Stimulation of lacrimal secretion by sympathetic nerve impulses in the rabbit

Direct electrical stimulation of the preganglionic nerve trunk of the ipsilateral superior cervical sympathetic ganglion induced fluid secretion from the cannulated main excretory duct of the non-stimulated rabbit lacrimal gland. The optimum strength and frequency of stimulation were 4 volts and 25 Hz. At this stimulus parameter, the rate of secretion was 2.1 +/- 0.2 microliter/10 min. The sympathetic-induced lacrimal secretion was markedly depressed after intravenous administration of hexamethonium (1 mg/kg) and by post-ganglionic neurectomy. The results suggest that, in addition to parasympathetic nerve activity, sympathetic nerve impulses, which pass through the superior cervical sympathetic ganglion, also play a role in initiating fluid secretion in the rabbit lacrimal gland.     

Vasomotor axons of the lacrimal glands of monkeys and the ultrastructural identification of sympathetic terminals

Summary The ultrastructure of the perivascular axon terminals of the lacrimal gland in monkeys is investigated electronmicroscopically. Evidence is presented to show that axon terminals populated with small granular vesicles (300 to 500 Å) are sympathetic. Large granular vesicles (650 to 1,000 Å) are present in both sympathetic and parasympathetic terminals.Lacrimal arterioles have both sympathetic and parasympathetic axon terminals disposed between the adventitia and media, which do not form neuro-effector junctions. Capillaries and venules are sparsely innervated. Both parasympathetic and sympathetic axons are shown to innervate capillaries.Results from degeneration studies show that sympathetic and parasympathetic terminal axons lie within the cytoplasm of single Schwann cells.     

Nerve terminals and epithelial cell variety in the human lacrimal gland

Summary Pieces of tissue taken from three human lacrimal glands were examined electron microscopically. Secretory cells varied according to the electron density and structure of their secretion granules. Secretory cells were arbitrarily categorized as light, medium or dark based on their granule content. Acini were composed of two or all three categories of cells together with myoepithelial cells and lymphocytes.A minority of interstitial nerve fibres contained small granular vesicles characteristic of adrenergic, sympathetic terminals. The appearance of most interstitial and all parenchymal fibres was consistent with that of cholinergic, parasympathetic terminals. Parenchymal terminals were confined to ducts, terminal tubule areas and to serous (dark) cells. A large proportion of parenchymal terminals lay adjacent to myoepithelial cells in the ducts and terminal tubule regions but terminals observed among serous cells were rarely in contact with myoepithelial cells. A possible parasympathetic control of serous secretion, granule production and duct contraction and the autonomy of mucous cells is discussed.The author is indebted to Dr. J. Egeberg of the Anatomy Department B, University of Copenhagen for the provision of embedded material.     

The anatomy and innervation of the mammalian pineal gland

The parenchymal cells of the mammalian pineal gland are the hormone-producing pinealocytes and the interstitial cells. In addition, perivascular phagocytes are present. The phagocytes share antigenic properties with microglial and antigen-presenting cells. In certain species, the pineal gland also contains neurons and/or neuron-like peptidergic cells. The peptidergic cells might influence the pinealocyte by a paracrine secretion of the peptide. Nerve fibers innervating the mammalian pineal gland originate from perikarya located in the sympathetic superior cervical ganglion and the parasympathetic sphenopalatine and otic ganglia. The sympathetic nerve fibers contain norepinephrine and neuropeptide Y as neurotransmitters. The parasympathetic nerve fibers contain vasoactive intestinal peptide and peptide histidine isoleucine. Recently, neurons in the trigeminal ganglion, containing substance P, calcitonin gene-related peptide, and pituitary adenylate cyclase-activating peptide, have been shown to project to the mammalian pineal gland. Finally, nerve fibers originating from perikarya located in the brain containing, for example, GABA, orexin, serotonin, histamine, oxytocin, and vasopressin innervate the pineal gland directly via the pineal stalk. Biochemical studies have demonstrated numerous receptors on the pinealocyte cell membrane, which are able to bind the neurotransmitters located in the pinealopetal nerve fibers. These findings indicate that the mammalian pinealocyte can be influenced by a plethora of neurotransmitters.     

