Evidence for coexistence of choline acetyltransferase (ChAT)- and calcitonin gene-related peptide (CGRP)-immunoreactivity in the thoracolumbar and sacral spinal cord neurons of the pig.

Previous immunocytochemical studies provided conflicting data concerning occurrence of the CGRP-immunoreactive neuronal cell bodies in the porcine spinal cord. In the present study, we have investigated expression of the CGRP and its possible coexpression with ChAT in the gray matter of the thoracic, lumbar and sacral spinal cord of the pig. Our study revealed a large number of CGRP-immunoreactive cells in the motor nucleus of the ventral horn, and less and singe perikarya intermediolateral and intermediomedial nuclei, respectively. Double staining immunocytochemistry, depending on the cross-section level, disclosed the highest ChAT/CGRP colocalization subsequently in the motor nucleus of the ventral horn, then in the intermediolateral and intermediomedial nucleus. Our data provide morphological evidence confirming expression of CGRP in the porcine spinal nuclei while its coexpression in cholinergic neurons suggests that CGRP may play a role in modulation of the spinal cholinergic transmission.

Distribution and chemical coding of intramural neurons in the porcine ileum during proliferative enteropathy.

Enteric neurons are highly adaptive in their response to various pathological processes including inflammation, so the aim of this study was to describe the chemical coding of neurons in the ileal intramural ganglia in porcine proliferative enteropathy (PPE). Accordingly, juvenile Large White Polish pigs with clinically diagnosed Lawsonia intracellularis infection (PPE; n=3) and a group of uninfected controls (C; n=3) were studied. Ileal tissue from each animal was processed for dual-labelling immunofluorescence using antiserum specific for protein gene product 9.5 (PGP 9.5) in combination with antiserum to one of: vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), somatostatin (SOM), neuropeptide Y (NPY) or galanin (GAL). In infected pigs, enteric neurons were found in ganglia located within three intramural plexuses: inner submucosal (ISP), outer submucosal (OSP) and myenteric (MP). Immunofluorescence labelling revealed increases in the number of neurons containing GAL, SOM, VIP and CGRP in pigs with PPE. Neuropeptides may therefore have an important role in the function of porcine enteric local nerve circuits under pathological conditions, when the nervous system is stressed, challenged or afflicted by disease such as PPE. However, further studies are required to determine the exact physiological relevance of the observed adaptive changes.

Immunologic status in pediatric cardiosurgical patients with chylothorax.

OBJECTIVE: The impact of lymphocyte and immunoglobulin loss on immunologic status has not been extensively studied in children with chylothorax. The purpose of this study was to evaluate immunologic profile of pediatric cardiosurgical patients who developed infection while suffering from prolonged postoperative chylothorax. METHODS: We retrospectively reviewed immunologic findings in 16 pediatric cardiac patients with post-operative chylothorax persisting ?7 days. Patients were on total parenteral nutrition, received colloides for replacement of chylous losses, and antibiotics and/or antimycotics for treatment of infection. Immunologic evaluation included immunoglobulin levels, cellular immunity, and phagocytic activity. For every parameter z-score was calculated according to age-dependent nomograms and t-test was used to compare z-score distribution with normal distribution. RESULTS: The immunoglobulin (IgG, IgM, and IgA) levels did not significantly differ from normal values, although 25% patients had IgG levels below normal range. The relative and absolute counts of peripheral blood lymphocytes were lower (p < 0.001) than normal values. Absolute numbers of blood B-lymphocytes (CD19+), T-lymphocytes (CD3+), helper/inducer T-cells (CD4+), and suppressor/cytotoxic T-cells (CD8+) were also below normal range (p < 0.001); however, their relative percentages and a CD4+/CD8+ ratio were within normal limits. The percentage and absolute number of natural killer cells (CD16+), phagocytic and metabolic activity of polymorphonuclear leukocytes did not differ from normal values. CONCLUSIONS: Persisting chylothorax results in B-cell and T-cell lymphopenia with proportional decline of CD4+ and CD8+ cells. Hypogammaglobulinemia observed in other studies has not been detected in this series probably due to administered plasma. Effects of these immunologic alterations on development of infection are unknown (Tab. 2, Ref. 13).

