Ultrastructural observations on the central innervation of the guinea-pig pineal gland

Summary In the present study the central innervation of the guinea-pig pineal gland was investigated. The habenulae and the pineal stalk contain myelinated and non-myelinated nerve fibres with few dense-cored and electron-lucent vesicles. Some myelinated fibres leave the main nerve fibre bundles, lose their myelin-sheaths and terminate in the pineal gland. Although direct proof is lacking, the non-myelinated fibres appear to end near the site where the bulk of the myelinated fibres are located. Here a neuropil area exists where synapses between non-myelinated fibre elements are abundant. Neurosecretory fibres were also seen. The results support the concept of functional interrelationships between hypothalamus, epithalamus and the pineal gland.     

Activity pattern of suboesophageal ganglion cells innervating the salivary glands of the locust Locusta migratoria

The salivary gland of the locust, Locusta migratoria, is innervated from the suboesophageal ganglion by two neurones, SN1 and SN2 which innervate the gland via the salivary gland nerve (nerve 7B of the suboesophageal ganglion). In addition, like most other peripheral nerves of the head, this nerve carries on its outer surface axons and neurohaemal terminal ramifications of the so called satellite nervous system, established by a group of neurosecretory cells also located in the suboesophageal ganglion. These superficial collaterals ramify over the nerve from its origin in the head to its terminals within the gland in the thoracic segments.Nerve 7B was recorded chronically in freely moving locusts. Both salivary neurones are active during and shortly before feeding, as defined by continuous rhythmic activity of the mandibular closer muscle (M9). The activity of the salivary neurones, particularly that of SN2, thus resembles that of the satellite neurones as described recently. While SN2 ceases firing at the end of a feeding bout, SN1 continues firing for a short period. Also, SN1 fires short bursts of impulses for a few minutes following the end of a feeding bout. Similar bursts also occur at random intervals during the long-lasting phases between feeding events.Abbreviations SN1 salivary neurone 1 - SN2 salivary neurone 2 - M9 mandibular closer muscle - DUM dorsal unpaired median - LMN labral median nerve     

Innervation of the rat pineal gland by pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres

Pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres were demonstrated in the rat pineal gland. These fibres entered the pineal gland through the conarian nerve at the distal tip of the gland. A high density of the fibres was observed in the capsule of the gland, from where the immunoreactive elements penetrated into the pineal perivascular spaces and parenchyma. The majority of PACAP-immunoreactive nerve fibres also contained calcitonin gene-related peptide (CGRP). Some PACAP-immunoreactive nerve fibres contained neuropeptide Y (NPY), but only occasionally was PACAP colocalized with vasoactive intestinal peptide (VIP). After removal of both superior cervical ganglia, a high number of PACAP-containing nerve fibres were still present in the gland. In the nervous system PACAP is present in two isoforms, PACAP-38 and PACAP-27. The concentration of PACAP-38 in the superficial pineal gland was determined by radioimmunoassay to be 20.4 pmol/g tissue at midday and 18.9 pmol/g tissue at midnight. The concentration of PACAP-27 was only about 3% of the concentration of PACAP-38. In summary, this study is the first demonstration of a PACAP-containing innervation of the rat pineal gland. The PACAP concentration in the pineal gland does not exhibit a day-night difference. The colocalization of PACAP with calcitonin gene-related peptide in the pincalopetal nerve fibres indicates that the majority of PACAP-immunoreactive nerve fibres might originate from the trigeminal ganglion.     

