Immunoreactive Substance Research Articles

Ecabet sodium raises plasma levels of calcitonin gene-related peptide and substance P in healthy humans

Ecabet sodium (ecabet), a cytoprotective drug, produces an increase in mucosal blood flow. One of the gastrointestinal motility regulatory factors has been assumed to be the induction of changes in the levels of peptides (gastrin, somatostatin and motilin) in plasma. On the other hand, recently, capsaicinsensitive afferent nerves were shown to play an important role in gastric mucosal defensive mechanism. Capsaicin stimulates afferent nerves and enhances the release of calcitonin gene-related peptide (CGRP) and substance P in the stomach. We studied the effect of ecabet on human plasma gastrin-, somatostatin-, motilin-, CGRP- and substance P-like immunoreactive substance (IS) in healthy subjects. Ecabet sodium at a dose of 3.0 g, or placebo, was orally administered in five healthy males. The blood samples were taken before and at 20, 40, 60, 90, 120, 180 and 240 min after administration, subjected to extracting procedures, and submitted to a highly sensitive enzyme immunoassay system. Single administration of ecabet caused significant (P<0.05) increases in plasma CGRP-, substance P- and somatostatin-IS concentration compared with placebo. Ecabet significantly decreased plasma gastrin-IS levels compared with placebo. In this study, we hypothesized that ecabet might stimulate capsaicin-sensitive afferent nerves indirectly and improve mucosal blood flow; this might be a key mechanism underlying its gastroprotective action.

Accumulation of retinol in the liver after prolonged hyporetinolemia in the vitamin A-sufficient rat

We assessed the effects of prolonged reduction of plasma retinol concentrations (hyporetinolemia) on the distribution of tissue vitamin A (VA) and of its active compounds using a model of continuous recombinant human interleukin-6 (rhIL-6) infusion via osmotic minipumps in VA-sufficient male rats. Plasma retinol and retinol-binding protein (RBP) concentrations remained decreased and lower in rhIL-6-treated rats compared with controls from 7.5 h throughout 7 days of infusion (P < 0.001). This reduction was accompanied by a 68% increase in hepatic retinol concentration by 7 days (P < 0.05). Hepatic and renal retinyl palmitate and retinoic acid concentrations did not change, and renal megalin content remained unchanged; hepatic RBP concentrations were 41% lower in rhIL-6-treated rats compared with controls (P < 0.05). These results indicate that instead of being lost, retinol accumulated in the liver during inflammation and that hyporetinolemia was attributable to a decrease in the availability of hepatic RBP. A plausible consequence of the effect of rhIL-6-induced hyporetinolemia is that by 7 days tissues that are dependent on plasma retinol may become deprived of VA. These results have important implications in understanding the mechanism by which measles infection induces hyporetinolemia and VA deficiency of extrahepatic tissues.

Effect of gastrointestinal function regulatory Kampo medicine, Dai-kenchu-to on human plasma calcitonine gene-related peptide, substance P and somatostatin levels

Background Traditional Chinese herbal (Kampo) medicines, Dai-kenchu-to (DKCT) have been frequently used in the empirical treatment of gastrointestinal obstructions. Although it has been already reported that DKCT increases in the plasma levels of gastrointestinal peptides {vasoactive intestinal polypeptides (VIP) and motilin}, which improves feelings of abdominal coldness and gastrointestinal motility, the mechanism is not yet well understood. In this study, we examined effects of DKCT on the plasma calcitonine gene-related peptide (CGRP), substance P (SP) and somatostatin (SS) levels. Methods DKCT or placebo was orally administered to five healthy male volunteers aged 25–30 years. Venous blood samples were taken before and 20, 40, 60, 90, 120, 180 and 240 min after administration of DKCT. Peptide levels in plasma were measured using a sensitive enzyme immunoassay. Results DKCT (7.5 g) significantly increased in CGRP-like immunoreactive substances (IS) at 40 min, SP-IS at 20–60 min and SS-IS at 180–240 min compared with the each response of placebo. Conclusions In this study, DKCT raised plasma CGRP-, SP- and SS-IS levels. CGRP and SP participate in the control of mucosal blood flow in the gastrointestinal tract. Also, SS participates in the regulation of gastrointestinal motility. DKCT might regulate to the mucosal blood flow by increasing in plasma CGRP- and SP-IS levels. Besides, DKCT might improve the gastrointestinal motility by increasing in plasma SS-IS levels. Clinical Pharmacology & Therapeutics (2005) 77, P13–P13; doi: 10.1016/j.clpt.2004.11.052

