Horseradish peroxidase study of the location of extrinsic efferent and afferent neurons innervating the colon of dogs

The abdominal portion of the colon of 13 clinically normal dogs was divided into 5 regions (ascending, transverse, left colic flexure, proximal descending, and distal descending), and each region was injected with 30 mg of horseradish peroxidase (HRP). The injected colonic region, brain stem, L7-Cd1 portion of the spinal cord, sympathetic trunk ganglia, celiacomesenteric ganglia, caudal mesenteric ganglion, pelvic plexi, distal vagal (nodose) ganglia, and L1-Cd1 spinal ganglia were obtained at post-injection hour 48, sectioned, and processed by use of the tetramethylbenzidine method. The entire length of the colon was found to be under extrinsic influence of the parasympathetic nucleus of cranial nerve X (PX), with the largest average number of labeled cells resulting from injection of the ascending colon. It was also indicated that the entire colon is under extrinsic influence of the sacral portion of the spinal cord because the pelvic ganglia (second-order neurons) of the pelvic plexi contained labeled cells for all colonic regions. The largest average number of labeled cells in pelvic ganglia was seen after injection of the distal portion of the descending colon. Only after injection of the distal portion of the descending colon were labeled cells found in the S1-S3 portion of the spinal cord. Labeled cells in the PX, spinal cord, and pelvic ganglia were found bilaterally. Although the entire abdominal portion of the colon appears to be influenced by cranial and sacral parasympathetic preganglionic (via pelvic ganglia) neurons, the relative importance of the 2 areas seems to be reversed between the ascending colon and distal portion of the descending colon.(ABSTRACT TRUNCATED AT 250 WORDS)     

肾上腺与肾脏交感神经低级中枢及外周神经元定位研究进展

交感神经是植物性神经系统的重要组成部分。人们采用不同的研究方法,如染质溶解法、HRP法,对交感神经低级中枢位置即交感节前神经元定位和交感神经的外周神经元即交感节后神经元定位进行了广泛和深入的研究。结果表明,交感节前神经元位于胸腰段脊髓中间带区的不同核团内,支配不同器管的交感节前神经元在脊髓内有一定的分布节段;交感节后神经元位于外周植物性神经节内,如交感干神经节、腹腔神经节、肠系膜前神经节、主动脉肾神经节,支配不同器官的交感节后神经元在不同交感干神经节内有一定的分布节段。     

鸡胰腺交感传出神经元的分布研究

用霍乱毒素结合辣根过氧化物酶逆行追踪法对鸡胰腺的交感传出神经元的定位进行了研究.结果表明,鸡胰腺的交感传出神经元位于双侧的肾上腺神经节、内脏神经节和T3-T7交感干神经节,在交感干神经节中集中分布节段在T5交感干神经节.     

Diabetes mellitus-related morphoquantitative changes in the celiac ganglion neurons of the dog

Diabetes mellitus is the most common endocrine disturbance of domestic carnivores and can cause autonomic neurological disorders, although these are still poorly understood in veterinary medicine. There is little information available on the quantitative adaptation mechanisms of the sympathetic ganglia during diabetes mellitus in domestic mammals. By combining morphometric methods and NADPH-diaphorase staining (as a possible marker for nitric oxide producing neurons), type I diabetes mellitus-related morphoquantitative changes were investigated in the celiac ganglion neurons in dogs. Twelve left celiac ganglia from adult female German shepherd dogs were examined: six ganglia were from non-diabetic and six from diabetic subjects. Consistent hypertrophy of the ganglia was noted in diabetic animals with increase of 55% in length, 53% in width, and 61.5% in thickness. The ordinary microstructure of the ganglia was modified leading to an uneven distribution of the ganglionic units and a more evident distribution of axon fascicles. In contrast to non-diabetic dogs, there was a lack of NADPH-diaphorase perikarial labelling in the celiac ganglion neurons of diabetic animals. The morphometric study showed that both the neuronal and nuclear sizes were significantly larger in diabetic dogs (1.3 and 1.39 times, respectively). The profile density and area fraction of NADPH-diaphorase-reactive celiac ganglion neurons were significantly larger (1.35 and 1.48 times, respectively) in non-diabetic dogs compared to NADPH-diaphorase-non-reactive celiac ganglion neurons in diabetic dogs. Although this study suggests that diabetic neuropathy is associated with neuronal hypertrophy, controversy remains over the possibility of ongoing neuronal loss and the functional interrelationship between them. It is unclear whether neuronal hypertrophy could be a compensation mechanism for a putative neuronal loss during the diabetes mellitus.     