Effects of heterologous cross-suture between the postganglionic sympathetic and the preganglionic parasympathetic nerve trunks of submandibular glands in cats

Summary After sectioning the postganglionic adrenergic sympathetic nerve trunk for the submandibular gland, as close to the submandibular artery as practicable, its central end was sutured to the peripheral end of the preganglionic cholinergic parasympathetic nerve trunk for the gland, the chorda, which had been sectioned where it left the lingual nerve. The effects of this heterologous cross-sature were studied at different times, up to 1 year afterwards, by assessing the physiological and pharmacological responses of the glands and the neuro-histochemical changes in the nerve trunks and in the nerves within the glands.In all cases adrenergic sympathetic nerves grew across the site of suture and down the erstwhile cholinergic parasympathetic trunk, eventually to develop connections in the gland. In some cases the functional adrenergic reinnervation of the submandibular gland appeared to result exclusively or predominantly from the direct downgrowth of adrenergic axons to the gland, via the crossed nerves. In other cases however, in addition to a direct glandular reinnervation, there was some physiological and morphological evidence which suggested that possible heterogenous synaptic contacts may have been created between postganglionic sympathetic axons and cholinergic ganglion cells in the chorda nerve.This work was supported by a grant from the Joint Research Committee, King's College Hospital.     

Efferent projections from the lateral geniculate nucleus to the pineal complex of the Mongolian gerbil (Meriones unguiculatus)

Summary The intergeniculate leaflet of the lateral geniculate nucleus is considered to modulate circadian activity rhythms probably mediated by a direct neuronal connection to the suprachiasmatic nucleus. The present study in the gerbil demonstrates, by anterograde tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L), the existence of an additional neuronal projection from a subportion of the lateral geniculate nucleus, involving the intergeniculate leaflet, directly to the pineal gland. PHA-L-immunoreactive nerve fibers originating from perikarya at the injection site were located under the optic tract projecting towards the midsagittal plane. Delicate PHA-L-immunoreactive nerve fibers were observed in the posterior paraventricular thalamic nucleus, precommissural nucleus, olivary pretectal nucleus, anterior and posterior pretectal nuclei, and posterior commissure. Single fibers could be followed from the caudal part of the medial habenular nucleus and the pretectal area into the rostral part of the deep pineal gland. Other fibers continued through the posterior commissure into the contralateral hemisphere to terminate in the same structures as on the ipsilateral side. From the posterior commissure, small bundles of thick fibers entered the deep pineal gland where they arborized among the endocrine cells. A few nerve fibers were observed in the habenular commissure and the pineal stalk, but no fibers were identified in the superficial pineal. This direct geniculo-pineal connection suggests that the pineal gland is directly influenced by the optic system.     

Immunohistochemical localization of substance P in the pineal gland of the domestic pig

An immunohistochemical study of the pig pineal gland was carried out using polyclonal rabbit antiserum raised against substance P (SP). The pineal glands were taken from the newborn, 21-day- and 7-month-old female pigs. Immunoreactive nerve fibers were observed in the pineal gland as well as in the posterior commissure and habenular areas. The bundles of SP-immunoreactive fibers were also seen in the subependymal layer of the pineal tissue. The single SP-immunoreactive nerve fibers and few small bundles of nerve fibers were located with equal density throughout the pineal gland, in the connective tissue septa and in the parenchyma. SP-immunoreactive cell bodies were observed in the medial habenular nucleus. The obtained results point to this nucleus as one of the central sources of SP innervation in the pig pineal gland. The study did not show any differences in the distribution and the density of SP-immunoreactive nerve fibers between newborn, 21-day- and 7 month-old pigs.     

Ultrastructure and maturation of a sex pheromone gland in the female German cockroach, Blattella germanica

A volatile sex pheromone is produced in an adult female-specific gland located on the anterior of the last abdominal tergite of the female German cockroach, Blattella germanica (L.). In this area, the cuticle forms deep depressions in which a large number of cuticular orifices are located. The cuticular orifices are connected to secretory cells via cuticular ducts surrounded by duct cells. The pheromone gland exhibits a clear developmental maturation in relation to sexual maturation of the female. The secretory cells of a newly formed gland in the imaginal female are small and contain few secretory vesicles. The amount of extractable pheromone in the gland is low on day-0 but it increases with age and peaks on day-6. The secretory cells in a mature day-6 gland are characterized by a large number of electron-lucid secretory vesicles. abundant RER and SER, a large nucleus and a long, convoluted end apparatus which is lined with numerous microvilli. The contents of the secretory vesicles are exocytosed into extracellular reservoirs at the base of microvilli and then transported to the cuticular surface through the long ducts. The supportive function of the duct cell in the glandular organization and developmental regulation of the gland are discussed.     