Ultrastructural localization of acetylcholinesterase (AChE) activity in the chicken Harderian gland

Localization of acetylcholinesterase (AChE) was investigated in the chicken Harderian gland at the electron microscopic level. Nerve cells in the pterygopalatine ganglion showed AChE activity. They had a pale and large nucleus which was round or oval in shape. Reaction product of AChE was detected between the nuclear envelopes; in the cisterna of rough endoplasmic reticulum and the lumen of the Golgi lamellae, and on the plasma membrane of the nerve cell. In the interstitium of the gland, nerve fibers showing AChE activity were easily found. They were often seen in the perivascular space and between plasma cells. These nerve fibers had varicosities in contact with plasma cells and the endothelium or the smooth muscle fiber of the blood vessels. AChE-positive varicosities or terminals contained many small clear vesicles (about 50nm in diameter) and a few large dense-cored vesicles (about 100 nm in diameter). No contacts of nerve fibers with acinar cells or the ductal epithelium were observed in the present study. Our data indicate that cholinergic nerves play distinct roles in the regulation of the immune function of the chicken Harderian gland.     

Secretory responses in granular ducts and acini of submandibular glands in vivo to parasympathetic or sympathetic nerve stimulation in rats

Summary The roles of sympathetic and parasympathetic nerves in the secretion of saliva from submandibular glands of rats have been tested by electrical stimulation of either nerve for 1 h unilaterally in separate animals. The flows of saliva thereby induced and their protein content were monitored. Structural changes in each gland were assessed by light- and electron microscopy and compared with the unstimulated contralateral control gland, and the extent of the changes was determined morphometrically. Sympathetic nerve stimulation induced a relatively low flow of saliva that was rich in protein and was accompanied by extensive degranulation from both acinar and granular duct cells. In contrast parasympathetic nerve stimulation induced a considerable flow of saliva that had a low protein content and no detectable degranulation occurred from the secretory cells. It is possible, therefore, that some protein in parasympathetic saliva may have arisen from a non-granular pathway.     

Fluorescence and electron microscopic evidence for the dual innervation of the iris sphincter muscle of the rabbit

Summary The sphincter zone of the rabbit iris sometimes contains terminals with small granular vesicles. These terminals correspond to yellowish-green fluorescent structures in the sphincter zone. The paired arrangement of a terminal containing these vesicles and one full of agranular vesicles might indicate dual innervation of sphincter muscles by sympathetic and parasympathetic fibers. The sympathetic component probably exerts an inhibitory action on the sphincter muscles.This investigation was supported by a research grant from the Ministry of Education, Japan.     

Vasoactive intestinal peptide (VIP)-like immunoreactivity in the human sympathetic ganglia

Vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibres and terminals, neurons and small granule containing cells were observed in human lumbal sympathetic ganglia. Electron-microscopically VIP-IR was localized in the large dense-cored vesicles in nerve terminals and on the membranes of the Golgi complexes in the neurons. A small population of principal ganglion cells was surrounded by VIP-IR nerve terminals. Most of these neurons contained acetylcholinesterase (AChE) enzyme but were not tyrosine hydroxylase-immunoreactive (TH-IR). All VIP-IR ganglion cells and most of the nerve fibres contained AChE but not TH-IR. It appears that in human sympathetic ganglia VIP is localized in the cholinergic neurons and nerve fibres and that the VIP-IR nerve terminals innervate mainly the cholinergic subpopulation of the sympathetic neurons.     

Vasoactive intestinal peptide (VIP)-like immunoreactivity in the human sympathetic ganglia

Summary Vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibres and terminals, neurons and small granule containing cells were observed in human lumbal sympathetic ganglia. Electron-microscopically VIP-IR was localized in the large dense-cored vesicles in nerve terminals and on the membranes of the Golgi complexes in the neurons. A small population of principal ganglion cells was surrounded by VIP-IR nerve terminals. Most of these neurons contained acetycholinesterase (AChE) enzyme but were not tyrosine hydroxylase-immnoreactive (TH-IR). All VIP-IR ganglion cells and most of the nerve fibres contained AChE but not TH-IR. It appears that in human sympathetic ganglia VIP is localized in the cholingergic neurons and nerve fibres and that the VIP-IR nerve terminals innervate mainly the cholinergic subpopulation of the sympathetic neurons.     