Lansoprazole Raises Somatostatin, Calcitonin Gene-Related Peptide, and Substance P Levels in Healthy Human Plasma

levels were measured using a sensitive enzyme immunoassay. Compared with the response of the placebo treated group, lansoprazole caused significant (p < 0.05) increases in somatostatin-, VIP-, substance P- and CGRP-like immunoreactive substance (IS) levels at 120, 120, 90‐240, and 120 min, respectively. Furthermore, lansoprazole significantly suppressed temporary elevation (30 min) of placebo gastrin-IS levels. But lansoprazole had no effect on plasma motilin-IS levels compared with the placebo. In this study, lansoprazole raised somatostatin-IS levels and inhibited the increase in gastrin-IS levels. On the other hand, lansoprazole raised substance P- and CGRP-IS levels. Recently, capsaicin-sensitive afferent nerves have been shown to play an important role in gastric mucosal defensive mechanisms. Capsaicin stimulates afferent nerves and enhances the release of CGRP and substance P in the stomach. We hypothesize that lansoprazole might not only potently inhibit gastric acid secretion but also exert gastroprotective actions via capsaicin-sensitive afferent nerves.

Effects of Rabeprazole on Plasma Gastrointestinal Peptides in Healthy Humans

Rabeprazole sodium (rabeprazole), a proton pump inhibitor (PPI), is widely used in the treatment of peptic ulcer and gastroesophageal reflux disease. Recently, some antiulcer medicines have been elucidated pharmacologically from the viewpoint of gut-regulated hormone levels. We examined the effects of rabeprazole on plasma levels of gastrointestinal peptides [gastrin, somatostatin, motilin, substance P, and calcitonin gene-related peptide (CGRP)]. Rabeprazole at a dose of 20 mg or placebo was orally administered to 5 healthy male volunteers aged 25-30 years. Venous blood samples were taken before and 30, 60, 90, 120, 180, 240, and 360 min after administration. Plasma peptide levels were measured using a sensitive enzyme immunoassay. Compared with the response of the placebo group, rabeprazole caused significant (p < 0.05) increases in gastrin-, somatostatin-, and CGRP-like immunoreactive substance (IS) at 360, 180, and 240 min, respectively. Rabeprazole significantly decreased motilin-IS levels at 180 min compared with the placebo group. However, rabeprazole had no effect on plasma substance P-IS levels. Although rabeprazole has potent antisecretry effects, the agent may have little effect on gastrointestinal peptides.

Effects of Erythromycin on Plasma Gastrin, Somatostatin, and Motilin Levels in Healthy Volunteers and Postoperative Cancer Patients

Erythromycin, an antibiotic agent, is known to be a motilin receptor agonist. Motilin is a peptide hormone that regulates gastric motility. One of the gastrointestinal motility regulatory factors has been assumed to be the induction of changes in the levels of peptides (gastrin, somatostatin and motilin) in plasma. We have elucidated the effects of erythromycin by examining changes in the plasma levels of gastroinitestinal peptides. In this study, we investigated the effects of erythromycin on the plasma levels of gastrointestinal peptides (somatostatin, motilin, and gastrin) in healthy volunteers and patients with delayed gastric emptying (DGE). After a single oral administration, erythromycin caused a significant increase in plasma gastrin-like immunoreactive substance (IS) levels at 60 min. But the agent did not alter the levels of somatostatin- and motilin-IS. DGE is the most frequent postoperative complication after pylorus-preserving pancreatoduodenectomy. Molitin is assumed to be one important factor that influences DGE. We also examined the effects of erythromycin on the plasma motilin-IS levels of postoperative patients. The plasma motilin-IS levels were increased after 1 week of oral administration of erythromycin compared with preadministration. These results suggest that the pharmacologic effects of erythromycin in promoting gastric emptying are closely related to changes in plasma motilin-IS levels.