Arrhythmias in congenital heart defects.

AIM OF THE STUDY: Evaluation of the incidence and severity of late arrhythmias in patients with predisposing congenital heart defects--either due to the anatomy of the defect itself or as a result of a particular type of surgical intervention. PATIENTS AND METHODS: In a retrospective long-term study authors analyzed 158 patients (divided into 5 groups) with congenital heart defects after surgical correction. Evaluated were: the incidence of rhythm disturbances, the type of arrhythmia and the need for medication or intervention. RESULTS: The most rhythm disturbances occurred in patients after physiological correction of D-transposition of the great arteries (68.5%) and these patients also mostly needed medication or pacemaker implantation; followed were by patients with hypoplastic left heart syndrome after Fontan procedure (40%), then were patients after long-term correction of tetralogy of Fallot (31.1%), atrial septal defect sinus venosus type with partial anomalous pulmonary venous return after Warden correction (25.7%) and congenitally corrected L-transposition of the great arteries (25 %). Most of these arrhythmias were asymptomatic and there was no need to treat them. There was an increased incidence of arrhythmias with time (p < 0.05). DISCUSSION: During childhood in patients after surgical correction late arrhythmias mostly do not represent a severe problem, but with time, when reaching adulthood, this may be an issue. It is therefore very important to understand the anatomy, physiology and the arrhythmogenic substrate of every high risk congenital heart defect (Tab. 2, Fig. 6, Ref. 10).

Re-examination of the topographical localization of facial nucleus in the pig.

Previous publications have provided different descriptions of the topographical organization of the facial nucleus of the pig. Since swine is used in biomedical research due to its embryological, anatomical and physiological similarities to human, we have reinvestigated the anatomical organization of the facial nucleus with application of fluorescent retrograde tracer Fast Blue, antibody to choline acetyltransferase and acetylcholinesterase histochemistry. Our findings demonstrate that in the porcine medulla facial motoneurons constitute a large cellular group occupying the ventro-lateral medulla. The neuronal group is interposed rostro-caudally between the superior and inferior olive, and located ventro-medially to the spinal nucleus of the trigeminal nerve. The present results clarify the anatomical description of this important brain stem nucleus in the pig.

Immunohistochemical characterization of neurons in the porcine ciliary ganglion.

The present study was aimed at disclosing the chemical coding of nerve structures in the porcine ciliary ganglion (CG) using immunohistochemical methods. The substances under investigation included markers of "classical" neurotransmitters, choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DbetaH) as well as neuropeptides, somatostatin (SOM), galanin (GAL), substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY). Immunoreactivity to ChAT and VAChT was found virtually in all the neuronal somata and in numerous intraganglionic, varicose nerve fibres which often formed basket-like formations around the nerve cell bodies. Many CG neurons contained immunoreactivity for SOM (46%) or GAL (29%). Interestingly, a small number (approx. 1%) of the cholinergic somata stained for TH but not for DbetaH; nevertheless, some extra- and intraganglionic nerve fibres displayed immunoreactivity for DbetaH or TH. The CG perikarya stained neither for vasoactive intestinal polypeptide (VIP) nor for neuropeptide Y (NPY), but some NPY- or VIP-positive nerve terminals were observed within nerve bundles distributed outside the ganglion. SP- and CGRP-immunoreactivity was found in some intraganglionic nerve fibres only. The present study revealed that the porcine CG consists of cholinergic neurons many of which contain SOM and GAL. Thus, it can be assumed that in the pig, these neuropeptides are involved, complementary to acetylocholine, in the parasympathetic postganglionic nerve pathway to structures of the eye including the ciliary and iris sphincter muscles.

The influence of experimental ileitis on the neuropeptide coding of enteric neurons in the pig.

