Distribution of cholinergic and adrenergic nerve fibers in the lacrimal glands of dogs

The distribution of postganglionic autonomic nerve fibers in the lacrimal gland and gland of the third eyelid of dogs was studied by use of histochemical techniques. Adrenergic nerve distribution was identified by use of the sucrose-potassium phosphate-glyoxylic acid technique. A loose network of adrenergic nerves was found throughout the interstitium around acini and blood vessels and in vessel walls. Acetylcholinesterase staining was used to identify cholinergic nerve fibers. A cholinergic distribution pattern was found, although the cholinergic innervation appeared more dense than the adrenergic. In the gland of the third eyelid, mucus-secreting lobules and lipid-secreting lobules appeared to be equally innervated by parasympathetic fibers. These lobules could not be differentiated when the sucrose-potassium phosphate-glyoxylic acid technique was used. The techniques used in this study could not demonstrate whether direct contact was made by either cholinergic or adrenergic nerve fibers with secretory or myoepithelial cells. The presence of both nerve fiber types around acini suggests an interrelationship between the sympathetic and parasympathetic nervous system in lacrimal gland secretion in dogs.     

Immunohistochemical study on the expressions of neuropeptides in the superficial and deep gland of the third eyelid of pigs

The superficial gland of the third eyelid (SGTE) and deep gland of the third eyelid (DGTE) are classified as accessory lacrimal glands. The aim of the present study was to immunohistochemically investigate the expression of vasoactive intestinal peptide (VIP), dopamine beta-hydroxylase (DβH), substance P (SP), galanin, neuropeptide Y (NPY), pituitary adenylate cyclase-activating peptide (PACAP), somatostatin and calcitonin gene-related peptide (CGRP) in the porcine SGTE and DGTE. We demonstrated the distribution patterns of VIP, DβH, SP, NPY and galanin in the nerve fibres in the SGTE and DGTE. None of somatostatin-, PACAP- and CGRP-immunoreactive (IR) nerve fibres were found in the SGTE and DGTE. The majority of VIP- and DβH-IR nerves fibres were found near to glandular acini, tubules, secretory ducts and blood vessels in the SGTE and DGTE. VIP-IR nerve fibres were found in external connective tissue in SGTE and DGTE and only in interlobular connective tissue in the SGTE. DβH-IR nerve fibres were found in interlobular and external connective tissue in the DGTE but not in the SGTE. Single galanin-, SP- and NPY-IR nerve fibres were observed in close proximity to acini and tubules in the SGTE and DGTE. Single galanin-, SP-, NPY-IR nerve fibres were found in close proximity to the secretory ducts in the DGTE, however only SP-IR nerve fibres were found near to the secretory ducts in SGTE. In conclusion, our research aims to highlight some aspects of SGTE and DGTE innervation in pigs and may also be a source of basic knowledge for further studies.     

The distribution of nerve fibres immunoreactive for vasoactive intestinal peptide, calcitonin gene-related peptide, substance P and dopamine beta-hydroxylase in the normal equine larynx

The autonomic innervation of the mammalian respiratory system is complex, and involves a wide variety of peptide and non-peptide neurotransmitters which will have an important role in normal laryngeal function and the response to disease. This innervation has been partially described in the horse airway and lung, but there is no information on the equine larynx. This paper describes the expression and distribution of nerve fibres immunoreactive for vasoactive intestinal peptide (VIP), calcitonin gene-related peptide (CGRP), substance P (SP) and the adrenergic enzymatic marker dopamine beta-hydroxylase (DBetaH) in the mucosa of the equine larynx. The overall relative density of nerve fibres immunoreactive for the different antigens was VIP>CGRP>SP>DBetaH. There were differences in the distribution of nerve fibre types, although each antigen was found in nerve fibres adjacent to blood vessels and mucous glands. VIP -like immunoreactivity (VIP -Li) was particularly extensive in association with mucous glands. SP - and CGRP -like immunoreactivity (SP -Li, CGRP -Li) were also seen close to the epithelium, with occasional nerve fibres coursing beneath and between the epithelial cells. Fragments of SP -Li and CGRP -Li fibres were also present in large nerve fibre bundles and ganglionic cell clusters, but not in the neurons themselves. The density of nerve fibres immunoreactive for DBetaH was very low and restricted to blood vessels and mucous glands. There was marked variation in the density of nerve fibres at the different sites, with the greatest density, particularly for VIP, over the arytenoid cartilage. Immunoreactive nerve fibres were less plentiful over the epiglottis, and the density of all types of nerve fibres was low over the cricoid cartilage. Overall VIP -Li nerve fibres were the most plentiful.     