Localization of monoamine nerve fibres by formaldehyde fluorescence histochemistry in the posterior salivary duct and gland of Octopus vulgaris.

A bright yellow-green specific fluorescence is induced by formaldehyde histochemistry for monoamines in the secretory nerve trunks of the Octopus vulgaris posterior salivary duct, and in their ramification in the gland tubules. In contrast, the motor nerve trunks of the duct contain few fluorescent elements. The muscular and connective coat of the duct is provided with fluorescent globular and varicose structures, of various sizes and colours, which become numerous in the duct branches. At least some of these peripheral structures belong to varicose monoamine nerve fibres. In the gland, on the contrary, the muscle cells surrounding the tubules are not supplied with fluorescent nerve fibres.     

The safe face lift with bony anatomic landmarks to elevate the SMAS

The risk for facial nerve injury has been reported to be increased with the inclusion of superficial musculoaponeurotic system (SMAS) elevation as compared with a skin-only face lift. The facial nerve courses through the parotid gland. The SMAS is elevated superficial to the parotid gland. However, in elevating the SMAS anterior to the parotid gland, the facial nerve is at risk of injury where its branches emerge from the anterior edge of the parotid gland. The purpose of this study was to identify bony anatomic landmarks to predict the location of the anterior edge of the parotid gland to avoid injury to the facial nerve branches as they exit the parotid gland. The authors dissected 20 cadaver face halves to determine bony landmarks-the masseteric tuberosity and the inferior lateral orbital rim-to predict the location of the anterior parotid edge. Then they measured the anterior edge of the parotid gland in relation to the vector formed between these two bony landmarks. They identified and measured the most anterior portion of the parotid gland in relation to this vector. Then the most posterior aspect of the parotid gland in relation to this vector was measured. In the 20 dissections, the authors found the most anterior portion of the parotid gland to be 2.7 +/- 1.0 mm anterior to the vector from the inferior lateral orbital rim to the masseteric tuberosity. The most posterior part of the anterior edge of the parotid gland in relation to this vector was found to be 1.0 +/- 1.5 mm posterior to this vector. The parotid gland measured an average of 38.8 +/- 3.5 mm in width from the tragus to the anterior parotid edge. In elevating the SMAS with a face lift, the facial nerve branches can be predicted to exit the anterior edge of the parotid gland, which can be located 38.8 mm anterior to the tragus and near the vector from the inferior lateral orbital wall to the masseteric tuberosity.     

Innervation of the C cells of chicken ultimobranchial glands studied by immunohistochemistry, fluorescence microscopy, and electron microscopy

Innervation of the ultimobranchial glands in the chicken was investigated by immunohistochemistry, fluorescence microscopy and electron microscopy. The nerve fibers distributed in ultimobranchial glands were clearly visualized by immunoperoxidase staining with antiserum to neurofilament triplet proteins (200K-, 150K- and 68K-dalton) extracted from chicken peripheral nerves. The ultimobranchial glands received numerous nerve fibers originating from both the recurrent laryngeal nerves and direct vagal branches. The left and right sides of the ultimobranchial region were asymmetrical. The left ultimobranchial gland had intimate contact with the vagus nerve trunk, especially with the distal vagal ganglion, but was somewhat separated from the recurrent nerve. The right gland touched the recurrent nerve, the medial edge being frequently penetrated by the nerve, but the gland was separated from the vagal trunk. The left gland was innervated mainly by the branches from the distal vagal ganglion, whereas the right gland received mostly the branches from the recurrent nerve. The carotid body was located cranially near to the ultimobranchial gland. Large nerve bundles in the ultimobranchial gland ran toward and entered into the carotid body. By fluorescence microscopy, nerve fibers in ultimobranchial glands were observed associated with blood vessels. Only a few fluorescent nerve fibers were present in close proximity to C cell groups; the C cells of ultimobranchial glands may receive very few adrenergic sympathetic fibers. By electron microscopy, numerous axons ensheathed with Schwann cell cytoplasm were in close contact with the surfaces of C cells. In addition, naked axons regarded as axon terminals or "en passant" synapses came into direct contact with C cells. The morphology of these axon terminals and synaptic endings suggest that ultimobranchial C cells of chickens are supplied mainly with cholinergic efferent type fibers. In the region where large nerve bundles and complex ramifications of nerve fibers were present, Schwann cell perikarya investing the axons were closely juxtaposed with C cells; long cytoplasmic processes of Schwann cells encompassed large portions of the cell surface. All of these features suggest that C-cell activity, i.e., secretion of hormones and catecholamines, may be regulated by nerve stimuli.     