In the present study, both the ELISA test and immunohistochemical staining were used to investigate the influence of artificially induced ileitis on the chemical coding of enteric neurons in the pig. The ileum wall in experimental (E) pigs was injected in multiple sites with 4% paraformaldehyde to induce inflammation, while in the control (C) animals, the organ was injected with 0.1M phosphate buffer (pH 7.4). Three days after ileitis induction, samples of ileum wall from all the animals were evaluated for VIP, SP, CGRP, NPY, GAL and SOM concentration (ELISA test) and the expression of these biologically active substances by the enteric neurons (immunohistochemical staining). Quantitative results showed that ileitis decreased tissue concentration of VIP, CGRP and SOM but increased tissue concentration of SP, NPY and GAL. Immunochemistry revealed that in both the experimental and control pigs, VIP-positive (VIP+) nerve fibers supplied mainly ileal blood vessels, and the labeled pericarya were located in the inner (ISP) and outer submucous plexus (OSP). SP+ and CGRP+ nerve terminals were found in both the mucous and muscular membrane, while the labeled pericarya were found in ISP, OSP and myenteric plexus (MP). In both C and E pigs, the very few nerve terminals containing NPY and SOM were located mainly in the mucous membrane. NPY- or/and SOM-immunopositive nerve cell bodies were found in ISP, OSP and MP. GAL+ nerve fibers supplied all layers of the ileum and were most numerous in the muscular membrane, while the labeled pericarya were present in all the enteric plexuses. The present results suggest that enteric neurons are highly plastic in their response to inflammation.

Met5-enkephalin- and Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactive nerve fibers in the pig female reproductive system.

The localization of Met5-enkephalin (ME) immunoreactivity in the female genital organs of the rat, guinea pig and pig was studied by indirect immunofluorescence method. In the rat and guinea pig, no ME immunoreactivity was observed in the uterus, fallopian tube or ovary. In the pig uterus and fallopian tube ME-immunoreactive nerve fibers were observed in muscular and submucose layers as well as around the blood vessels. In the pig ovary, ME immunoreactivity was localized in nerve fibers in medullary and cortical parts of the organ. Met5-enkephalin-Arg6-Gly7-Leu8 (MEAGL) immunoreactivity was also studied in the pig uterus, where its distribution was similar to that of ME. The present results suggest that the pig genital organs receive innervation by nerve fibers containing proenkephalin A-derived peptides, which may have a role in modulation of neurotransmission in these organs.

Immunocytochemical demonstration of neuropeptide Y and luteinizing hormone-releasing hormone-immunoreactive structures in the organum vasculosum laminae terminalis of juvenile gilts.

Immunocytochemical investigations on the immature gilt organum vasculosum laminae terminalis showed extensive neuropeptide Y- and luteinizing hormone-releasing hormone-immunoreactive innervation of the organ. The luteinizing hormone-releasing hormone-containing varicose fibers ran along the organum vasculosum laminae terminalis in close association with blood vessels. The nerve processes originating from well-stained luteinizing hormone-releasing hormone-immunoreactive perikarya were distributed around the organum vasculosum laminae terminalis. The matrix of the organum vasculosum laminae terminalis was abundantly supplied by neuropeptide Y-immunoreactive varicose fibers. Numerous neuropeptide Y-immunoreactive terminals seemed to penetrate the ependymal lining of the organ. From these observations, it is concluded that there are favorable morphological conditions for secretion of neuropeptide Y into the cerebrospinal fluid of the third ventricle and release of luteinizing hormone-releasing hormone into fenestrated capillaries of the organ.

Distribution of somatostatin- and neuropeptide Y-immunoreactive nerve fibers in the porcine female reproductive system.

The localization and distribution of somatostatin and neuropeptide Y were studied in the porcine female reproductive system with the indirect immunofluorescence technique. Somatostatin-immunoreactive nerve fibers were observed in different parts of the ovary and in the muscular membrane of the uterus as well as in the mesosalphinx. Somatostatin-immunoreactive neurons were detected in the inferior mesenteric ganglion. Neuropeptide Y immunoreactivity was present in a large number of nerve fibers distributed in different regions of the uterus, oviduct and ovary. The present results suggest that the porcine female genital organs receive innervation by somatostatin- and neuropeptide Y-containing nerve fibers, but their exact functional role remains to be established.

5-Hydroxytryptamine-immunoreactive nerve fibers in the rat and porcine prevertebral sympathetic ganglia: effect of precursor loading and relation to catecholaminergic neurons.