Adrenergic control of lacrimal secretion in rabbits

Stimulation of the preganglionic trunk of the ipsilateral superior cervical sympathetic ganglion with square wave electrical pulses produced secretion from the main excretory duct of the rabbit lacrimal gland. The secretion was inhibited by propranolol (1 mg/kg) and metoprolol (10 mg/kg) but not by phenoxybenzamine (2 mg/kg) and atropine (25 micrograms/kg). The results indicate that the sympathetically induced secretion in the rabbit lacrimal gland occurs through an adrenergic mechanism. The adrenergic receptors in the lacrimal gland is most likely of beta 1-type.     

Transient involvement of enkephalins in both the sympathetic and parasympathetic innervations of the submandibular gland of rats

Summary A time course study with enkephalin(Enk)-like immunoreactivity has revealed that nerve fibers intensely immunoreactive for Enk-8 appeared transiently only during the postnatal week 2 and 4 within the acini as well as in the inter- and intralobular connective tissues of the submandibular gland of rats. At these stages numerous nerve fibers immunoreactive for tyrosine hydroxylase (TH) appeared also in the inter- and intralobular connective tissues and within the acini. Coincidently with these postnatal stages, abundant Enk-immunoreactive principal ganglion cells appeared in the superior cervical ganglion. These were not immunoreactive for neuropeptide tyrosine (NPY). A substantial number of Enk-immunoreactive ganglion cells were also present in the submandibular ganglia at these younger postnatal stages. Superior cervical ganglionectomy at these stages resulted in a marked decrease in number of the inter- and intralobular Enk-immunoreactive nerve fibers, a slight decrease in number of the intraacinar Enk-immunoreactive nerve fibers, and almost complete disappearance of intraglandular TH-immunoreactive nerve fibers. Immuno-electron-microscopic analysis revealed that Enk-immunoreactive nerve fibers in the submandibular gland were identified as electron-dense neuronal profiles enclosed by Schwann cells in the inter- and intralobular connective tissues and those directly apposed to secretory cells within the acini. They contained small clear vesicles mixed with some large granular vesicles. After postnatal week 6, no Enk-immunoreactive nerve fibers were detected in the submandibular gland, and no TH-immunoreactive nerve fibers were seen within the acini, while TH-immunoreactive nerve fibers remained numerous in the inter- and intralobular connective tissues. These findings indicate that both sympathetic and parasympathetic nerve fibers exhibit Enk-like immunoreactivity transiently during postnatal weeks 2 and 4. It is further indicated that the inter- and intralobular nerve fibers lose Enk-like immunoreactivity while the intraacinar fibers disappear at the adult stage.     

Localization of neuropeptide Y in human sympathetic ganglia: Correlation with met-enkephalin, tyrosine hydroxylase and acetylcholinesterase

Summary The relationships of immunoreactive neuropeptide Y, enkephalin and tyrosine hydroxylase, on the one hand, and acetylcholinesterase histochemical activity, on the other, were studied in human lumbar sympathetic ganglia. Two thirds of the ganglion cells contained immunoreactive neuropeptide Y. Electron microscopically the immunoreaction was localized in the Golgi apparatus and in large dense-cored vesicles in the nerve endings. Most of the neuropeptide-containing neurons and nerve fibres were also reactive for tyrosine hydroxylase. Nerve fibres reactive for neuropeptide Y were found around ganglion cells regardless of their transmitter contents, whereas enkephalin-reactive nerve terminals surrounded only acetylcholinesterase-containing neurons. The results demonstrate that neuropeptide Y is colocalized with noradrenaline in most of the human sympathetic neurons and that the nerve fibres may innervate selectively the noradrenergic and cholinergic subpopulations of ganglion cells depending on the transmitters of the nerves.     

Catecholaminergic innervation of the rat adrenal cortex

Summary The zona glomerulosa of the rat adrenal gland is innervated by catecholaminergic nerves. Using histofluorescence techniques, we observed catecholaminergic plexuses surrounding adrenal capsular and subcapsular blood vessels. Individual varicose nerve fibers that branched off these plexuses were distributed among adrenal glomerulosa cells. This innervation was permanently eliminated after neonatal sympathectomy with guanethidine or 6-hydroxydopamine, but was not affected by ligation of the splanchnic nerve or extirpation of the suprarenal ganglion. At the ultrastructural level, axonal varicosities were commonly observed in close proximity to glomerulosa cells and blood vessels. Nerve fibers and varicosities were found to contain small (30–60 nm) clear vesicles as well as large (60–110 nm) and small (30–60 nm) dense-cored vesicles. In tissue fixed for the dichromate reaction with or without pretreatment with the false transmitter 5-hydroxydopamine, many nerve terminals contained numerous small dense-cored vesicles which are thought to contain catecholamines. These results establish the anatomical substrate for the catecholaminergic innervation of the rat adrenal cortex.     