山羊子宫内胆碱能神经分布及妊娠时的变化

采用乙酰胆碱酯酶组化法，研究了山羊子宫内胆碱能神经的分布，结果，山羊子宫颈是经较丰富，在浆膜和肌层内有神经束伴血管而行并分支分布于血管壁，在粘膜及其皱褶上皮下，粘液腺上皮有神经丛分布，妊娠时子宫颈部的神经分布与未妊娠时相比无明显变化，子宫角部神经密度均低于子宫颈部，其内环行肌层中及其与内膜交界处神经密度略高，神经束伴血管而行并分支分布于血管壁，在子宫腺上皮下及内膜上皮下无神经分布，妊夺时作胎盘内无神经分布外，仍有神经束伴血管而行交分支分布于血管壁，在分布于血管壁的神经纤维减少，结果提示，胆碱能神经主要支配山羊子宫内血管壁及颈部粘液腺上皮和粘膜上皮，妊 时子宫颈部胆碱能神经无明显变化，而子宫角内支配血管壁的胆碱能神经纤维减少。     

Anatomy and Cholinergic Innervation of the Sinus paranalis in Dogs

Conventional methods have been used to study the general configuration of the wall of the Sinus paranalis in dogs. Apocrine tubular glands, elastic fibres and smooth-muscle fibres are the more significant elements of the wall, together with the epithelium and connective tissue. By means of the immunohistochemical method described in this paper, the results provide histochemical evidence for the presence of an AChE-positive reaction, and supposedly cholinergic nerve fibres in the wall of the dog anal sacs, mainly associated with subepithelial smooth muscle, vessels, and glands.     

Has Active Immunization Against Gonadotrophin‐Releasing Hormone any Effect on Testis Innervation in the Pig? – An Immunohistochemical Study

The innervation of porcine testes was studied in intact animals and in boars undergoing active immunization against gonadotrophin-releasing hormone (GnRH) by means of immunohistochemistry using antibodies to tyrosine hydroxylase (TH), dopamine beta-hydroxylase (D beta H), vasoactive intestinal polypolypeptide (VIP), neuropeptide Y (NPY), synaptosome-associated protein of 25 kDa (SNAP-25) and protein gene product 9.5 (PGP 9.5). Moreover, the distribution of luteinizing hormone (LH) receptors in clusters of Leydig cells was also investigated. To identify these cells easily, either the NADPH-diaphorase histochemical technique or the Mayer counter-staining procedure was applied. Differences in the distribution pattern and relative density of particular subsets of intratesticular nerve fibres were observed in immunized boars as compared to those found in the intact animals. In the testes of non-treated animals, only single TH-immunoreactive (TH-IR) nerve fibres were observed. However, many D beta H-IR nerve terminals surrounded blood vessels in the tunica albuginea and parenchyma. Very scarce VIP-IR nerves occurred only in the tunica albuginea, mainly in close vicinity to blood vessels. Immunoreactivity to NPY occurred in single nerve fibres. Immunoreactivity to SNAP-25 and PGP 9.5 was found in single nerve fibres distributed mainly in the tunica albuginea. The interstitial cells were heavily stained for LH-receptors and NADPH-diaphorase. In the testes of immunized animals, only single TH-IR nerve fibres, scattered mainly in the tunica albuginea, were observed. Some TH-IR nerve terminals were also encountered in the parenchyma of the organ, where they were always associated with blood vessels. D beta H-IR nerve fibres formed a dense network distributed throughout the testis in association with the capsule, vasculature and interstitium. Some fibres were observed to run between seminiferous tubules. VIP-IR nerve fibres were located in the neighbourhood of blood vessels in the tunica albuginea and parenchyma. Only single VIP-IR nerves were found between seminiferous tubules. Numerous NPY-IR nerve fibres occurred in the tunica albuginea and parenchyma of the organ. SNAP-25-IR and PGP 9.5-IR nerve terminals formed a dense network distributed throughout the testis and many fibres were observed between seminiferous tubules. Interstitial cells were very weakly stained for LH receptors or NADPH-diaphorase.     