Localization of 5-hydroxytryptamine immunoreactivity was studied in the rat coeliac-superior mesenteric ganglion complex and in the porcine superior and inferior mesenteric ganglia by the indirect immunofluorescence technique. In normal rats, only 5-hydroxytryptamine immunoreactive SIF cells were seen in the coeliac-superior mesenteric ganglion complex. In the rats, pretreated with a 5-hydroxytryptamine precursor, L-tryptophan, and with a monoamine oxidase inhibitor, nialamide, a large number of 5-hydroxytryptamine-immunoreactive nerve fiber terminals were detected. In normal porcine superior and inferior mesenteric ganglia, intense 5-hydroxytryptamine immunoreactivity was found in numerous nerve fibers which were located around tyrosine hydroxylase-immunoreactive principal neurons. The origin and function of these fibers are discussed.

Immunohistochemical colocalization of Met5-enkephalin-Arg6-Gly7-Leu8 with tyrosine hydroxylase in neurons of the porcine inferior mesenteric and coeliac-superior mesenteric ganglia.

The localization and distribution of the proenkephalin A-derived octapeptide Met5-enkephalin-Arg6-Gly7-Leu8 (MEAGL) was studied in the porcine sympathetic prevertebral ganglia by using the indirect immunofluorescence technique. In the inferior mesenteric ganglion, a small population of principal neurons and SIF cells contained intense MEAGL immunoreactivity. A very dense plexus of MEAGL-immunoreactive nerve fibers was distributed in different parts of the ganglion. A low number of MEAGL- immunoreactive principal ganglion cells and SIF cells, as well as numerous nerve fibers were also present in the coeliac-superior mesenteric ganglion. Double staining experiments indicated that the MEAGL-immunoreactive principal neurons in the inferior mesenteric and coeliac-superior mesenteric ganglia also contained tyrosine hydroxylase immunoreactivity.

Noradrenergic and peptidergic innervation of the testis and epididymis in the male pig.

This study was designed to investigate the distribution of noradrenaline (NE)- and peptide-containing nerves in the testis of the boar. Testes, as well as caput and cauda epididymides from five 5 week-old and 3 adult boars were sectioned and immunostained with antisera to tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), Leu-enkephalin (L-ENK) and somatostatin (SOM). In addition, double-labelling immunofluorescence method was used to disclose the pattern of co-existence of these substances in the nerve fibres. The most abundant innervation was found in the cauda epididymidis, and the density of the nerves distinctly decreased towards the caput epididymidis. The testicular parenchyma and caput epididymidis was supplied with very scattered TH-, D beta H-, NPY-, VIP-, SP-, CGRP-, L-ENK- and SOM-containing nerve fibres. The present study has demonstrated for the first time the existence of CGRP, L-ENK and SOM in nerve fibres innervating the testis of a vertebrate species. Different subpopulations of nerve fibres, including TH+/D beta H+, D beta H+/NPY+, D beta H-/NPY+, D beta H+/NPY-, D beta H+/VIP+, D beta H-/VIP+, D beta H+/SP+, D beta H-/SP+, D beta H+/L-ENK+ D beta H-/L-ENK+, D beta H+/SOM+ and D beta H-/SOM+, were localized and documented.

Adrenergic innervation of the epididymis, vas deferens, accessory genital glands and urethra in the boar.

The distribution of adrenergic nerve fibres in the urethra and some organs of the male reproductive system of juvenile boars was studied by applying the glioxylic acid-induced fluorescence method. Adrenergic nerve fibres were found in all structures examined, with particular relation to their muscular coats and arterial blood vessels. Ductuli efferentes and ductus epididymidis from the caput region were weakly innervated. The number of adrenergic nerve fibres innervating the ductus epididymidis gradually increased towards the cauda epididymidis, being greatest in this region. The vas deferens was very well supplied with these nerves. The seminal vesicle was found to be the richest innervated organ in comparison with other accessory genital glands. The body of the prostate was slightly weaker innervated, while the disseminated part of the prostate as well as the bulbourethral gland comprised the smallest number of adrenergic nerve fibres. The urethra received a very dense adrenergic nerve supply. The pelvic part of the urogenital duct possessed a moderate number of adrenergic nerves, which supplied not only the muscular membrane but also venous sinuses of the mucosa. In a close vicinity to the internal genital organs numerous ganglia containing both adrenergic and non-adrenergic nerve cell bodies were observed. The presented data suggest that adrenergic innervation may be deeply involved in the control of blood flow and motor function of the organs studied, and that a part of adrenergic nerve fibers innervating these structures may originate from pelvic ganglia.

