Distribution of sympathetic and afferent neurones innervating the submandibular gland in the sheep

The distribution of sympathetic and sensory neurones innervating the submandibular gland (SMG) in sheep was studied using retrograde tracing technique. The retrograde tracer Fast Blue (FB) was unilaterally injected into the SMG in five juvenile male sheep under general anaesthesia. After a 4-week survival period, all the animals were reanaesthetized, perfused transcardially with 4% buffered paraformaldehyde and ganglia, which could be considered as a potential sources of sympathetic, and afferent innervation of the gland were bilaterally collected. The FB-labelled sympathetic neurones were found in the ipsilateral superior and middle cervical ganglion. Many labelled neurones were distributed in the ipsilateral jugular and nodose ganglia of the vagus nerve and smaller numbers of the nerve cells were also found in ipsilateral C1-C3 dorsal root ganglia (DRG). No labelled neurones were observed in the ipsilateral stellate ganglion, trigeminal ganglion, C4-C8 DRG and in all contralateral ganglia. The present study revealed that the majority of sympathetic neurones projecting to the sheep SMG are found in the superior cervical ganglion but some of them are also distributed in the middle cervical ganglion. Most of the afferent neurones are located in the jugular and nodose ganglia of vagus nerve but C1-C3 DRG also comprise some of these nerve cells.     

Origin and chemical coding of primary afferent neurones supplying the prostate of the dog

Retrograde tracing technique combined with the double-fluorescent immunohistochemistry were used to investigate the distribution and chemical coding of primary afferent neurones supplying the canine prostate. After the injection of Fast Blue (FB) into the prostatic tissue retrogradely-labelled (FB(+)) primary afferent neurones were localized in bilateral L(1)-Ca(1) dorsal root ganglia (DRG). Statistical analysis using anova test showed that there are two major sources of afferent prostate innervation. The vast majority of prostate-supplying primary afferent neurones were located in bilateral L(2)-L(4) DRG (56.9 +/- 0.6%). The second source of the afferent innervation of canine prostate were bilateral S(1)-Ca(1) DRG (40.6 +/- 1.0%). No statistically significant differences were found between average number of FB(+) neurones localized in the left and right DRG (49.5 +/- 1.7 and 50.5 +/- 1.7%, respectively). Immunohistochemistry revealed that FB(+) primary afferent neurones contain several neuropeptides in various combinations. In the prostate-supplying neurones of lumbar and sacro-caudal DRG the immunoreactivity to substance P (SP) and calcitonin gene-related peptide (CGRP) was found most frequently (50 +/- 3.7 and 37.3 +/- 1.9%, respectively). Both in the lumbar and sacro-caudal DRG, considerable population of FB(+) neurones immunoreactive neither to SP nor CGRP were also found (23 +/- 2.6 and 32.8 +/- 2.3%, respectively). In the lumbar DRG 10.7 +/- 1.1% of SP-immunoreactive FB(+) neurones also contained galanin (GAL). In 9.2 +/- 2.2% of the prostate-supplying primary afferent neurones located in the sacro-caudal DRG the co-localization of SP and GAL was also reported. Results of the retrograde tracing experiment demonstrated for the first time sources of afferent innervation of the canine prostate. Double immunohistochemistry revealed that many of the prostate-supplying primary afferent neurones express some of sensory neuropeptides which presumably may be involved in nociception and some pathological processes like inflammation or nerve injury.     