Innervation of the circumvallate papilla of the cat tongue

Immunohistochemically, the distribution of S-100 protein and acetylated tubulin-positive nerve fibres was studied in the circumvallate papilla and its taste buds (TB) in the adult cat. The immunostaining for acetylated tubulin demonstrated an extensive innervation of the circumvallate papilla. Vegetative ganglionic cells were found in the central area of papilla, whereas fine nerve fibres were concentrated under epithelium. Individual positive axons were seen in relation to TB. Nerve fibres enter the TB and branch out among the supporting and sensory cells. Some nerve fibres reach the apical surface of the TB, including the taste pore. Nerve fibres positive for S-100 protein were observed as dense nerve plexus located in the core of the papilla. Satellite cells localized inside the papilla were seen to surround the ganglionic cells. Bands of fine nerve fibres were present under lining epithelium, mainly at the base of the TB. A weak reaction was displayed by taste bud cells and surrounding epithelial cells as well as by the epithelial cells of the papillary side of the moat. A dense network of the nerve fibres was present among the glandular acini and surrounding the ducts of the serous Ebner's glands.     

Immunohistochemical study of the pre- and postnatal innervation of the dog lower urinary tract: morphological aspects at the basis of the consolidation of the micturition reflex

Immunohistochemical studies were performed on male and female bladder and urethra collected from 4 adults dogs and 10 foetal specimens with crown-rump length from 53 to 155 mm (medium-sized breeds, presumptive 38 days of gestation to term). A panel of antisera was tested, including PGP 9.5 to describe the general intramural innervation, ChAT and TH to depict the cholinergic and nor-adrenergic components and NOS1, CGRP, SP, NPY, VIP, SOM, GAL, 5-HT to investigate the possible nitrergic, peptidergic and aminergic ones. A rich cholinergic innervation was present in adult bladder and urethra, along with a lesser number of adrenergic nerves and a small number of nitrergic ones. Either bladder or urethra received numerous CGRP-, SP-, NPY-, VIP-containing nerve fibres which were distributed throughout the muscle layers. All over the lower urinary tract strong to weak ChAT-, CGRP-, SP- and NPY-immunoreactivity was detected in intramural ganglia, in peripheral nerve bundles and around blood vessels. 5-HT-immunoreactive endocrine cells were present in the urethral epithelium. Early foetal organs were supplied only by cholinergic nerve fibres. Few NOS-, CGRP- and SP-ergic components appeared at the end of pregnancy. It can be guessed that sensory mediators such as CGRP and SP increase in postnatal ages while other neuropeptides, such as NPY and VIP, appear only after birth, as the urinary reflex consolidates.     

Tyrosine hydroxylase- and neuropeptides-immunoreactive nerves in canine trachea

OBJECTIVE: To determine distribution of catecholaminergic and peptidergic nerve fibers in canine tracheas by use of immunohistochemistry. SAMPLE POPULATION: 10 tracheas collected from healthy adult dogs after euthanasia. PROCEDURE: Structure of the nerve network and distribution of tyrosine hydroxylase (TH)- and 6 types of neuropeptide-containing nerves in canine tracheas were immunohistochemically studied, using neurochemical markers. RESULTS: Intraepithelial free nerve endings with immunoreactivity for calcitonin gene-related peptide (CGRP) and substance P (SP) were observed.Tyrosine hydroxylase-, SP-, vasoactive intestinal peptide (VIP)-, and galanin (GAL)-immunoreactive nerve fibers were observed within and around the submucosal seromucous gland. In the smooth muscle layer, numerous TH- and GAL-immunoreactive nerve fibers, a moderate number of VIP- and neuropeptide Y (NPY)-immunoreactive nerve fibers, and a few SP- and methionine enkephalin (ENK)-immunoreactive nerve fibers were observed. Numerous nerve cell bodies with VIP and GAL immunoreactivity and a few with SP ENK, and NPY immunoreactivity were observed. Many TH-immunoreactive fibers were arranged in a meshwork around blood vessels. Nerves with CGRP-, SP-, VIP-, GAL-, ENK-, and NPY-immunoreactivity were also observed around blood vessels. CONCLUSIONS: Complex innervation, including catecholamine- and neuropeptide-containing nerves, which may be related to regulation of muscle contraction and glandular secretion, are found in canine tracheas.     

