachidonic acid metabolites. The findings indicate that eicosanoids may be important mediators of competence and progression factor activities that may determine the effects of substance P on fibroblast proliferation in a cytokine network.

Effects of streptozotocin-induced diabetes and insulin treatment on substance P of the rat arterial wall

Substance P (SP) is present in perivascular nerves throughout the mammalian vasculature. Reports of diminished SP levels in nerve and gastrointestinal tissues of diabetic rats led us to examine SP-like immunoreactivity (SP-LI) in large arteries by RIA. Six weeks after inducing diabetes with streptozotocin (STZ), SP-LI was measured in the thoracic aorta, abdominal aorta, and the proximal superior mesenteric artery. In diabetics we measured a doubling (P < 0.01) of SP-LI in all three artery wall preparations. This finding was verified in a second experiment which included a subset of diabetics treated daily with insulin for the sixth week of the holding period. Again, we measured a two-fold or greater increase of SP-LI (P < 0.01–0.05) in arteries from the diabetics and found that insulin treatment significantly reduced SP-LI (P < 0.05). In contrast to reports of diminished SP content in other tissues of diabetic rats, our findings demonstrate that the artery wall experiences at least a two-fold increase of SP-LI in the diabetic state. Furthermore, this elevation of SP-LI is reduced by insulin. We speculate that these changes of arterial wall SP-LI may contribute to altered regulation of the vascular system in the diabetic state.

RETRACTED: A model for experimentally induced temperomandibular joint arthritis in rats: effects of carrageenan on neuropeptide-like immunoreactivity

Substance P(SP)-, neurokinin A (NKA)-, calcitonin gene-related peptide (CGRP)- and neuropeptide Y (NPY)-like immunoreactivity (-LI) was studied in rats' cerebrospinal fluid (CSF), plasma and perfusates (PF) from the temporomandibular joints (TMJs) at 2, 6 and 24 h following 0.01 ml injection of 2% carrageenan (CAR) into the right TMJ. SP-, NKA-, CGRP- and NPY-LI were significantly increased in both TMJ perfusates of the treated groups compared to controls. Generally an injection with CAR into the right TMJ induced a similar influence of the concentration of SP-, NKA-, CGRP- and NPY-LI in the CSF, plasma and PF at 2, 6 and 24 h following injection. However, the most pronounced changes in neuropeptide-LI occurred intra-articularly in the joint fluid, which indicates that both the sensory and sympathetic nervous system are activated in this joint following osteoarthritis induction by carrageenan.

Beta-endorphin, but not oxytocin, substance P or vasoactive-intestinal polypeptide, contributes to progesterone-induced prolactin secretion in monkeys.

Progesterone (P) stimulates prolactin secretion through a neural mechanism in estrogen (E)-primed female monkeys. Several peptides, including beta-endorphin (BE), oxytocin (OT), substance P (SP) and vasoactive intestinal polypeptide (VIP) are potential prolactin stimulatory factors and could mediate the effect of P. We hypothesized that the antagonism of a pivotal peptidergic neural system would block P-induced prolactin secretion and that the function of a pivotal peptidergic system would be altered by changes in gonadal steroid concentrations. Therefore it was of interest (1) to examine the effect of infusion of antagonists to these peptides on P-induced prolactin secretion, and (2) to determine BE, OT, SP and VIP levels in the hypothalamus of monkeys of various reproductive states. For the antagonist studies, female monkeys (n = 8) were spayed, adapted to a vest and tether remote sampling system and catheterized prior to antagonist challenges. E-primed monkeys received P injections 48 h prior to antagonist administration. Prolactin increased within 36-48 h of P injection. All antagonist challenges were administered in varying doses during the P-induced prolactin elevation and blood samples were collected every 10 min for prolactin determinations. The opiate antagonist, naloxone (n = 5), reduced serum prolactin in a dose-related manner with a mean IC50 of 1.5 +/- 0.6 micrograms/kg/min. The OT (n = 4), SP (n = 4) or VIP (n = 4) antagonists did not reduce serum prolactin in a dose-related manner. We previously reported that the hypothalamic content of OT is increased by ovarian hormones. To determine whether the hypothalamic content of BE, SP or VIP was related to gonadal status, the peptide levels in 4 hypothalamic regions of monkeys in various physiological states were measured. BE (ng/mg protein) in the medial basal hypothalamus (MBH) was significantly greater in adult females (17.7 +/- 6.9; n = 6) as compared to spayed females (0.6 +/- 0.2; n = 3) and juvenile females (1.8 +/- 1.1; n = 3). Hypothalamic content of SP in the preoptic area and mammillary bodies, but not the MBH, was significantly greater in gonadal intact females than spayed females. VIP content (pg/mg protein) was not significantly different between adult, spayed and juvenile females nor between adult and juvenile males in any hypothalamic area. Taken together these results support a pivotal role for BE in the neural regulation of P-induced prolactin secretion. The involvement of OT, SP, and VIP in a specific manner at the pituitary level is not indicated.