Localization of sympathetic, parasympathetic and sensory neurons innervating the heart of the Beijing duck by means of the retrograde transport of horseradish peroxidase

Sympathetic, parasympathetic and sensory neurons were labeled by injections of horseradish peroxidase into various regions of the heart in 33 Beijing ducks. Sympathetic postganglionic neurons innervating the heart were located in the paravertebral ganglia C15 (C16 is the last cervical segment in the duck) to T3, especially in the ganglion T1. The coronary sulcus and ventricle were more abundantly innervated by sympathetic neurons than the atrium. The left side of the heart was preferentially innervated by sympathetic postganglionic neurons in the left side of paravertebral ganglia but the right side of the heart were equally supplied from the right and left ganglia. Within the medulla oblongata, the number of labeled vagal preganglionic neurons in the nucleus ambiguus was much greater than that in the dorsal motor nucleus of the vagus nerve. Labeled neurons of the nucleus ambiguus were found in many ducks injected into the coronary sulcus. Cardiac sensory neurons were observed in the dorsal root ganglia C15 to T2 (highest in the ganglion T1) and in the nodose and jugular ganglia of the vagus nerve. These labeled neurons probably form the afferent and efferent limbs of cardiac reflexes and control circulation in the Beijing duck.     

Galanin-immunoreactive cells and their relation to calcitonin gene-related peptide-, substance P- and somatostatin-immunoreactive cells in rat lumbar dorsal root ganglia

We report upon the distribution of galanin-immunoreactive (GAL-IR) cells in the lumbar dorsal root ganglia (DRG) of the rat, and upon the distribution of GAL-IR cells, which also contain calcitonin gene-related peptide (CGRP)-, substance P (SP)- and somatostatin (SOM)-immunoreactivity. Neuropeptide-immunoreactive lumbar DRG cells were 55.8% for CGRP, 12.7% for SP, and 6.5% for GAL in lumbar DRG cells. There was no significant difference between the right and left DRGs (L1-L6) for any neuropeptide-immunoreactive cell (P < 0.01). In terms of size distribution, CGRP-immunoreactive cells were identified below 1500 microm2, and SP-, and GAL-IR cells below 600 microm2. Neuropeptide immunoreactive cells showed various immunoreactivities in the cytoplasm according to each neuropeptide. CGRP and SP immunoreactive cells were colocalized with GAL immunoreactive cells in the serial sections about 83.3 and 60% respectively, but SOM colocalizing with GAL-IR cells were not in evidence. The current results confirm and extend previous results, and show that neuropeptides can coexist in single sensory neurones of the rat DRG. In addition, our results demonstrate that the normal distribution of some neurotransmitters modulating sensory action in Wistar Kyoto rat, make this model more prone to develop neuropathic pain than Sprague-Dawley rat.     

Pathogenesis of experimental bovine spongiform encephalopathy (BSE): estimation of tissue infectivity according to incubation period

This paper reports the results of tissue infectivity assays of bovine spongiform encephalopathy (BSE) agent in orally exposed cattle at stages during the incubation period. Estimations of the titre of infectivity in central nervous system (CNS), certain peripheral nerve ganglia and distal ileum tissue were made according to time post exposure from the relationship between incubation period and dose for RIII mice and C57bl mice using data from titrations of brain material from cases of BSE. The rate of increase of infectivity in the bovine CNS was then estimated, taking into account these tissue infectivity titres, the variability of the brain titre of clinical field cases of BSE, and the probability density of the expected number of months before clinical onset of each infected bovine. The doubling time for CNS was shown to equal 1.2 months. The titre in the thoracic dorsal root ganglia (DRG) was, on average, approximately 1 log units less than CNS, and cervical DRG approximately 0.5 log less than thoracic DRG. The pattern of increase of infectivity in the distal ileum is that of an initial increase up to 14-18 months post exposure, followed by a decrease, which is likely to be highly variable between animals. These results will be informative for future risk assessments of BSE, especially in relation to reviewing current control measures.     

The cranial cervical ganglion and its branches in the yak (Bos grunniens)

The heads and necks of 10 yaks were dissected to study the shape, location, arrangement, and branches of the cranial cervical ganglion. The ganglion was a greyish fusiform structure, mean length 19.72 mm, width 7.65 mm and depth 4.55 mm, located on the rostrolateral surface of the m. longus capitis. Approximately 25% of the ganglion was covered by the tympanic bulla, the rest by the m. stylohyoideus. The branches of the cranial cervical ganglion included the internal and external carotid nerves, sympathetic trunk and the branches connecting with the glossopharyngeal, vagus and hypoglossal nerves. In one animal the right cranial cervical ganglia was a greyish pyramidal structure 10 mm long, 8 mm wide and 5 mm thick but the left ganglion was similar to those found in the other specimens examined.     