Localisation of monoamines in nerves of the posterior salivary gland and salivary centre in the brain of Octopus

Summary Slices of the posterior salivary gland and of the superior buccal lobe of the brain of Octopus vulgaris take up ³H-5-hydroxytryptamine in vitro. By light and electron microscopical radioautography the uptake is localised in certain neuronal perikarya and axonal varicosities in the superior buccal lobe, and in nerves that end in the secretory tubules of the posterior salivary gland. These structures do not incorporate ³H-noradrenaline. After formaldehyde histochemistry, monoamine fluorescence is found in some neuronal perikarya of the superior buccal lobe, and in nerves entering the secretory tubules of the gland. The posterior salivary gland nerve, which originates in the superior buccal lobe and supplies the gland, shows a pronounced accumulation of fluorescence on the proximal side of a ligature applied in vivo. It is suggested that monoamines are transported from the brain to the gland by the posterior salivary gland nerves.J. B. would like to thank the Science Research Council, Great Britain, for financial support.     

Gamma-aminobutyric acid (GABA) immunoreactivity in the mouse adrenal gland

Gamma-aminobutyric acid (GABA) immunoreactivity was revealed by immunocytochemistry in the mouse adrenal gland at the light and electron microscopic levels. Groups of weakly or faintly GABA immunoreactive chromaffin cells were often seen in the adrenal medulla. By means of immunohistochemistry combined with fluorescent microscopy, these GABA immunoreactive chromaffin cells showed noradrenaline fluorescence. The immunoreaction product was seen mainly in the granular cores of these noradrenaline cells. These results suggest the co-existence of GABA and noradrenaline within the chromaffin granules. Sometimes thick or thin bundles of GABA immunoreactive nerve fibers with or without varicosities were found running through the cortex directly into the medulla. In the medulla, GABA immunoreactive varicose nerve fibers were numerous and were often in close contact with small adrenaline cells and large ganglion cells; a few, however, surrounded clusters of the noradrenaline cells, where membrane specializations were formed. Single GABA immunoreactive nerve fibers, and thin or thick bundles of the immunoreactive varicose nerve fibers ran along the blood vessels in the medulla. The immunoreaction deposits were observed diffusely in the axoplasm and in small agranular vesicles of the GABA immunoreactive nerve fibers. Since no ganglion cells with GABA immunoreactivity were found in the adrenal gland, the GABA immunoreactive nerve fibers are regarded as extrinsic in origin.     