Comparison of the Effects of Cytoprotective Drugs on Human Plasma Adrenocorticotropic Hormone and Cortisol Levels with Continual Stress Exposure

Cetraxate hydrochloride (cetraxate), ecabet sodium (ecabet), and sulpiride, which are cytoprotective drugs, have been used to treat peptic ulcers and acute or chronic gastritis. They are reported to improve mucosal blood flow in the stomach. One of the most important factors believed to cause gastric ulcers is mental and/or physiological stress. When people feel stress, the hypothalamo-pituitary-adrenal (HPA) axis is activated. Therefore, corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and cortisol can be indicators of stress. We examined the effects of cetraxate, ecabet and sulpiride on the plasma levels of ACTH and cortisol under stress conditions by repetitive blood sampling. Venous blood samples were taken before and 20-240 min after a single administration of the drugs or a placebo. A single dose of ecabet caused significant suppression of increases in plasma ACTH-like immunoreactive substance (IS) levels at 90 to 120 min and cortisol levels at 240 min, compared with the response to placebo. Sulpiride only suppressed increases in plasma cortisol levels at 180 to 240 min, compared with the response to placebo. A single dose of cetraxate had no effect on plasma ACTH-IS and cortisol levels. Ecabet may have a modulatory effect on the HPA axis while sulpiride may have a partial modulatory effect on the HPA axis. These effects might be beneficial in stress-related disease.

Identification of Prolactin Synthesis at Early Differentiation Stage of T Cells in the Thymus

Ectopic gene expression of prolactin (PRL) has been reported in different tissues among several species. Recently, lymphocytes have also been shown to produce PRL, which is considered to be involved in the immunoregulatory system. However, when and where PRL synthesis occurs in lymphocytes remain unknown. In the present study, we attempted to examine the existence of PRL producing cells in the mouse thymus and spleen of immune tissues. We performed double fluorescence immunohistochemistry (DFIH) for mouse PRL (mPRL) and mouse CD4 that was specifically produced in T cells, and for mPRL and mouse CD25 or CD44, which specify the CD4+ precursor cells in the early stage of T cell differentiation in the thymus. Similarly, we performed DFIH for mPRL and mouse IL-4 or IFN-γ which are specifically produced in helper T cells of the mouse spleen. The immunoreactive substance for mPRL was co-localized in some CD4 or CD44 immunopositive cells in the thymus and some IL-4 or IFN-γ producing cells in the spleen. In conclusion, we found that mPRL is synthesized at the early stage of T cell differentiation.

Rapid Detection of Oleander Poisoning Using Digoxin Immunoassays

Oleander is an ornamental shrub that grows in the United States, Australia, India, Sri Lanka, China, and other parts of the world. All parts of the plant are poisonous because the presence of cardiac glycoside oleandrin. Despite its toxicity, oleander extract is used in folk medicines. Because of its structural similarity, oleandrin cross-reacts with the fluorescence polarization immunoassay (FPIA) for digoxin. We studied the potential of detecting oleandrin in serum using 5 common digoxin immunoassays (FPIA, MEIA, both from Abbott; Beckman digoxin assay on Synchron LX, Chemiluminescent assay, CLIA from Bayer Diagnostics) and a recently FDA-approved turbidimetric assay on the ADVIA 1650 analyzer (Bayer). Aliquots of drug-free and digoxin-like immunoreactive substances (DLIS)-free serum pools were supplemented with ethanol extract of oleander leaves or oleandrin (Sigma Chemicals) in amounts expected in vivo after severe overdose. We observed significant apparent digoxin concentration with FPIA, Beckman, and the new turbidimetric assay (1 mL drug-free serum supplemented with 5.0 microL of oleander extract: apparent digoxin 2.36 ng/mL by the FPIA, 0.32 ng/mL by the MEIA, 0.93 ng/mL by the Beckman, 0.82 ng/mL by the new turbidimetric assay). The CLIA showed no cross-reactivity. Similar observations were made when serum pools were supplemented with oleandrin. Because cross reactivity should be tested in the presence of the primary analyte, we supplemented serum pools prepared from patients receiving digoxin with oleander extract or oleandrin. The measured digoxin concentrations were falsely elevated with the FPIA, Beckman, and turbidimetric assays, the highest false elevation being observed with the FPIA. Surprisingly, apparent digoxin concentrations were falsely lowered when MEIA was used. Digibind neutralizes free apparent digoxin concentration in vitro in serum pools supplemented with oleander extract, and this effect can be measured by the FPIA. We conclude that FPIA is most sensitive to detect the presence of oleander in serum. In contrast, the CLIA (no cross-reactivity) should be used for monitoring digoxin in a patient receiving digoxin and self-medicated with a herbal remedy containing oleander.