Distribution of neurons supplying the porcine mammary gland

The distribution of neurons projecting to the mammary gland was investigated by using the retrograde tracing method in juvenile pigs (n = 12). Fluorescent retrograde tracer Fast Blue (FB) was injected into the nipple (n = 3) or parenchyma (n = 3) of the second, right thoracic mamma or into the nipple (n = 3) and parenchyma (n = 3) of the last, right abdominal mamma. FB-positive (FB+) mammary gland-projecting neurons were found in some right dorsal root ganglia (DRG) and sympathetic chain ganglia (SChG) only. After injection of the tracer into the second, right thoracic mamma, FB+ neurons were observed in Th9-Th12 DRG but most of them were located in Th11 and Th12 ganglia. As concerns SChG, FB+ neurons were found in Th1-Th4, Th7-Th14 and L1-L4 ganglia. The vast majority of them were located in Th10 and Th11 SChG, which appeared to be the main sources of efferent innervation of this mamma. Neurons projecting to the last right abdominal mamma were found in L1-L3 DRG and L1-L4 SChG but most of them were located in L1-L2 ganglia and L1-L2 ganglia, respectively. This study for the first time has disclosed the localization of neurons supplying the mammary gland in larger domestic animal species, the pig, by using the retrograde tracing method.     

Preliminary studies on long distance, retrograde transport of horseradish peroxidase in equine peripheral nerves

As a prelude to studies on retrograde axonal transport of neurotoxin (ie, so-called suicide transport) as a means to prevent post neurectomy neuroma formation, preliminary studies were conducted with an innocuous enzymatic marker, horseradish peroxidase (HRP). The proximal stumps of resected medial and lateral palmar digital nerves in six ponies were injected via a tuberculin syringe and needle with 50 micron 1 of a 30 per cent solution of HRP in order to assess long distance retrograde axonal transport. The dorsal root ganglion of the cervical spinal enlargement (ie, C6, C7, C8, T1, T2) were removed at post injection intervals of two, four, six, eight, 10 and 12 days. These were sectioned serially and reacted by the tetramethylbenzidine method to demonstrate transported enzyme in the ganglionic cell bodies which give rise to sensory fibres of the palmar digital nerves. Enzyme, retrogradely transported over axon lengths of 115 cm, was first demonstrated in spinal ganglia four days after injections of the palmar digital nerves. The calculated transport velocity of 287 mm/day, although almost certainly an underestimate, greatly exceeded rates of 72 to 120 mm/day recorded previously with HRP in the peripheral nerves of small laboratory animals. The intensity of the HRP reaction product in ganglionic neurons was strong at four days and it remained unabated in ganglia examined at six, eight, 10 and 12 days post injection. The major sources of the sensory fibres of the palmar digital nerves appeared to be the ganglia of the C8 and T1 spinal segments which contained more than 90 per cent of all labelled neurons.(ABSTRACT TRUNCATED AT 250 WORDS)     

Macro anatomical investigations of the cranial cervical ganglion in domestic pig (Sus scrofa domesticus)