鸡胰腺交感传入神经元的定位研究

用霍乱毒素结合辣根过氧化物酶（CT—HRP）逆行追踪法对鸡胰腺的交感传入神经元的定位进行了研究。结果表明：支配鸡胰腺的交感传入神经元胞体分别位于T3-T7脊神经节，峰值位于T5、T6脊神经节。     

肾俞穴,后三里穴与子宫相关的神经基础

应用辣根过氧化物酶（ＨＲＰ）法和荧光素双标记法，研究了家兔子宫与肾俞穴、后三里穴的外周神经联系。结果显示，支配子宫与肾俞穴、后三里穴的初级传入神经元及交感节后神经元分布节段部分重叠，在腰１（Ｌ１）脊神经节有少量的荧光素双标记细胞，约占该节荧光素标记细胞总数的２．６％，表明Ｌ１脊神经节有一些初级传入神经元，其周围突分支分布于肾俞穴和子宫，提示子宫与肾俞穴除通过中枢途径联系外，还可通过外周突分支投射的感觉神经元直接相联系；在胸１２～荐３（Ｔ１２～Ｓ３）交感神经节出现大量分别支配子宫和后三里穴的标记细胞，这两种细胞紧密相邻，提示支配子宫和后三里穴的交感节后神经元之间，可能还存在直接或间接联系。上述结果为穴位－内脏相关机理提供了形态学证据。     

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.     

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.     

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.     

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.     

Analysis of the chemical coding of neurons in the intermediate thoracic ganglion of the pig

The pig has been widely used as a model in cardiovascular research. A unique feature of the porcine extrinsic sympathetic cardiac nerves is that they arise from intermediate ganglia in the thoracic cavity. The localization and pattern of distribution of nerve cell bodies and fibers containing tyrosine hydroxylase (TH), dopamine B-hydroxylase (DBH), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), methionine-enkephalin (MET) as well as calcitonin gene-related peptide (CGRP), substance P (SP) and pituitary adenylate cyclase-activating peptide (PACAP) was studied with immunohistochemistry. Almost all the neurons showed immunoreactivity to TH. Immunoreactivity to NPY, VIP, SOM, GAL, MET and PACAP was displayed by nerve cell bodies while nerve fibers exhibited immunoreactivity to all the neuropeptides studied. Therefore, it seems that the chemical coding of neurons and especially nerve fibers in the porcine intermediate ganglion share general similarities (with certain neurochemical variability), with porcine prevertebral ganglia (e.g., celiacomesenteric and caudal mesenteric ganglia).     

Location of autonomic cell bodies projecting to the extrathoracic smooth muscle of the canine trachea

A study of the origin of the sympathetic postganglionic axons innervating the trachealis muscle of the dog was carried out using horseradish peroxidase as a retrograde neuronal marker. It was demonstrated that this technique is not fruitful in adult dogs, but in 12 puppies demonstrated that these neurons arise from the cervicothoracic, middle cervical and cranial cervical ganglia. Contributions of nucleus ambiguus, nucleus of the vagus nerve and the distal ganglion of the vagus nerve to innervation of the trachealis muscle were also demonstrated.     

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.     

A Light and Electron Microscopic Study of Developing Bovine Sympathetic Ganglia

This study concentrated on the histogenesis of bovine sympathetic ganglia, especially the development of the capsule, the interaction of sympathoblasts and supporting cells, and the ultrastructural detail of developing sympathoblasts. For this purpose 1st thoracic ganglia from 19 embryos and foetuses were collected and separated into 4 groups. The methods of study involved light and electron microscopy. From the study it was concluded that for neuronal development a functional interaction was necessary with satellite and Schwann cells, that the capsule had a role in maintaining neuronal homeostasis, and that sympathetic neurons displayed a number of morphologically distinct forms before reaching maturity.     