Increased digitalis-like immunoreactive substances in neonatal plasma measured using fluorescence polarization immunoassay

To better define the reported increased digitalis-like immunoreactive substances (DLIS) in neonatal plasma, we studied the relation among plasma DLIS level, blank intensity (BLK-I) value at FPIA measurement and plasma total bilirubin level. The DLIS levels were measured in 10 neonates with or without jaundice and 10 infants in good health, using fluorescence polarization immunoassay (FPIA) and microparticle enzyme immunoassay (MEIA). BLK-I value and plasma total bilirubin level were also measured simultaneously. In neonates with jaundice, DLIS using FPIA, BLK-I and total bilirubin level were 0.58 +/-0.13 ng/mL, 2598 +/- 408, and 17.98 +/- 1.13 mg/dL, respectively, before phototherapy, and 0.33 +/-0.06 ng/mL, 1886 +/- 237, and 15.16 +/- 2.07 mg/dL after phototherapy. Corresponding values in neonates without jaundice were (DLIS: 0.34 +/-0.04 ng/mL; BLK-I: 1,764 +/- 278; total bilirubin: 10.37 +/- 4.54 mg/dL); in healthy infants (0.12 +/-0.06 ng/mL, 400.7 +/- 4.6 and 0.42 +/- 0.13 mg/dL, respectively) and in healthy volunteers (0.10 +/-0.07 ng/mL, 403.1 +/- 8.4, and 0.58 +/- 0.30 mg/dL, respectively). Using MEIA, DLIS was not detected in 10 neonates, 10 infants and 20 healthy volunteers. A fluorescent compound related to bilirubin increased the BLK-I value in the measurement of neonatal plasma using FPIA. The fluorescence was not the result of endogenous digitalis-like factors.

Immunolocalization of VR1 and VRL1 in rat larynx

Immunoreactivity for vanilloid receptor subtype 1 (VR1) and its analogue vanilloid receptor-like protein 1 (VRL1) were examined in combination with immunoreactivity for substance P (SP) and calcitonin gene-related peptide (CGRP) in the rat larynx. VR1 and VRL1 immunoreactivity were observed in the intraepithelial free nerve endings, subepithelial nerve plexus and laryngeal epithelial cells. Most of VR1 immunoreactive nerves were also immunoreactive for SP or CGRP. VR1 immunoreactive intraepithelial nerve endings may be laryngeal nociceptors.

Selective loss of calcitonin gene-related Peptide-expressing primary sensory neurons of the a-cell phenotype in early experimental diabetes.

To evaluate the possible role of neuropeptide immunoreactive primary sensory neurons on the development of nociceptive dysfunction in diabetes, the absolute numbers of immunoreactive substance P and calcitonin gene-related peptide (CGRP) dorsal root ganglion (DRG) cell bodies were estimated in diabetic and nondiabetic BALB/C (p75(+/+)) and p75 receptor knockout (p75(-/-)) mice with unilateral sciatic nerve crush. The total numbers of immunoreactive substance P A-cells, substance P B-cells, CGRP A-cells, and CGRP B-cells in L5DRG were estimated using semithick consecutive sections and the optical fractionator. After 4 weeks of streptozotocin-induced diabetes, the number of immunoreactive CGRP A-cells was reduced from 692 +/- 122 to 489 +/- 125 (P = 0.004) in p75(+/+) mice on the noncrushed side. In p75(-/-) mice, there was no such effect of diabetes on the immunoreactive CGRP A-cell number. In p75(+/+) and p75(-/-) mice, there was no effect of diabetes on the immunoreactive CGRP B-cell number, nor was there any effect of diabetes on the immunoreactive substance P B-cell number. Sciatic nerve crush was associated with a substantial loss of L5DRG B-cells in diabetic and nondiabetic p75(+/+) mice and with substantial loss of immunoreactive substance P cells in diabetic p75(+/+) mice. In diabetic and nondiabetic p75(-/-) mice, there was no crush effect on neuropeptide expression. It is concluded that experimental diabetes in the mouse is associated with loss of immunoreactive CGRP primary sensory neurons of the A-cell phenotype, that this loss could play a role for the touch-evoked nociception in the model, and that the neuronal immunoreactive CGRP abnormality possibly is mediated by activation of the p75 neurotrophin receptor.