Light and electron microscopic histochemistry of the monoamines in the human foetal sympathetic ganglion in culture

Synopsis Sympathetic ganglia of 13 to 19-week-old human foetuses were cultured in small pieces with and without nerve growth factor for up to 5 weeksin vitro. The cultures were studied using phase-contrast, fluorescence and electron microscopy. Monoamines were demonstrated with the formaldehyde-induced fluorescence method, with and without pretreatment of the cultures with catecholamines or monoamine oxidase inhibitor.In the long-term cultures, primitive sympathetic cells, sympathicoblasts of types I and II, and young sympathetic neurons showed a fine structure identical to that described earlierin vivo. There were virtually no satellite or Schwann cells in the cultures. The neurons showed a considerable capacity to grow new nerve fibres in culture, even without nerve growth factor. Nerve terminals with accumulations of other nervous structures. Large granular vesicles were regularly found in the sympathicoblasts after glutaraldehyde-osmium tetroxide fixation. After permanganate fixation, dense-cored vesicles typical of adrenergic neurons were not seen, either in the perikarya, or in the processes, although it was possible to demonstrate specific fluorescence. No small intensely fluorescent (SIF) cells were observed.Variable formaldehyde-induced fluorescence was observed in the nerve cell perikarya and nerve fibres. The intensity of the fluorescence increased after treatment of the cultures with monoamine oxidase inhibitor and after incubation with catecholamines.     

The fine structure of the ganglia of the guinea-pig trachea

Summary The parasympathetic ganglia of the guinea-pig trachea have been investigated by scanning and transmission electron microscopy. They are covered by a continuous perineurium and connective tissue is found between the neural elements. Blood vessels inside the ganglia have continuous endothelia and are sometimes accompanied by pericytes and a sheath of perineurial cells. Individual neuronal cell bodies and large processes are almost completely covered by a thin layer of satellite cells, except for very small areas that directly face the basal lamina and connective tissue space. Nerve fibres are also completely and individually ensheathed by Schwann cell processes; naked fibres are not found. In some regions of the nerve cell body, there are complex interdigitations between short neuronal processes and satellite cells. Large differences in the size of neurons may indicate the presence of different neuronal populations. Nerve endings containing mainly small clear vesicles are the most common type, and these form synapses on dendrites, but some profiles have many large granular vesicles. These ganglia resemble other parasympathetic, sympathetic and sensory ganglia and not the enteric ganglia. However, an unusual feature of the cytoplasm of the satellite and Schwann cells is the abundance of 10 nm intermediate filaments.     

Stimulation-depletion of serotonin and noradrenaline from vesicles of sympathetic nerves in the pineal gland of the rat

Summary The pineal gland of the rat receives a rich nervous supply originating from the superior cervical ganglia. These fibers contain serotonin in addition to their neurotransmitter, noradrenaline. Cytochemical studies at the ultrastructural level have shown that both amines are present in the cores of the granular vesicles that are characteristic of these nerves. It is presently shown that the bilateral electrical stimulation of the preganglionic fibers innervating the ganglia markedly reduces the number of small sites reacting cytochemically for both noradrenaline and serotonin, these sites corresponding to the cores of small granular vesicles, while the larger reactive sites (cores of large vesicles) remain unaltered. The vesicles are retained in nerve terminals after stimulation, as observed in conventionally processed tissues, although with altered sizes and shapes. Apart from these cytochemical and structural changes, nerve stimulation also reduces the endogenous noradrenaline content of the pineal gland. Thus, both noradrenaline and serotonin are released from their storage sites in pineal sympathetic nerves after electrical stimulation in vivo. This suggests the possibility that several substances with presumed transmitter or modulatory functions might be simultaneously released by nerve impulses from a given nerve terminal.     