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)     

A novel ultrasound-guided lateral quadratus lumborum block in dogs: a comparative cadaveric study of two approaches

ObjectiveTo describe an ultrasound-guided lateral quadratus lumborum (LQL) block technique and the spread characteristics of lidocaine–dye injected in the LQL plane using a transversal (LQL-T) or a longitudinal (LQL-L) approach.Study designExperimental anatomic study.AnimalsA total of eight canine cadavers.MethodsBilateral ultrasound-guided injections in the fascial plane lateral to the quadratus lumborum muscle and medial to the thoracolumbar fascia (LQL plane) with the needle directed at the first lumbar (L1) transverse process were performed using lidocaine–dye (0.3 mL kg⁻¹). Anatomical dissection determined the dye distribution, sympathetic trunk staining and number of spinal nerves stained circumferentially >1 cm.ResultsThe LQL fascial plane was ultrasonographically recognized in all cadavers and filled with lidocaine–dye in all eight cadavers with the LQL-T approach and in six with LQL-L. The injectate spread ventral to the lumbar transverse processes, around the quadratus lumborum muscle and dorsal to the transversalis fascia, affecting the ventral branches of the spinal nerves and the sympathetic trunk. A median (range) of 4 (3–5) and 3 (0–4) ventral branches of the thoracolumbar nerves were dyed with LQL-T and LQL-L approaches, respectively (p = 0.04). The most cranial nerve stained was the twelfth thoracic (T12) with the LQL-T approach and T13 with LQL-L, and the most caudal was L3 with both approaches. The incidence of sympathetic trunk staining was significantly higher using LQL-T (six injections) compared with LQL-L (one injection; p = 0.04). Dye was not observed in the lumbar plexus, epidural space or abdominal cavity.Conclusions and clinical relevanceUltrasound-guided LQL-T approach resulted in a more consistent spread toward the spinal nerves and sympathetic trunk compared with LQL-L approach. Further studies are necessary to assess the LQL block effectiveness and success rate in live dogs.     

支配家兔下颌淋巴结的神经来源

本实验用ＨＲＰ逆行追踪法对家兔下颌淋巴结的神经来源进行了研究。结果表明；在同侧三叉神经半月状神经节和Ｃ１－４脊神经出现标记神经原，以三叉神经经半月状神经的标记细胞最多，提示下颌淋巴结的初级感觉主要经三叉神经传入；在同侧的颈前节，星状神经节及Ｔ２－３椎旁节出现标记的交感后神经原，主要出现于颈前节的吻侧端。     

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.     

The spinal nerves that constitute the lumbosacral plexus and their distribution in the chinchilla

In this study, the spinal nerves that constitute the lumbosacral plexus (plexus lumbosacrales) (LSP) and its distribution in Chinchilla lanigera were investigated. Ten chinchillas (6 males and 4 females) were used in this research. The spinal nerves that constitute the LSP were dissected and the distribution of pelvic limb nerves originating from the plexus was examined. The iliohypogastric nerve arose from L1 and L2, giving rise to the cranial and caudal nerves, and the ilioinguinal nerve arose from L3. The other branch of L3 gave rise to the genitofemoral nerve and 1 branch from L4 gave rise to the lateral cutaneous femoral nerve. The trunk formed by the union of L4-5 divided into medial (femoral nerve) and lateral branches (obturator nerve). It was found that the LSP was formed by all the ventral branches of L4 at L6 and S1 at S3. At the caudal part of the plexus, a thick branch, the ischiadic plexus, was formed by contributions from L5-6 and S1. This root gave rise to the nerve branches which were disseminated to the posterior limb (cranial and caudal gluteal nerves, caudal cutaneous femoral nerve and ischiadic nerve). The ischiadic nerve divided into the caudal cutaneous surae, lateral cutaneous surae, common fibular and tibial nerve. The pudendal nerve arose from S1-2 and the other branch of S2 and S3 formed the rectal caudal nerve. The results showed that the origins and distribution of spinal nerves that constitute the LSP of chinchillas were similar to those of a few rodents and other mammals.     

Small-cell carcinoma of the lung resembling a brachial plexus tumour

A small-cell carcinoma of the lung was identified in a six-year-old female German shepherd dog with a history of chronic lameness of the left forelimb, Horner's syndrome and sensory deficits on the caudal portion of the left forelimb below the elbow. A mass, the exact location of which was difficult to ascertain, was identified during radiographic examination of the thorax. It was easily identified, using magnetic resonance imaging, as an apical tumour of the left lung with dorsal extension and involvement of paraspinal structures, such as spinal nerve roots C8 to T1 and the sympathetic trunk. Postmortem examination confirmed a mass in the left apical lobe of the lung, compatible with a diagnosis of small-cell carcinoma by histopathology and immunohistochemistry. This clinical presentation is similar to Pancoast syndrome described in humans.