嘔吐におけるSubstance Pの役割タキキニン拮抗薬は新しい鎮吐薬になりうるか?

In order to elucidate the role of substance P (SP) in emetic responses, effects of resiniferatoxin (RTX), an ultra-potent capsaicin analogue that releases SP, were investigated in Suncus murinus. RTX induced dose-dependent emesis, and beside vomiting, flat down, increase of respiration rate, tremor and eye twitch were notable behavioral changes at a dose of 100 μg/kg. These effects were transient, and animals became normal 30 to 40 min after the injection. RTX-induced emesis was blocked by 30 mg/kg CP-99, 994 and 10 mg/kg ruthenium red. RTX also showed anti-emetic effects. Animals treated with RTX did not respond to cisplatin, copper sulfate, nicotine nor motion after one hour. The anti-emetic effects of RTX were also inhibited by ruthenium red but not by CP-99, 994. Both emetic and anti-emetic effects of RTX were apparently desensitized. Neonatal treatment with capsaicin completely prevented emetic response to RTX but not to cisplatin, copper sulfate, nicotine and motion. RTX significantly decreased the content of SP in the nucleus tractus spinalis nerve trigemini, the dorsal spinal cord but not in the dorsal vagal complex that is considered as part of emetic pattern generator. These results suggest that SP is involved in emetic responses but its action is complicated.

Effects of substance P and its antagonist spantide on corticosterone secretion and cytosolic free calcium concentration of dispersed zona fasciculata-reticularis cells of the rat adrenal cortex

Substance P (SP) did not change basal corticosterone (B) secretion of dispersed zona fasciculata-reticularis cells of the rat adrenal cortex. Conversely, spantide II (SPA), an antagonist of SP receptors, at a concentration 10(-7)/10(-6) M markedly raised it, and the effect was annulled by equimolar concentrations of SP. Both SP and SPA (10(-6) M) increased cytosolic free calcium concentration in our cell preparations; however, the response to SP was immediate, while that to SPA showed a lag-period of 4-5 min. SP concentration-dependently (from 10(-8) M to 10(-5) M) partially inhibited maximally ACTH (10(-8) M)-induced stimulation of B secretion of dispersed cells, and unexpectedly a similar effect was observed after SPA exposure. In light of these findings, the conclusion is drawn that SP, under basal conditions, does not exert a direct modulatory action of B secretion of rat adrenocortical cells. However, the possibility remains to be explored that SP may play a role in quenching, via a receptor-independent mechanism, the exceedingly high glucocorticoid responses to ACTH of rat adrenocortical cells.

Neutral endopeptidase (NEP) and its role in pathological pulmonary change with inhalation exposure to JP-8 jet fuel.

Through a simulated flightline exposure protocol, Fischer 344 rats (F344) were subjected to an aerosol/vapor mix of the military jet fuel, JP-8. Previous studies with this model of lung injury have revealed significant increases in pulmonary resistance, increased alveolar clearance of 99mTcDTPA, and a decrease in bronchoalveolar lavage fluid (BALF) concentration of the neuropeptide substance P (SP). Exposures to JP-8 were nose-only and for one hour daily. Six groups of Fischer 344 rats were exposed for 7, 28, or 56 days at two JP-8 concentrations (low dose = 469-520 mg/m3/hr, high dose = 814-1263 mg/m3/hr). Exposed groups were matched with longitudinal controls. In response to JP-8 inhalation, exposure animals demonstrated a dose-dependent as well as duration-determined reduction in BALF SP concentration. Both JP-8 concentrations caused significant pathological changes in lower pulmonary structures.