Nerve fibres in the parathyroid gland of the golden hamster (Mesocricetus auratus): immunohistochemical and ultrastructural investigations

We investigated the morphology and the distribution of the nerve fibres in golden hamster (Mesocricetus auratus) parathyroid glands using antibodies to calcitonin gene-related peptide (CGRP) and substance P, and electron microscopy. CGRP-immunoreactive nerve fibres were densely distributed in the interstitial tissues and the capsules of the hamster parathyroid glands. Some nerve fibres were detected in close proximity of the parathyroid chief cells. The distribution pattern for substance P-immunoreactive nerve fibres was roughly the same as for CGRP-immunoreactive fibres. Ultrastructurally, we found numerous nerve fibres joining the blood vessels. Axon bundles were located adjacent to the smooth muscle cells of the arterioles. The axons formed structurally specialized neuromuscular junctions with the vascular smooth muscle cells. Some axons were in close vicinity to the parathyroid chief cells. These findings indicate that the hamster parathyroid gland contain CGRP and substance P, which may regulate the blood flow and the secretory activity of the gland.     

Immunoreactivities to protein gene product 9.5, neurofilament protein and neuron specific enolase in nerves in the oviduct of the sexually immature ostrich, Struthio camelus

The present study investigated the distribution of nerves in the oviduct of the sexually immature ostrich. The presence of protein gene product 9.5, neurofilament protein and neuron specific enolase nerve fibres were demonstrated in the infundibulum, magnum, isthmus, shell gland and vagina. Nerve fibres containing protein gene product 9.5, neuron specific enolase and neurofilament protein were particularly numerous in the tunica muscularis and intermuscular connective tissue areas of the shell gland and vagina. The presence of a large number of nerves in these oviductal regions is probably important in the coordination of muscle contraction. An interesting finding of the study was the presence of protein gene product 9.5 and neuron specific enolase immunopositive nerve fibres in the walls of blood vessels. It is likely that these nerves are autonomicin nature and play a role in the regulation of blood flow. This study has shown the presence of an extensive neural network in the oviduct of the ostrich. In addition, the results of the investigation have shown that the neuronal markers protein gene product 9.5, neurofilament protein and neuron specific enolase can be used to demonstate nerve fibres in the ostrich.     

An Immunohistochemical Survey of Catecholamine-synthesizing Enzyme-immunoreactive Nerves and Endocrine Cells in the Bovine Pancreas

The distribution of catecholamine-synthesizing enzyme-immunoreactive nerves and endocrine cells in the pancreas of the calf and cow was studied immunohistochemically using antisera against tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). TH- and DBH-immunoreactive nerve fibres were abundant both within and around the islet of Langerhans. A few TH- and DBH-immunoreactive nerve fibres were seen around the large islets characteristic of calf pancreas, but the majority of cells in the large islets, and some in islets of Langerhans, showed TH immunoreactivity. In the exocrine pancreas, both TH- and DBH-immunoreactive nerve fibres were distributed randomly among the acini, with the DBH-immunoreactive fibres being more numerous. Abundant TH- and DBH-immunoreactive nerve fibres were seen in close association with blood vessels and in the connective tissue around the interlobular duct. Immunoreactivity for both enzymes was also observed in the nerve cell bodies and fibres of the intrapancreatic ganglia. The findings suggest an important role for catecholamines in the regulation of bovine pancreatic function.     