In this study, the left and right cranial cervical ganglia (ganglion cervicale craniale) of eight young (four male, four female) domestic pigs weighing around 70-80 kg were inspected macro anatomically. The cranial cervical ganglion (CCG) was found cranio-ventrally of the distal ganglion of the vagus nerve, medial of the jugular process extremity, ventral of the atlas, dorsal of the epiglottis base and medial of the common root (CR) established by the internal carotid and occipital arteries. The internal carotid nerve and jugular nerve ramified from the cranial part of CCG. The jugular nerve gave branches that merged with the vagus and glossopharyngeal nerves. Other nerve branches originating from the cranial part of the ganglion reached to the external carotid artery and CR. The internal carotid nerve varied among cadavers in number of branches (two to four). These branches did not travel along the side of the internal carotid artery. The central part of CCG gave thin nerve branches that reached to various anatomical structures including the first and second cervical nerves, wall of the pharynx, accessory nerve, hypoglossal nerve, vagus nerve, external carotid artery and CR. The caudal part of CCG gave nerve branches that merged with the vagus, cranial laryngeal nerves, and common carotid artery. The external carotid nerves, which were two or three in number, also originated from the caudal part of CCG. In conclusion, the nerves ramifying from CCG of the pig varied in number among cadavers. Compared with literature raised in other species, there are also differences in number of nerve branches and course pattern of these nerves.     

Location of sympathetic postganglionic and sensory neurons innervating the testis in the male chicken

Sympathetic postganglionic and sensory neurons were labelled by injections of horseradish peroxidase into the testis of the male chicken. The total number of labelled neurons in the paravertebral, prevertebral, dorsal root and nodose ganglia was 943 on average for five chickens. Sympathetic postganglionic neurons were located in the paravertebral ganglia T3-LS3 (10% of the total number of labelled neurons), especially in T6 and T7, and in the prevertebral ganglia adjacent to the adrenal glands and aorta (19%). They were found almost ipsilaterally. No labelled neurons were observed in the dorsal motor nucleus of the vagus. Sensory neurons were found bilaterally in the dorsal root ganglia T2-LS3 (71%), especially in T5 to T7. Over a quarter of labelled sensory neurons were located in the contralateral dorsal root ganglia. In the nodose ganglia, only a few labelled sensory neurons were observed (much less than 1%). These results indicate that, unlike the ovary, the testis of the chicken tends to be innervated by ipsilaterally located sympathetic postganglionic and sensory neurons, with the sensory neurons being more numerous than the sympathetic postganglionic neurons.     

Fluoroscopy-guided approach to the lumbar dorsal root ganglion in dogs: cadaver study in Beagles

ObjectiveTo evaluate an approach to the canine lumbar dorsal root ganglion (DRG), a significant contributor to the pain pathway, using new methylene blue staining.Study designProspective randomized study.AnimalsA total of three Beagle dog cadavers weighing 10.4 ± 0.7 kg (mean ± standard deviation).MethodsBilateral third to fifth lumbar DRG approaches were performed in three dog cadavers positioned in sternal recumbency. The mammillary process was palpated, and a 22 gauge spinal needle was inserted through the skin 1 cm lateral to the process and directed towards the median plane at a 45° angle to the dorsal plane. The needle was advanced along the transverse plane until touching bone, or a popping sensation was detected. Under fluoroscopic guidance, the position of the needle tip was adjusted to be in the cranioventral part of the intervertebral foramen. The location of the needle was confirmed by demarcation of the nerve roots after iohexol (0.1 mL) injection. For evaluation of the DRG approach, new methylene blue (0.1 mL) was injected. Subsequently, anatomical dissection of the area was performed. The DRG staining was scored as follows: 0, no staining; 1, partial (<50%); 2, partial (≥50%); and 3, complete staining. Comparisons among the staining scores of the third to fifth DRG were assessed with the Friedman test.ResultsStaining score 3 was achieved in 14 of 18 (77.8%) sites. Staining scores 2, 1 and 0 were identified at two, one and one of the 18 sites, respectively. No significant difference was noted in the staining scores among the third to fifth DRGs (p = 0.78).Conclusions and clinical relevanceThe technique used for DRG injections achieved adequate DRG staining, supporting use of the fluoroscopy-guided approach to the canine lumbar DRG.     