Isoflavonoids in the rutaceae family: 1. Fortunella obovata, Murraya paniculata and four Citrus species

Several types of compounds with immunoreactivity similar to isoflavonoids were detected in water: ethanol extracts of leaves of Fortunella obovata Hort. ex Tanaka, Murraya paniculata Jack. and four Citrus species, namely C. aurantium L, C. grandis Osbeck, C. limonia Osbeck., and C. sinensis Osbeck (Rutaceae). The chromatographic mobilities of the immunoreactive substances were compared with those of authentic standards, revealing a spectrum of isoflavonoid metabolites in all plants studied. Aglycones as well as glycosides were recognized, namely daidzin, genistin, daidzein, genistein, formononetin, biochanin A, prunetin, and several incompletely characterized isoflavonoids. A subsequent HPLC-MS study verified the identities of the main immunoreactive isoflavonoids and established the identities of several others, viz. glycitein, glycitin, ononin and sissotrin, including the malonylated and acetylated isoflavonoid glucosides. The estimated content of the individual immunoreactive entities ranged from a few microg to about 2 mg/kg (dry weight). It is concluded that the isoflavonoid metabolic pathway is present throughout the Rutaceae family.

Nerve growth factor treatment enhances release of immunoreactive calcitonin gene-related peptide but not substance P from spinal dorsal horn slices in rats

The effect of systemic nerve growth factor (NGF) on neuropeptide content and capsaicin-evoked release of neuropeptide from in vitro spinal cord dorsal horn slices was examined. Rats were injected subcutaneously every other day with murine NGF (mNGF) 1 mg/kg or saline for 7 days, or mNGF 0.1/kg, mNGF 1 mg/kg or saline for 13 days. Lumbar dorsal horn slices of the rat spinal cord from all groups showed a significant increase in immunoreactive calcitonin gene-related peptide (CGRP) release upon exposure to capsaicin. This release was enhanced in rats pretreated with mNGF 1 mg/kg for 7 days, but not after 13 days. No enhancement was seen after 7 or 13 days in any treatment group for immunoreactive substance P release. Upon examination of neuropeptide content in dorsal horn, no significant differences were noted between treatment groups. The increased iCGRP release from dorsal horn slices suggests a preferential release of CGRP and provides further evidence that NGF indirectly plays a role in the modulation of inflammation through the regulation of neuropeptide release.

Substance P release in the cat spinal cord upon afferent C-fibre stimulation is not attenuated by clonidine at analgesic doses*1

In anaesthetized cats, antibody microprobes were used to measure the release of immunoreactive substance P (irSP) in the lumbar dorsal horn during electrical stimulation of primary afferent fibres at intensities suprathreshold for unmyelinated fibres. Release of irSP was detected in the region of the superficial dorsal horn. This evoked release was not reduced by clonidine hydrochloride, administered intravenously or by superfusion of the dorsal cord surface. Microprobes inserted during cord superfusion with lignocaine hydrochloride detected less irSP along their entire length, including in the region of evoked release. The results suggest that the analgesic action of clonidine does not involve reduced release of SP from the central terminals of nociceptors in the spinal cord.

Substance P release in the cat spinal cord upon afferent C-fibre stimulation is not attenuated by clonidine at analgesic doses

In anaesthetized cats, antibody microprobes were used to measure the release of immunoreactive substance P (irSP) in the lumbar dorsal horn during electrical stimulation of primary afferent fibres at intensities suprathreshold for unmyelinated fibres. Release of irSP was detected in the region of the superficial dorsal horn. This evoked release was not reduced by clonidine hydrochloride, administered intravenously or by superfusion of the dorsal cord surface. Microprobes inserted during cord superfusion with lignocaine hydrochloride detected less irSP along their entire length, including in the region of evoked release. The results suggest that the analgesic action of clonidine does not involve reduced release of SP from the central terminals of nociceptors in the spinal cord.

Effects of Ninjin-to on Levels of Calcitonin Gene-Related Peptide and Substance P in Human Plasma

The herbal medicine Ninjin-to has been used for the treatment of gastroenteritis, esogastritis, gastric atony, gastrectasis, vomiting, and anorexia. One of the mechanisms of the empirical effects is assumed to be due to local changes in neuropeptide levels. Sensory afferent neurons in the gastrointestinal mucosa regulate neuropeptides [calcitonin gene-related peptide (CGRP), substance P, etc.], which play various physiologic roles. To determine whether the pharmacologic effects of Ninjin-to on the gastrointestine are due to changes in gastrointestinal mucosa regulatory peptide levels, we examined the effects of Ninjin-to on the levels of CGRP-like immunoreactive substances (IS) and substance P-IS in plasma taken from five healthy subjects. A single oral administration of 6.0 g of Ninjin-to caused significant increases in plasma CGRP-IS at 40 min and 60 min, and in substance P-IS levels at 90 min, compared with a placebo group. These results may indicate that the pharmacologic actions of Ninjin-to are closely related to changes in CGRP-IS and substance P-IS levels.