Substance P Induces Tumor Necrosis Factor in anex VivoModel System

Many studies have shown that tumor necrosis factor (TNF) is a potent soluble mediator of immunoregulation and inflammation. Neuropeptide substance P (SP) has been known to exert significant influence on production of certain inflammatory cytokine by immune cells. Immunopathogenic mechanism underlying the effect of neuropeptide substance P (SP) and the specific amino acid sequence of SP that induces TNF has not been clearly studied. Employingex vivoandin vitromodel systems, we investigated the direct effect of different sequences of SP on TNF secretion by whole blood and separated total mononuclear cells. Aliquots of blood samples (1 ml) or Ficoll–Hypaque-separated total mononuclear cells (1 × 106/ml) were cultured with different concentrations of SP and its sequences (SP 1–4, SP 4–11) or vasoactive intestinal peptide (VIP) for 24 hr at 37°C. Plasma samples and culture supernatants were assayed for TNF levels in a bioassay using a TNF-sensitive WEHI 164 subclone 13 cell line. Plasma from blood samples or lymphocytes treated with whole SP and SP 4–11 at 10−7, 10−8, and 10−9Mconcentrations induced significant production of TNF compared to negligible levels of TNF produced by SP 1–4-treated and untreated cultures. VIP at all concentrations tested did not induce TNF production and was similar to untreated control cultures. Separated mononuclear cells also produced significant levels of TNF in response to SP and SP 4–11. Anti-TNF-α antibodies neutralized the TNF induced by SP 4–11 in plasma. These studies suggest that anex vivosystem using whole blood may be an ideal model to study the effects of SP on TNF production. These studies also demonstrated that the TNF inducing activity of SP resides in the region containing amino acids 4 to 11.

Participation of presynaptic noradrenergic fibers in the suppression of alpha 2-adrenoceptor activity by substance P at the nucleus reticularis gigantocellularis of the rat.

We applied reverse microdialysis and high performance liquid chromatography (HPLC) analysis to evaluate the participation of presynaptic noradrenergic neurotransmission in the suppression by substance P (SP) of alpha 2-adrenoceptor activity at the nucleus reticularis gigantocellularis (NRGC) in Sprague-Dawley rats anesthetized with pentobarbital sodium. Microinfusion of SP (600 microM/min) into the NRGC through a stereotaxically positioned microdialysis probe attenuated the hypotensive and bradycardiac actions of the alpha 2-adrenoceptor agonist, guanabenz (100 micrograms/kg, i.v.). This inhibitory effect correlated positively with the time course of elevation in the estimated extracellular concentrations of SP and norepinephrine (NE) in the NRGC. Direct microinfusion of NE (50 nM/min) into the NRGC also lessened the cardiosuppressant effects of guanabenz. These circulatory and NE responses to SP were, however, significantly blunted in rats in which the noradrenergic innervation in the NRGC was depleted with N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP4) pretreatment. Microinfusion of NE into the NRGC of DSP4-pretreated animals restored the attenuation of guanabenz-induced cardiovascular suppression. These results suggest that SP may depress the activity of alpha 2-adrenoceptor at the NRGC that are involved in circulatory regulation by increasing the extracellular concentration of NE via a presynaptic modulation of noradrenergic neurotransmission.

Substance P gene expression in acute experimental colitis

We have previously shown (Gastroenterology, 101 (1991) 1211–1219), that substance P (SP) decreases early in the colon muscle layer after induction of colitis in rabbits. Since the SP content in the colonic muscle layer was unchanged by sensory denervation with capsaicin, we assume that SP is located predominantly in intrinsic neurons of the colon, and that the decrease of SP during inflammation reflects changes in intrinsic SP content. However, damage of SP neurons by inflammation would be another likely explanation for the decrease in SP content. The aim of this study was to determine SP gene expression in intrinsic (colonic muscle layer) and extrinsic (dorsal root ganglia, DRG) neurons to prove that SP transcription is preserved during colitis as an indicator of neuronal integrity. Colitis was induced in adult white New Zealand rabbits by formalin enemas (4 ml of 0.4% formalin) followed by 0.85 ml immune complex i.v. 2 h later. The animals were sacrificed 48 h after induction of colitis, and SP content and gene expression were determined by radioimmunoassay (RIA) and Northern blot analysis, respectively. Immunoreactive SP was reduced by 40% in the colon muscle layer and by 60% in the dorsal root ganglia (L 4 S 4) in the animals with colitis compared to controls without colitis. In contrast to the protein data, SP gene expression was not significantly altered in the colon muscle layer and the dorsal root ganglia 48 h after induction of inflammation compared to the control tissue. The preserved SP gene expression suggests that the intrinsic and extrinsic SP neurons are viable in this inflammatory model. The decreases of immunoreactive SP in the colonic extracts and the DRG after induction of colitis suggest that SP is released from extrinsic and intrinsic neurons during inflammation.