Structure and Role of the Renette Cell and Caudal Glands in the Nematode Sphaerolaimus gracilis (Monhysterida)

Ultrastructure of the renette cell and caudal glands was studied in the free-living aquatic nematode Sphaerolaimus gracilis. The renette cell occurred posterior to the esophageal-intestinal junction and opened through an ampulla to a ventral pore behind the nerve ring. The caudal gland system of the tail consisted of two gland cells opening through separate pores and 2 to 3 other gland cells of a different type opening through a common pore. The renette cell and the two caudal gland cells were similar and both contained secretory granules, 0.5-1.5 μm in diameter. The material released attached the nematode to the substrate. The renette ampulla was surrounded by a specialized cell, the ampulla cell, which had characteristics of myoepithelium. A plug or valve structure connected to the ampulla cell may regulate the output of the secretory material. The ampulla cell is able to contract and thus is probably under direct neuronal control. Other cells in the renette ampulla region of body cavity were termed supporting cells. Living, cold-relaxed nematodes were attached to sediment particles in the renette pore region and at the tail tip. Release from sediment particles was mechanical at the renette cell discharge site but appeared to be chemical at the caudal gland. In behavioral experiments, nematodes in a water current had the ability to release a thread from the caudal glands while maintaining contact with a sediment particle attached to the tail end. If the thread was strong enough, it also could be used to change location. Nematodes anchored by the thread from the caudal glands to a sediment particle could float in water currents until they attached themselves to another sediment particle with the help of secretions from the renette cells.     

Innervation of the cat pineal gland by neuropeptide Y-immunoreactive nerve fibers: an experimental immunohistochemical study

An immunohistochemical study of the cat pineal gland was performed using a rabbit polyclonal antibody directed against neuropeptide Y (NPY) and an antibody directed against the C-terminal flanking peptide of neuropeptide Y (CPON). Numerous NPY- and CPON-immunoreactive (IR) nerve fibers were demonstrated throughout the gland and in the pineal capsule. The number of IR nerve fibers in the capsule was high and from this location fibers were observed to penetrate into the gland proper via the pineal connective tissue septa, often following the blood vessels. From the connective tissue septa IR fibers intruded into the parenchyma between the pinealocytes. Many IR nerve fibers were observed in the pineal stalk and in the habenular as well as the posterior commissural areas. The number of NPY/CPON-IR nerve fibers in pineal glands from animals bilaterally ganglionectomized two weeks before sacrifice was low. The source of most of the extrasympathetic NPY/CPONergic nerve fibers is probably the brain from where they enter the pineal via the pineal stalk. However, an origin of some of the fibers from parasympathetic ganglia cannot be excluded due to the presence of a few IR fibers in the pineal capsule of ganglionectomized animals. It is concluded that the cat pineal is richly innervated with NPYergic nerve fibers mostly of sympathetic origin. The posttranslational processing of the NPY promolecule results in the presence of both NPY and CPON in intrapineal nerve fibers.     

Effects of surgical denervations on the autonomic nerves in parotid glands of dogs

Summary The innervation of the dog's parotid has been studied by cholinesterase staining and catecholamine fluorescence. In normal glands cholinergic and adrenergic nerves are plentiful around acini, muscular blood vessels, and to a lesser extent striated ducts. The main ducts, although surrounded by many cholinesterase-positive nerves, are associated with few adrenergic nerves. Severance of the classical parasympathetic post-ganglionic nerve to the gland, the auriculo-temporal, caused a moderate loss of cholinesterase-positive nerves. When this procedure was combined with section of the nerves on the internal maxillary artery there was a greater loss. Fewest cholinesterase-positive nerves remained when, in addition to these two procedures, the facial nerve was cut. These findings support the concept that all three sets of nerves contain some post-ganglionic parasympathetic fibres for the dog's parotid. The source of the remaining nerves is unknown. Preganglionic parasympathetic denervation by section of the tympanic branch of the glossopharyngeal nerve did not reduce the number of cholinesterase-positive nerves. None of these parasympathetic denervations caused reduction of adrenergic nerves, indicating that they do not travel to the gland with the parasympathetic nerves. After superior cervical ganglionectomy a few scattered fluorescent nerves remained in the gland; their origin is unknown.     