Immunohistochemical localization of neuropeptides in nerve fibers of the porcine vagina and uterine cervix.

The distribution of vasoactive intestinal polypeptide, substance P, Met5-enkephalin-Arg6-Gly7-Leu8, neuropeptide Y, calcitonin gene-related peptide and bombesin/gastrin releasing peptide was studied immunohistochemically in nerve fibres supplying the vagina and uterine cervix of sexually immature pigs. Nerves containing neuropeptide Y- and Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactivity were particularly numerous in the uterine cervix and vagina. Vasoactive intestinal polypeptide- and and calcitonin gene-related peptide-containing nerves were less numerous, whereas the bombesin/gastrin releasing peptide- and substance P-immunoreactive nerves were scarce in these organs. Generally, the immunoreactive fibres, nerve networks or bundles were distributed under the serous membrane, among smooth muscle of muscular membranes, in the submucosal layer and under the luminar epithelium of the uterine cervix and vagina.

Neuropeptides in the porcine coeliac-superior mesenteric ganglion.

The occurrence and distribution of tyrosine hydroxylase, neuropeptide Y, somatostatin, Met5-enkephalin-Arg6-Gly7-Leu8, vasoactive intestinal polypeptide, substance P, calcitonin gene-related peptide, bombesin gastrin releasing peptide and galanin were immunohistochemically studied in the perikarya and nerve fibres of the porcine coeliac-superior mesenteric ganglion of untreated juvenile pigs. Subpopulations of neurons containing immunoreactivities to tyrosine hydroxylase, neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8, somatostatin, vasoactive intestinal polypeptide and galanin were disclosed in the studied ganglion, whereas principal ganglionic cells were non-immunoreactive for other investigated peptides. Double-immunofluorescence and analysis of consecutive sections revealed a partial colocalization of tyrosine hydroxylase and neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8 and somatostatin, whereas immunoreactivity to vasoactive intestinal polypeptide and/or to neuropeptide Y was found in non-noradrenergic neurons in this ganglion. All of neuropeptides studied were found in nerve fibres in this ganglion. The results of this study were compared with those of previous studies performed on other species.

Immunohistochemical study of the existence and coexistence of catecholamine synthesizing enzymes and some neuropeptides in perivascular nerve fibres of the main thoraco-cranial arteries in the pig.

The presence and pattern of coexistence of catecholamine-synthesizing enzymes and some neuropeptides in nerve fibres supplying thoraco-cranial arteries of the sexually immature gilts were investigated in whole mount preparations. The studied substances included: tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H) (as markers of catecholaminergic nerve fibres), neuropeptide Y (NPY), Leu5-enkephalin (LENK), vasoactive intestinal polipeptide (VIP), calcitonin gene-related peptide (CGRP), substance P (SP), galanin (GAL), somatostatin (SOM) and serotonin (5-HT). The arteries were found to be richly supplied by TH/D beta H-immunoreactive (TH/D beta H) nerve fibers. Of the neuropeptides studied, NPY (rich innervation), LENK (moderate innervation), VIP (moderate innervation) and CGRP, SP, GAL (only a few nerve fibres) were detected in periarterial nerves. The following patterns of coexistence of the studied substances were found: TH+/D beta H+, TH+/D beta H+/NPY+, TH+/D beta H+/LENK+, TH-/D beta H-/NPY+, TH-/D beta H-/VIP+, TH+/D beta H+/VIP+ (only a few nerve fibres in the cerebral arteries), LENK+/NPY+, LENK-/NPY+, LENK+/NPY-. No SOM and 5-HT-positive structures were observed in the porcine blood vessels.

Occurrence and coexistence of some neuropeptides in nerve fibers supplying the bovine ovary and its extrinsic blood vessels.