Organisation of Autonomic Nervous Structures in the Small Intestine of Chinchilla (Chinchilla laniger,Molina)

Using histochemical, histological and immunocytochemical methods, organisation of the autonomic nerve structures in small intestine of chinchilla was investigated. Myenteric plexus was localised between circular and longitudinal layers of the smooth muscles. Forming network nodes, the small autonomic, cholinergic ganglia were linked with the bundles of nerve fibres. Adrenergic structures were visible as specific varicose, rosary‐like fibres forming bundles of parallel fibres connecting network nodes. Structures of the submucosal plexus formed a finer network than those of the myenteric plexus. Moreover, in ‘whole‐mount’ specimens, fibres forming thick perivascular plexuses were also observed. Immunocytochemical studies confirmed the cholinergic and adrenergic character of the investigated structures. VAChT‐positive neurones were found only in myenteric plexus, and numerous VAChT‐positive and DBH‐positive fibres were found in both plexuses.     

Noradrenergic and cholinergic innervation of the accessory sexual glands in male sheep

Immunohistochemistry was applied to determine the distribution patterns of nerve fibres containing tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH), neuropeptide Y (NPY), vasoactive intestinal peptide (VIP) and vesicular acetylcholine transporter (VAChT) in the prostate, seminal vesicle (SV) and bulbourethral glands (BU) of male sheep. In all organs studied, cholinergic innervation was more developed than noradrenergic innervation. Numerous VAChT-immunoreactive (IR) nerve fibres were found in the muscular layer and mucosa of the SV and BU as well as in the prostate. A similar abundance of noradrenergic nerve fibres (showing immunoreactivity both to TH and DbetaH) was also found in both layers of the SV and BU (but not in the prostate). In the prostate a moderate density of VIP-IR nerve fibres was present but only very scarce NPY-IR nerve fibres were shown. All the studied accessory sexual glands (ASG) of male sheep contained VIP-IR nerve fibres in a similar frequency. Double immunohistochemistry revealed that the vast majority of noradrenergic nerve fibres also contained NPY. None of the noradrenergic nerve fibres showed the presence of VAChT or VIP. The possible functional significance of these findings is discussed.     

The Lymph Drainage Pattern of the Mammary Glands in the Cat: A Lymphographic and Computerized Tomography Lymphographic Study

Seventy-three clinically normal, lactating cats were used to investigate the lymph drainage of 73 mammary glands. In 50 cats of the first group, the number of lymphatic vessels emerging from the examined mammary gland, their course and the lymph nodes into which they are drained were studied by indirect lymphography (IL) after intramammary injection of an oily contrast medium. In 23 cats of the second group, the lymph drainage of the mammary glands was studied by computerized tomography indirect lymphography (CT-IL) after intramammary injection of a water soluble contrast medium. The following day, the lymph drainage of the mammary gland examined by CT-IL was studied by IL, as it was described in the first group, for comparison purposes. The main conclusions drawn after this study were as follows: lymph drains from the first and second mammary glands with one or rarely two or three lymphatic vessels to the accessory axillary lymph nodes. Lymph drains from the third mammary gland with one or two and rarely three lymphatic vessels usually to the accessory inguinal lymph nodes or to the accessory axillary lymph nodes. In some cases, it drains to both lymph nodes simultaneously or it may rarely drain only to the medial iliac lymph nodes. The fourth mammary gland with one or two and rarely three lymphatic vessels usually drains to the accessory inguinal lymph nodes. It may rarely drain only to the medial iliac lymph nodes. Mammary lymphatic vessels that cross the midline and lymphatic connection between the mammary glands were not demonstrated. No differences in the mammary lymph drainage pattern between IL and CT-IL were found.     