Gross anatomy of the accessory nerve and vagus nerve ofthe head and cranial neck region in the Bactrian camel

Seven heads and necks of Bactrian camels were dissected to investigate the origin, course, branches anddistribution of the accessory nerve and vagus nerve in the cranial cervical region. The spinal root and external branch of the accessory nerve were not present, but there was a delicate communicating branch between the dorsal root of the first cervical nerve and the root of the vagus nerve. The sternocephalic muscle was innervated by the second cervical nerve while the brachiocephalic and trapezius muscles were supplied by the sixth and seventh cervical nerves. In the head and cranial cervical region of the Bactrian camel the vagus nerve gave oft the auricular branch, pharyngeal branch, cranial laryngeal nerve, a common trunk to the larynx, oesophagus and trachea, and some communicating branches connecting with the glossopharyngeal, hypoglossal, first cervical nerves and the cranial cervical ganglion.     

Sources of Sensory Innervation of the Hip Joint Capsule in the Rabbit – A Retrograde Tracing Study

The aim of the study was to investigate the sensory innervation of the hip joint capsule in the rabbit. Individual animals were injected with retrograde fluorescent tracer Fast Blue (FB) into the lateral aspect of the left hip joint capsule (group LAT, n = 5) or into the medial aspect of the hip joint capsule (group MED, n = 5), respectively. FB‐positive (FB+) neurons were found within ipsilateral lumbar (L) and sacral (S) dorsal root ganglia (DRG) from L7 to S2 (group LAT) and from L6 to S4 (group MED). They were round or oval in shape with a diameter of 20–90 μm. The neurons were evenly distributed throughout the ganglia. The average number of FB+ neurons was 16 ± 2.8 and 27.6 ± 3.5 in rabbits from LAT and MED, respectively. The largest average number of FB+ neurons in animals of group LAT was found within the S1 DRG (8 ± 1.7), while S2 ganglion contained the smallest number of the neurons (3.6 ± 1). In the L7 DRG, the average number of FB+ neurons was 6.2 ± 1.6. In rabbits of MED group, the largest number of FB+ neurons was found within the S1 DRG (13.4 ± 4), while the smallest one was found within the S3 ganglion (1.4 ± 0.4). In L6, L7, S2 and S4 ganglia, the number of retrogradely labelled neurons amounted to 1.6 ± 0.5, 4 ± 1.5, 4.4 ± 1.5 and 2.8 ± 1.7, respectively. The data obtained can be very useful for further investigations regarding the efficacy of denervation in the therapy of hip joint disorders in rabbits.     

Immunofluorescence studies on the pathogenesis of hemagglutinating encephalomyelitis virus infection in pigs after oronasal inoculation

Hemagglutinating encephalomyelitis virus (HEV; also designated vomiting and wasting disease virus) was inoculated oronasally in 14 colostrum-deprived pigs at the day of birth. Anorexia and vomition were seen after 4 days. Pigs were killed at different times after inoculation, and the results of the examination by immunofluorescent antibody technique revealed that the epithelial cells of nasal mucosa, tonsils, lungs, and small intestine served as sites of primary viral replication. After the local replication near the sites of entry, the virus spread via peripheral nervous system to the CNS. During the incubation period, viral antigens were detected in the trigeminal ganglion,the inferior vagal ganglion, the superior cervical ganglion, the intestinal nervous plexuses, the solar ganglion, and the dorsal root ganglia of the lower thoracic region. In the brain stem, the infection started in the trigeminal and vagal sensory nuclei and spread to other nuclei and to the rostral part of the brain stem. In later stages of the infection, viral spread into the cerebrum, cerebellum, and spinal cord was sometimes also observed. Viral replication in nervous plexuses of the stomach was not present during the incubation period, but was detected in all except 1 of the pigs that were ill when killed. The question whether the vomition is induced centrally by viral replication in the brain stem or is due to viral replication in peripheral nervous tissues remains unanswered.     