Single- and double-labelling immunofluorescence was applied to investigate the occurrence and coexistence of vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and substance P (SP) in nerve fibers supplying the bovine ovary as well as its extrinsic blood vessels. We also studied the coexistence of these neuropeptides with tyrosine hydroxylase (TH) and neuropeptide Y (NPY) to determine features of "chemical coding" of nerve fibers innervating this organ. The bovine ovary was found to be supplied by VIP- and SP-immunoreactive (VIP- and SP-IR) nerve fibers. No intraovarian CGRP-IR nerve terminals were detected. Immunoreactivity to VIP was shown in a moderate number of nerve fibers occurring in all ovarian regions - ovarian hilus, cortex and medulla. VIP-IR nerves were found to innervate blood vessels and ovarian follicles, especially primordial and primary ones. Immunoreactivity to SP was detected only in solitary intraovarian nerve fibers which were also encountered in all ovarian regions, but with special preference to ovarian medulla, where they innervated blood vessels. SP-IR nerve fibres were sometimes observed in close vicinity to primordial and primary follicles. Double-labelling immunofluorescence revealed that some VIP-IR nerve fibres supplying intraovarian blood vessels were simultaneously TH- or NPY-IR. Small numbers of VIP-IR fibres contained immunoreactivity to SP. All SP-IR nerve terminals were TH-or NPY-negative. The ovarian artery as well as arteries of the paraovarian plexus were moderately supplied with VIP-, or by single CGRP- or SP-IR nerve fibres. Veins were innervated by only solitary nerve terminals containing these neuropeptides. The following patterns of coexistence were determined in nerve fibres supplying the investigated blood vessels: VIP, VIP/CGRP, VIP/SP, CGRP/SP, VIP/NPY. No distinct differences in the innervation of the ovary and its extrinsic blood vessels between juvenile and adult cows were found.

Distribution of some neuropeptides in the porcine inferior mesenteric ganglion.

The occurrence and distribution of vasoactive intestinal polypeptide, substance P, and bombesin/gastrin-releasing peptide in neuronal cell bodies and nerve fibers of the porcine inferior mesenteric ganglion were studied with the indirect immunohistochemical technique. Of all substances studied only vasoactive intestinal polypeptide was found in principal ganglionic neurons. The presence of nerve fibers immunoreactive to vasoactive intestinal polypeptide, bombesin/gastrin-releasing peptide, and substance P was also found in the ganglion. There were differences in the pattern of distribution and density of the nerve fibers immunoreactive to the particular peptides. Fibers containing vasoactive intestinal polypeptide and bombesin/gastrin-releasing peptide were numerous, while fibers containing substance P were comparatively scarce. The present results revealed both similarities and specific differences in the occurrence and localisation of various neuropeptides in the porcine inferior mesenteric ganglion in comparison with that of other mammalian species.

Peptidergic innervation of the porcine oviduct studied by double-labelling immunohistochemistry.

Neurochemical coding of nerve fibres supplying the porcine oviduct was studied by means of double-labelling immunofluorescence. Immunoreactivities to rate-limiting enzymes of catecholamine synthesis, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DbetaH), were found in numerous oviductal nerve fibres. All TH-immunoreactive (TH-IR) nerve processes were simultaneously DbetaH-IR. This subset of nerves was considered to be sympathetic noradrenergic. In addition to noradrenaline, many axons additionally exhibited immunoreactivity to neuropeptide Y (NPY), or leu5-enkephalin (Leu-ENK). Small numbers of somatostatin- (SOM-) or vasoactive intestinal polypeptide-immunoreactive (VIP-IR) fibres, sometimes coexpressing TH/DbetaH-immunoreactivity, supplied the porcine oviduct. Substance P- (SP- ) and/or calcitonin gene-related peptide-immunoreactive (CGRP-IR) nerve fibres were only sporadically found. Although these nerves did not contain TH/DbetaH-immunoreactivity, they often ran in close vicinity to TH/DbetaH-IR axons, forming together thin nerve bundles. All the above mentioned subpopulations of nerve fibres were found throughout the entire length of the oviduct being mainly related to the vascular and non-vascular smooth myocytes. However, some of the putative afferent (i.e. SP- or CGRP-IR) or parasympathetic efferent (i.e. VIP- or NPY-IR but TH/DbetaH-immunonegative) axons were located beneath the epithelium. Such distribution implies these nerve fibres to be involved in the regulation of the oviductal blood flow, non-vascular smooth myocyte tonus, transmission of sensory information and control of the epithelial secretion.