Immunohistochemical Characterization of Sympathetic Chain Ganglia (SChG) Neurons Supplying the Porcine mammary Gland

The aim of this study was to investigate the chemical coding of mammary gland‐projecting SChG neurons using double‐labelling immunohistochemistry. Earlier observation showed that after injection of the retrograde tracer fast blue (FB) into the second, right thoracic mamma, FB+ mammary gland‐projecting neurons were found in Th1‐3, Th9‐14 and L1‐4 right SChG. The greatest number of FB+ nerve cell bodies was observed in Th10 (approx. 843) and Th11 (approx. 567). Neurons projecting to the last right abdominal mamma were found in L1‐4 SChG. The greatest number of FB+ neurons was observed in L2 (approx. 1200). Immunohistochemistry revealed that the vast majority of FB+ mammary‐projecting neurons contained immunoreactivities to TH (96.97%) and/or DßH (95.92%). Many TH/DßH‐positive neurons stained for SOM (41.5%) or NPY (33.2%), and less numerous nerve cells expressed VIP (16.9%). This observation strongly corresponds to the results of previous studies concerning the immunohistochemical characterization of nerve fibres supplying the porcine mammary gland.     

The fine structure of the Harderian gland in the ostrich (Struthio camelus)

The Harderian gland of the ostrich (Struthio camelus) is a tubuloalveolar gland containing holocrine secreting epithelial cells. The gland epithelium is composed of two different cell types, which can be classified as type I and type II. These cells contain dense secretory vesicles in their cytoplasm and they are connected laterally with desmosomes. At the basal site of these cells, myoepithelial cells are present. Plasma cells are observed in the subepithelial region of the gland. In the interlobular trabeculae, forming the gland stroma, fibroblasts, blood vessels and nerve fibres are included. Another important finding in the ostrich Harderian gland is the presence of homogeneous material.     

Apoptotic Cell Death, bax and bcl-2 Expression During Sheep Mammary Gland Involution

Involution in ovine mammary tissue was studied by light and electron microscopy, and bax and bcl-2 protein distribution was examined by immunohistochemistry from the last day of lactation until the 8th day of drying off. The mammary gland alveoli were examined and the area of glandular epithelium was evaluated morphometrically. Regression of mammary gland epithelium by apoptosis was first identified 2 days after the end of lactation, and increased until day 8. Bax protein was detected throughout this period and was highest on the eighth day. A weak positive reaction for bcl-2 was observed only on days 1 and 8 after cessation of lactation. It is concluded that sheep mammary gland involution involves cell death by apoptosis and that bcl-2 gene family members are involved in the process.     

Distribution of nitric oxide synthase (NOS) immunoreactivity in the chicken stomach

NOS-immunoactive nerve structures were studied in the domestic hen (Gallus domesticus) stomach using single-labelling immunofluorescence. The examined tissues were collected from 10-12 months old laying hens (n=11), Isabrown and Zielononózka races. The distribution and number of the nerve fibres was examined considering differences between the proventriculus and gizzard, and also between particular layers of the stomach wall. Large or moderate numbers of nerve fibres showing NOS-immunoreactivity (NOS-IR) were found in all major layers of both parts of the stomach. In the muscular layer of the gizzard, NOS-positive nerve cell bodies were also observed. Small or moderate numbers of nerve terminals were distributed around blood vessels. NOS-IR nerve fibres associated with blood vessels were slightly more numerous in the gizzard than in the proventriculus. The widespread occurrence of NOS in both parts of the hen stomach can prove its important role in the control of digestive processes. It seems that there is a reasonable necessity of further, more detailed studies on morphological characteristics of the nitrergic nerve structures the bird alimentary tract, their functional significance and plasticity in different physiological and pathological conditions.     

Bovine teat atresia associated with horn fly (Haematobia irritans irritans (L.))-induced dermatitis

Mammary gland lesions characterized primarily as mammary teat atresia were observed in a Limousin beef cattle herd in eastern Texas. Atresia of multiple teats per mammary gland was reported in first-calf heifers at the time of calving. Pathogens were not identified in eight mammary glands collected at slaughter. Histology of affected glands demonstrated superficial and deep perivascular inflammation and fibrosing dermatitis of teat and mammary gland skin that resulted in formation of the atretic lesions of glandular tissue. Institution of a horn fly (Haematobia irritans irritans (L.)) control program using insecticide-impregnated ear tags was associated with elimination of the problem from the herd.