Dai-kenchu-to Raises Levels of Calcitonin Gene-Related Peptide and Substance P in Human Plasma

Sensory afferent neurons in the gastrointestinal mucosa regulate neuropeptides [calcitonin gene-related peptide (CGRP), substance P, etc.], which play various physiologic roles and are gastroprotective. To determine whether the pharmacologic effects of Dai-kenchu-to (DKCT) on the gastrointestine are due to changes in gastrointestinal mucosa regulatory peptide levels, we examined the effects of the DKCT on the levels of CGRP-like immunoreactive substances (IS) and substance P-IS in plasma taken from five healthy subjects. A single oral administration of DKCT 7.5 g caused significant increases in plasma CGRP-IS at 40 min, and in substance P-IS levels at 20 and 60 min, compared with a placebo group. The present study may indicate that the pharmacologic action of DKCT is closely related to changes in CGRP- and substance P-IS levels.

Myocardial ischemia induces the release of substance P from cardiac afferent neurons in rat thoracic spinal cord.

Antibody-coated microprobes were inserted into the thoracic (T3-4) spinal cord in urethane-anesthetized Sprague-Dawley rats to detect the differences in the release of immunoreactive substance P-like (irSP) substances in response to differential activation of cardiac nociceptive sensory neurons (CNAN). CNAN were stimulated either by intrapericardial infusion of an inflammatory ischemic exudate solution (IES) containing algogenic substances (i.e., 10 mM each of adenosine, bradykinin, prostaglandin E2, and 5-hydroxytryptamine), or by transient occlusion of the left anterior descending coronary artery (CoAO). There was widespread basal release of irSP from the thoracic spinal cord. Stimulation of the CNAN by IES did not alter the pattern of release of irSP. Conversely, CoAO augmented the release of irSP from T3-4 spinal segments from laminae I-VII. This CoAO-induced irSP release was eliminated after thoracic dorsal rhizotomy. These results indicate that heterogeneous activation of cardiac afferents, as with focal coronary artery occlusion, represents an optimum input for activation of the cardiac neuronal hierarchy and for the resultant perception of angina. Excessive stimulation of cardiac nociceptive afferent neurons elicited during regional coronary artery occlusion involves the release of SP in the thoracic spinal cord and suggests that local spinal cord release of SP may be involved in the neural signaling of angina.

Pregnancy-induced changes in substance P and neurokinin 1 receptor (NK1-R) expression in the rat uterus

Adrenergic nerve fibres of the mammalian uterus degenerate during pregnancy. The behaviour of peptidergic fibres, such as substance P-positive fibres and of its preferred neurokinin 1 receptor (NK1-R), is poorly studied in the pregnant rat uterus. The present study analysed the changes in substance P immunoreactivity and in the expression of NK1-R protein in the uterus of non-pregnant, pregnant (days 7, 14 and 21) and postpartum rats (days 1, 8 and 22) by immunohistology, dot blot analysis and western blot analysis. In non-pregnant rats, substance P-positive fibres were localized to the myometrium; these fibres progressively disappeared during gestation and were almost absent at term (day 21). At day 22 post partum, substance P-positive fibres had recovered to numbers comparable with those in the non-pregnant uterus. Dot blot analysis revealed a significant decrease in the immunoreactivity of substance P in the uterus at mid-pregnancy (day 14) and especially at term. Expression of the NK1-R protein showed a progressive increase throughout pregnancy reaching a peak on day 1 post partum; downregulation of NK1-R protein occurred on day 8 post partum. The low and high expressions of NK1-R protein were coincident with a large number of eosinophils and almost no eosinophils in the uterus at oestrus and at term, respectively. It was concluded that substance P immunoreactivity is inversely correlated with NK1-R protein expression in the pregnant and postpartum uterus. The marked upregulation of NK1-R protein at term and after birth indicates that the NK1-R may be involved in the complex regulation of labour and postpartum physiology. However, it is likely that the NK1-protein is not involved in the recruitment of eosinophils into the uterus at oestrus.