Neurochemical properties of aquaporin 1-expressing sensory neurons from the ovine trigeminal ganglion

Aims of the present study were to investigate the distribution and morphology of aquaporin 1-immunoreactive (AQP1-IR) neurons in the sensory ganglia of the sheep. Double immunohistochemical staining was applied to figure out whether substance P (SP), calcitonin gene-related peptide (CGRP) and galanin are present in AQP1-bearing primary afferent neurons. The expression of AQP1 was present only in trigeminal ganglion, whereas in nodose ganglion, jugular ganglion as well as C(1) -C(7) dorsal root ganglia no presence of AQP1 was found. In trigeminal ganglion, 15.4 ± 2.3% of Hu C/D-IR neurons (pan-neuronal marker) showed the presence of AQP1. The vast majority of AQP1-IR trigeminal sensory neurons (approximately 69.6 ± 3.3%, n = 5) were classified as middle in size, 28.6 ± 3.0% of AQP1-IR neurons were small and only 1.8 ± 0.6% of AQP1-positive neurons were large in size. Amongst the population of AQP1-IR trigeminal neurons as many as 58.5 ± 3.9% were immunopositive to SP, 30.7 ± 2.3% showed the presence of CGRP and 10.9 ± 0.2% coexpressed galanin. In trigeminal ganglion, SP-IR as well as CGRP-IR (but not galanin-IR) nerve fibres were found in close neighbourhood of AQP1-IR neurons. It is concluded that AQP1 is present in certain neuronal subsets of the ovine trigeminal ganglion; however, the exact role of this water channel has to be elucidated.     

Lymphocytic ganglioneuritis secondary to intervertebral disc extrusion in a dog

This study presents a case of lymphocytic ganglioneuritis in a dog secondary to intervertebral disc extrusion that mimicked a peripheral nerve sheath tumour on magnetic resonance imaging. A four‐year‐old spayed female dachshund with lumbar pain was imaged via magnetic resonance. A tubular, space‐occupying, contrast‐enhancing lesion was noted in the right intervertebral foramen at L6 to L7. This was presumed to represent focal enlargement of the right sixth lumbar spinal nerve. A right‐sided haemilaminectomy was performed at L6 to L7 and material that grossly resembled extruded nucleus pulposus was removed. The right L6 dorsal root ganglion, dorsal nerve root and proximal spinal nerve were severely enlarged and a partial thickness biopsy was collected from the dorsal root ganglion. Results of histopathological examination of the submitted tissue samples were consistent with extruded disc material and lymphocytic ganglioneuritis. To the author's knowledge, this is the first published report of lymphocytic ganglioneuritis secondary to intervertebral disc disease in a dog.     

Localization of Vesicular Glutamate Transporter 2 mRNA in the Dorsal Root Ganglion of the Pigeon (Columba Livia)

Our previous study showed localization of glutamate receptor 1 (GluR1) mRNA in neurons of the pigeon spinal cord, suggesting glutamatergic input from intrinsic and extrinsic origins. The present study examined localization of vesicular glutamate transporter 2 (VGLUT2) mRNA to confirm an extrinsic origin of glutamatergic neurons in the dorsal root ganglion (DRG). GluR1 and GluR2 mRNAs were examined in DRG and spinal cord to seek projection regions from VGLUT2 mRNA‐expressing neurons. VGLUT2 mRNA was expressed in most DRG neurons and labelling intensity varied from weakly to intensely. Intense VGLUT2 mRNA expression was mainly seen in medium to large neurons. GluR1 and GluR2 mRNAs were expressed in the dorsal horn and GluR2 mRNA signal was also seen in the marginal nucleus. The results suggest that the pigeon DRG has an extrinsic glutamatergic origin that project to the dorsal horn and marginal nucleus of the spinal cord.     

Ultrasonographic anatomy of the bovine eye.