Time- and dose-related changes in the expression of substance P in salivary glands in response to fractionated irradiation

Purpose : The expression neuropeptides in the innervation of submandibular and parotid glands of the rats was examined 2 and 5 days after initiation of radiation treatment as well as 10 and 180 days following the termination of irradiation. Methods and Materials : The irradiation was given on 2 or 5 consecutive days with daily doses of 4–8 Gy up to a total dose of 20–40 Gy. Immunohistochemical methods were used for the demonstration of substance P (SP), calcitonin gene-related peptide (CGRP), neuropeptides Y (NPY) and, the rate-limiting enzyme in the catecholamine synthesic pathway, tyrosine hydroxylase (TH). The content of SP was also analyzed by the use of radioimmunoassay (RIA). Results : In the parenchyma of both the submandibular and the parotid glands of control animals as well as after 2 days of irradiation treatment, a few nerve fibers showing SP-like immunoreactivity (LI) were observed. A marked increase in the expression of SP in the innervation of the parenchyma in both glands was observed 10 days after cessation of radiation treatment. The number of stained nerve fibers and the intensity of fluorescence in the fibers seemed to be dose dependent because the group subjected to a total dose of 40 Gy displayed a more pronounced staining intensity than that treated with 30 Gy. These results were supported by the RIA analysis. One hundred eighty days after treatment no obvious differences in SP-expression were seen between control and irradiated animals. No acute and long-term alterations were seen with regard to the other peptides and TH. Conclusions : These results suggest that specific dose- and time-dependent changes in the expression of SP in the parenchyma of both submandibular and parotid glands occur in response to fractionated irradiation. The observations add further aspects to the tissue differences in physiological response and sensitivity to irradiation

Altered neuropeptide content and cholinergic enzymatic activity in the inflamed guinea pig jejunum during parasitism

We investigated the effects of an enteric infection with the parasitic nematode, Trichinella spiralis, on peptidergic and cholinergic neural pathways of the guinea pig jejunum. The content of the enteric neuropeptides, substance P (SP) and vasoactive intestinal peptide (VIP), and the activities of the key cholinergic enzymes, acetylcholinesterase (AChE) and choline acetyltransferase (ChAT), were measured and compared in extracts of jejunal muscularis externa (ME) obtained from uninfected jejunum and T. spiralis-inflamed jejunum. Significant decreases were detected in both SP immunoreactivity and AChE activity on days 6 and 10 postinfection (PI) in nematode-infected guinea pig jejunum compared to uninfected controls. The maximum changes observed for SP and AChE both occurred on day 10 PI and were evident as decreases of 37% and 48%, respectively, from the mean uninfected control values for SP and AChE. In contrast, VIP immunoreactivity and ChAT activity showed no significant changes during the enteric phase of T. spiralis infection. Nematode-evoked histopathological changes in jejunal tissues from infected animals were associated with significant increases in myeloperoxidase (MPO) activity, an index of inflammation intensity, which occurred on day 6 PI (885% of mean control) and day 10 PI (469% of mean control) coinciding temporally with the significant decrease in SP content and AChE activity during infection. Thus, intestinal motor disturbances observed in mammalian hosts during enteric nematode infections involve inflammation-generated changes in the neurohumoral control of smooth muscle function.

Increased levels of substance p and calcitonin gene‐related peptide in rat adjuvant arthritis: combined immunohistochemical and radioimmunoassay analysis

To analyze the occurrence of substance P (SP) and calcitonin gene-related peptide (CGRP) in ankle joints and corresponding dorsal root ganglia (L2-L6) of rats with adjuvant arthritis. Arthritis was induced by inoculation with heat-killed mycobacteria. The morphologic distribution of SP and CGRP was assessed by immunohistochemical analysis. Tissue concentrations of the neuropeptides were determined by radioimmunoassay. Neuronal CGRP-like immunoreactivity was clearly increased in the synovium and the dorsal root ganglia, whereas the increase in SP-positive structures was less pronounced. The tissue concentrations of SP and CGRP were significantly increased both in ankle joints and in dorsal root ganglia. Levels of sensory neuropeptides are increased under conditions of joint inflammation.