Elongation of mouse spinal cord axons in isogenic grafts of the sciatic nerve, skeletal muscle and submaxillary gland

Isografts of sciatic nerve, skeletal muscle, submaxillary gland and, as control experiments, of optice nerve, were transplanted into the non transected spinal cord of young albino mice, through a punctiform pial aperture. Under these conditions, local cellular reactions were reduced and the sensori motor behavior of the operated animals remained apparently undisturbed throughout the experimental period. Within a few days, axonal sprouts issuing mainly from the terminal clubs of intraspinal nerve fibres severed by the grafting procedure were seen elongating and growing into--and presumably throughout--the nervous as well as the muscular and glandular transplants. The Schwann cells of these grafts, either sedentary or migrating towards the cord and intermingling with host reactive glial cells, appeared to guide the growth of the axonal sprouts they ensheathed (from day 3 to day 10) and generally myelinated (as early as day 6). Optic nerve transplants, lacking Schwann cells, were never reinnervated. Furthermore, in control microinjuries without grafting, limited growth of axonal sprouts was observed only when a few host Schwann cells were present. Mouse spinal neurons, therefore, demonstrate a marked capacity for regrowth when minimal damage to the spinal cord is associated with an adequate supply of Schwann cells. In contrast, host as well as transplanted glial cells, were unable, at least when they were not associated with Schwannian elements, to promote regenerative expression of these central neurons.     

Ultrastructure of the rat pineal stalk

The ultrastructure of the rat pineal stalk was described. The pineal stalk contained few pinealocytes, glial cells and numerous nerve fibers. The last were mostly non-myelinated axons, although a few myelinated ones were also observed. Glial cells showed many filaments, mostly in the processes which presented a longitudinal orientation. Other more lamellar processes were found enclosing the axons. The pineal stalk became wider as it reached the body of the gland. Ultrastructurally, this wide region resembled more the pineal body. Bundles of non-myelinated nerve fibers were seen around the pineal stalk.     

Observations on the fine structure,enzyme histochemistry,and innervation of parathyroid gland and ultimobranchial body of Chthonerpeton indistinctum (Gymnophiona,Amphibia)

Summary Fine structural and enzyme histochemical observations on ultimobranchial body and parathyroid gland of the caecilian Chthonerpeton are presented. The cell clusters and follicles of the ultimobranchial body consist mainly of granulated cells which are termed C-cells and obviously belong to the APUD cell series. In the larger follicles additional possibly exhausted degranulated cells and replacement cells occur. A rich supply of nerve fibres has been found in this gland. Frequently nerve terminals were observed to come into synaptic contact with the C-cells. Two categories of nerve fibres occur: a) fibres containing large polymorphic electron dense granules (probably purinergic fibres), b) fibres containing small electron transparent vesicles and a few electron dense granules (probably cholinergic fibres). The parathyroid gland consists of elongated cells (one cell type) poor in organelles and often containing fields of glycogen and lipid droplets. The cells are further characterized by fair amounts of lysosomal enzymes; they are interconnected by maculae adhaerentes and occludentes. No nerves and blood vessels have been found in the parathyroid gland of Chthonerpeton. This study has been supported by the Deutsche Forschungsgemeinschaft We 380/5.     

Autonomic receptors and cyclic nucleotides of rat parotid gland following simultaneous electrical stimulation of its parasympathetic and sympathetic nerves

[3H]dihydroalprenolol and [3H]quinuclidinylbenzilate binding of membranes of rat parotid gland were generally unchanged after 10, 15, 30, or 60 min of simultaneous electrical stimulation of the parasympathetic and sympathetic nerves to the gland, although stimulation of either nerve separately caused nerve-specific changes in both. Concentrations of cyclic nucleotides of the gland were, however, increased significantly from levels of the unstimulated parotid gland. Cyclic GMP showed a 10-fold increase after 10 min of stimulation, whereas only a 2-fold increase in cyclic AMP was found at this time. The increases were maintained, albeit at reduced levels, at 15 and 30 min also but by 60 min both were not different from levels of the unstimulated gland. The increases induced by separate stimulation of each nerve were greater but nerve specific, and the changes induced with simultaneous stimulation tended to reflect a reigning influence of one nerve on the other.