The purposes of the study were to describe the ultrasonographic appearance and measurements of the normal bovine eye, to compare the measurements to those reported previously for cadaveric eyes and to describe differences between ocular dimensions of Holstein Friesian and Jersey cattle. Sixty transpalpebral ocular ultrasonographic examinations were performed on 30 adult Holstein Friesian cows, and 16 examinations were performed on 8 adult Jersey cows. Transpalpebral ultrasonographic images were obtained with a 10 MHz linear transducer in both horizontal and vertical imaging planes. The ultrasonographic appearance of structures within the bovine eye is similar to that in other species, although the ciliary artery was frequently identified, appearing as a 0.33 +/- 0.04 cm diameter hypoechoic area. The axial length of the globe was significantly greater in Holstein Friesian cattle (3.46 +/- 0.09 cm) compared with Jersey cattle (3.27 +/- 0.19 cm; P = 0.001), although the vitreous depth was smaller in Holstein Friesian cattle (1.46 +/- 0.09 cm) (P = 0.0009). The anterioposterior depth of the lens was significantly greater in Jersey cattle (1.92 +/- 0.11 cm) and the cornea was thinner in Jersey cattle (0.17 +/- 0.02 cm). The appearance and ocular distances for live animals were similar to those reported previously for cadaveric specimens. The knowledge of normal ocular dimensions facilitates the use of ultrasonography in the evaluation of ocular disease in cattle.     

Origin, course and distribution of the hypoglossal nerve in the New Zealand rabbit (Oryctolagus cuniculus L)

With 8 figures SUMMARY: This study aimed at revealing the origin, course and distribution of the hypoglossal nerve in 20 adult male New Zealand rabbits. In all the animals dissected, the hypoglossal nerve arose from the ventrolateral side of the medulla oblongata with two main roots and gave off a descending branch to the ansa cervicalis before reaching the division of the common carotid artery. This branch was not seen on the right side of only one case. At the lateral aspect of the hyoglossus muscle, the nerve then divided into the lateral and medial main branches, sent branches to the styloglossus, hyoglossus, genioglossus and geniohyoideus muscles and terminated in the intrinsic tongue muscles. A communicating branch was observed between the hypoglossal and accessory nerves in the right side of one animal and between the hypoglossal nerve and the ganglion nodosum in the right retropharyngeal area of another animal. An additional branch was observed innervating the stylohyoideus muscle in one animal only. A lateral lingual-hypoglossal communication was also seen between the lateral branch of the hypoglossal nerve and terminal branches of the lingual nerve.     

Immunohistochemical characteristics of neurons supplying the porcine bulbourethral gland

In the male pig, the bulbourethral gland (BG) is a particulary well developed accessory genital gland (AGG) which produces complex secretion contributing to the fluid component of semen. The secretory and motor function of AGGs is thought to be under the autonomic nervous system control. Although relatively much is known about the innervation of the prostate gland and, to a lesser degree, of the seminal vesicle, the paucity of data dealing with the innervation of BG is striking. Therefore, combined retrograde tracing and double-labelling immunofluorescence have been used to investigate the distribution and immunohistochemical properties of autonomic and primary afferent neurons projecting to this gland in the pig. BG-projecting neurons were found in some ipsilateral (I) and contralateral (C) sympathetic chain ganglia (SChG), the caudal mesenteric ganglion (CaMG), pelvic ganglia (PG) and some dorsal root ganglia (DRG). Immunohistochemistry revealed that the vast majority of CaMG and SChG BG-projecting neurons contained tyrosine hydroxylase (TH) and dopaminebeta-hydroxylase (DbetaH), and some neuropeptides including neuropeptide Y (NPY), somatostatin (SOM) and galanin (GAL). Three subpopulations of PG neurons supplying BG could be distinguished: 1) cholinergic neurons [vesicular acetylcholine transporter (VAChT)-positive] which also contained vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), SOM and NPY, 2) adrenergic neurons (TH-positive) which also stained for NPY, GAL or leu5-enkephalin (LEU), and 3) non-adrenergic, non-cholinergic neurons (NANC). DRG BG-projecting neurons contained mostly substance P (SP) and/or calcitonin gene-related peptide (CGRP) which sometimes colocalized with GAL. The possible functional significance of the substances found within the neurons is discussed.