Two-Site Immunometric Assay For Substance P with Increased Sensitivity and Specificity

A two-site immunometric assay of the undecapeptide substance P (SP) has been developed. This assay is based on the use of two different antibodies specifically directed gist the N- and C-terminal parts of the peptide. Affinity-purified polyclonal antibodies raised against the six amino-terminal residues of the molecule were used as capture antibodies. A monoclonal antibody directed against the carboxy-terminal part of substance P, covalently coupled to the enzyme acetylcholinesterase, was used as tracer antibody. The assay is very sensitive, with a detection limit close to 3 pg/mL. Precision is also very good, with a coefficient of variation of less than 10% in the 10-250 pg/mL range. More important, the assay is fully specific for SP since cross-reactivity coefficients below 0.01% were observed with other tachykinins, SP derivatives, and SP fragments. The assay was used to measure the SP content of rat brain extracts and was validated by HPLC experiments

SP-induced contraction of airway smooth muscle in normal and allergen-sensitized rabbits: mechanism of action.

We studied the mechanisms involved in the airway smooth muscle (ASM) contraction to substance P (SP) in normal (control) and allergen-sensitized (immune) rabbits as well as immune rabbits exposed to allergen via the airways (immune challenged). Cumulative concentration-response curves to SP (1 x 10(-9) to 1 x 10(-4) M) were performed in ASM segments in the absence and presence of atropine (10(-5) M) in vitro. The maximal contractile response (g tension/g tissue) at 10(-4) M SP and ASM contractions at various concentrations of SP were expressed as means +/- SE. We found no difference in the contractile response to SP between control and immune animals. ASM segments obtained from immune-challenged rabbits were more responsive to SP. Atropine shifted to the right the concentration-response curves and decreased the maximal ASM contraction at 10(-4) M SP in all three groups; this effect, however, was greater in immune-challenged tissues. These findings demonstrate an increased contractile response to SP in immune-challenged animals mediated by a more pronounced facilitation of cholinergic neurotransmission. We conclude that the final ASM response to SP is the result of a complex interaction between direct effects on ASM and indirect effects through modulation of cholinergic neurotransmission.

Effects of progesterone on prolactin, hypothalamic beta-endorphin, hypothalamic substance P, and midbrain serotonin in guinea pigs.

Unlike rats, but similar to primates, guinea pigs exhibit prolonged function of the corpus luteum and elevated progesterone secretion after ovulation. The gonadotropins, estrogen (E) and progesterone (P) have been examined throughout the guinea pig estrous cycle. However, neither prolactin secretion nor its regulation by steroid hormones has been characterized, perhaps due to the lack of a specific radioimmunoassay. beta-Endorphin (BE), substance P (SP), and serotonin (5-HT) increase prolactin secretion in rats and monkeys. BE and SP neurons in guinea pigs and 5-HT neurons in monkeys contain progestin receptors which could mediate neuroendocrine effects of steroid hormones. Therefore, the effects of E and P on prolactin, BE, SP, and 5-HT and its metabolite 5-HIAA were examined in guinea pigs which were ovariectomized, E treated (28 days), and E+P treated (14 days E+14 days E+P). The rat NB2 lymphoma cell line was used as a bioassay for serum prolactin. BE and SP levels were measured by radioimmunoassay in four hypothalamic areas: the preoptic region (POA), the mediobasal hypothalamus (MBH), the dorsomedial hypothalamus (DMH), and the mamillary bodies (MB). 5-HT and 5-HIAA were measured in the midbrain raphe area by high-pressure liquid chromatography. E alone had little effect on serum prolactin levels, but E+P significantly increased prolactin as compared with ovariectomized controls. The BE levels increased with E treatment and remained elevated with E+P treatment in MBH and POA. The BE content was stimulated in DMH and MB by E+P treatment and not with E alone. The SP content in MBH, DMH, and MB increased in E-treated guinea pigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of substance P on the growth processes in rat thyroid lobes in vitro.