Sympathetic activation increases basilar arterial blood flow in normotensive but not hypertensive rats

The close apposition between sympathetic and parasympathetic nerve terminals in the adventitia of cerebral arteries provides morphological evidence that sympathetic nerve activation causes parasympathetic nitrergic vasodilation via a sympathetic-parasympathetic interaction mechanism. The decreased parasympathetic nerve terminals in basilar arteries (BA) of spontaneously hypertensive rat (SHR) and renovascular hypertensive rats (RHR) compared with Wistar-Kyoto rats (WKY), therefore, would diminish this axo-axonal interaction-mediated neurogenic vasodilation in hypertension. Increased basilar arterial blood flow (BABF) via axo-axonal interaction during sympathetic activation was, therefore, examined in anesthetized rats by laser-Doppler flowmetry. Electrical stimulation (ES) of sympathetic nerves originating in superior cervical ganglion (SCG) and topical nicotine (10-30 μM) onto BA of WKY significantly increased BABF. Both increases were inhibited by tetrodotoxin, 7-nitroindazole (neuronal nitric oxide synthase inhibitor), and ICI-118,551 (β(2)-adrenoceptor antagonist), but not by atenolol (β(1)-adrenoceptor antagonist). Topical norepinephrine onto BA also increased BABF, which was abolished by atenolol combined with 7-nitroindazole or ICI-118,551. Similar results were found in prehypertensive SHR. However, in adult SHR and RHR, ES of sympathetic nerves or topical nicotine caused minimum or no increase of BABF. It is concluded that excitation of sympathetic nerves to BA in WKY causes parasympathetic nitrergic vasodilation with increased BABF. This finding indicates an endowed functional neurogenic mechanism for increasing the BABF or brain stem blood flow in coping with increased local sympathetic activities in acutely stressful situations such as the "fight-or-flight response." This increased blood flow in defensive mechanism diminishes in genetic and nongenetic hypertensive rats due most likely to decreased parasympathetic nitrergic nerve terminals.     

An immuno-electron-microscopic study of the localization of VIP-like immunoreactivity in the adrenal gland of the rat

Summary VIP-like immunoreactivity was revealed in a few chromaffin cells, medullary ganglion cells and a plexus of varicose nerve fibers in the superficial cortex and single varicose fibers in the juxtamedullary cortex and the medulla of the rat adrenal gland. VIP-like immunoreactive chromaffin cells were polygonal in shape without any distinct cytoplasmic processes and they appeared solitarily. Their cytoplasm contained abundant granular vesicles having a round core and the immunoreactive material was localized to the granular core. VIP-immunoreactive ganglion cells were multipolar and had large intracytoplasmic vacuoles. The immunoreactive material was localized not only in a few granular vesicles but also diffusely throughout the axoplasm. VIP-immunoreactive varicose nerve fibers in the superficial cortex were characterized by abundant small clear vesicles and some large granular vesicles, while those in the juxtamedullary cortex and medulla and the ganglionic processes were characterized by abundant large clear vesicles, as well as the same vesicular elements as contained in the nerves in the superficial cortex. The immunoreactive material was localized on the granular cores and diffusely in the axoplasm in both nerves. Based on the similarity and difference in the composition of the vesicles contained in individual nerves, it is likely that the VIP-immunoreactive nerve fibers in the medulla and the juxtamedullary cortex are derived from the medullary VIP-ganglion cells, while those in the superficial cortex are of extrinsic origin. The immunoreactive nerve fibers in both the cortex and the medulla were often in direct contact with cortical cells and chromaffin cells, where no membrane specializations were formed. The immunoreactive nerve fibers were sometimes associated with the smooth muscle cells and pericytes of small blood vessels in the superficial cortex. In addition they were often seen in close apposition to the fenestrated endothelial cells in the cortex and the medulla, only a common basal lamina intervening. Several possible mechanisms by which VIP may exert its effect in the adrenal gland are discussed.     

Amine-containing peptidergic synapses in a parasympathetic ganglion?

Summary Following fixation with the modified chromaffin reaction of Tranzer and Richards (1976) unusual nerve profiles were found forming synapses on neurons in a parasympathetic ganglion intrinsic to the rabbit rectococcygeus muscle. Synaptic profiles were remarkable in having very high proportions of large filled vesicles. In addition to numerous large filled vesicles, many of these profiles contained a few chromaffin-reactive small granular vesicles. It is suggested that these profiles contain and may release both peptides and amines or amine-like substances which could mediate or modulate synaptic transmission in the 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.