The effects of substance P (SP) on (1) 3H-thymidine incorporation into the DNA of rat thyroid lobes during a 4-hour incubation, and (2) thymidine kinase (TK, EC 2.7.1.21) activity in homogenates of rat thyroid lobes were investigated. We also examined the influence of the joint action of SP and thyrotropin (TSH) on these processes. It was shown that the effects of SP on these processes were dependent on the concentration of the neuropeptide applied. At the lowest concentration (10(-12) M), SP significantly (p < 0.0025) decreased and at the highest one (10(-6) M) it slightly increased (p < 0.09) the values of the examined growth indices. The activity of TK rose in parallel with the increase in SP concentration. SP and TSH, when used together, exerted similar decreasing effects, both on 3H-thymidine incorporation into DNA and on TK activity.

Beta-endorphin, but not substance-P, is increased by acute stress in humans.

The role of neuropeptides in the psychoneuroendocrinological stress response is largely unknown. In this study the effect of acute psychological stress on beta-endorphin and substance-P plasma concentrations was investigated and further the effect of different anxiety levels or control attributions on beta-endorphin or substance-P levels were determined. Blood samples were obtained from 47 inexperienced tandem-parachutists 2 h before, immediately after, and 1 h after a parachute jump and plasma concentrations of beta-endorphin and substance-P were analysed. Anxiety levels and control attributions were assessed by psychometric scales. Whereas substance-P concentrations seemed to be unaffected by the jump stress, there was a transient but significant increase in beta-endorphin levels immediately after jumping. However, subjects higher in state-anxiety at the point of jumping (exit) displayed higher substance-P values at all three time points compared to the "low-anxiety" jumpers. In addition, stress-induced beta-endorphin secretion was dependent on subjective control attributions.

Effects of surgical denervation on substance P and calcitonin gene-related peptide in adjuvant arthritis.

The occurrence of substance P (SP) and calcitonin gene-related peptide (CGRP) was assessed in the ankle joints and dorsal root ganglia (L2-L6) by immunohistochemistry and radioimmunoassay (RIA) after unilateral sciatic nerve section in adjuvant arthritis. Nerve section in arthritic rats had no clear-cut effect on warmth, redness, and swelling of ankle joints, but significantly affected the occurrence of SP and CGRP. Immunohistochemical analysis showed an almost complete disappearance of SP-positive fibers in the ipsilateral ankle joint in normal rats after axotomy, whereas in arthritic rats occasional SP-positive fibers remained. In dorsal root ganglia, only occasional SP-positive cells could be detected in normal and arthritic rats after axotomy. A similar but somewhat less pronounced effect of axotomy was noted for neuronal CGRP-LI. RIA showed a decrease in SP in ankle joints by 45% in normal rats and 58% in arthritic rats; the decrease in CGRP was 41% and 47%, respectively. In dorsal root ganglia, the decrease in SP after surgical denervation was 25% in normal rats and 54% in arthritic rats; the decrease in CGRP was 18% and 27%, respectively. The tissue concentrations of SP and CGRP in ipsilateral ankle joints and dorsal root ganglia were consistently correlated in normal as well as arthritic rats. The present study shows that an interruption of the nerve supply to joints cannot fully prevent the development of arthritis, although it significantly reduces the occurrence of sensory neuropeptides.

Tachykinins induce secretion of prolactin from perifused rat anterior pituitary cells by interactions with two different binding sites.

Substance P and the two other mammalian tachykinins, neurokinin A and B, are accepted to have direct regulating effects at the anterior pituitary level. We have examined the effects of substance P (SP) and neurokinin B (NKB), alone and in combination, on prolactin release from cultured anterior pituitary cells grown on collagen-coated micro beads and placed in a perfusion system. Prolactin (Prl) secretion was observed within 25 s after exposure to either secretagogue and reached a maximum within 60-80 s. Furthermore, the prolactin response induced by SP and NKB was dose-dependent. Prl secretion remained constant for up to 4 h when SP or NKB were perifused and then fell gradually towards basal levels. Simultaneous addition of submaximal concentrations of SP and NKB resulted in an additive response compared with the responses of either secretagogue alone. Continuous (8 h) perifusion with SP did not prevent a normal prolactin response by NKB or TRH. These results indicate that the tachykinins, substance P and neurokinin B, release Prl from perifused female rat anterior pituitary cells by interaction with two different receptors, possibly the NK1 and NK3 tachykinin receptor subtypes.