Elevated expression of neuropeptide signaling genes in the eyestalk ganglia and Y-organ of Gecarcinus lateralis individuals that are refractory to molt induction.

Molting is induced in decapod crustaceans via multiple leg autotomy (MLA) or eyestalk ablation (ESA). MLA removes five or more walking legs, which are regenerated and become functional appendages at ecdysis. ESA eliminates the primary source of molt-inhibiting hormone (MIH) and crustacean hyperglycemic hormone (CHH), which suppress the production of molting hormones (ecdysteroids) from the molting gland or Y-organ (YO). Both MLA and ESA are effective methods for molt induction in Gecarcinus lateralis. However, some G. lateralis individuals are refractory to MLA, as they fail to complete ecdysis by 12weeks post-MLA; these animals are in the "blocked" condition. Quantitative polymerase chain reaction was used to quantify mRNA levels of neuropeptide and mechanistic target of rapamycin (mTOR) signaling genes in YO, eyestalk ganglia (ESG), thoracic ganglion (TG), and brain of intact and blocked animals. Six of the seven neuropeptide signaling genes, three of four mTOR signaling genes, and Gl-elongation factor 2 (EF2) mRNA levels were significantly higher in the ESG of blocked animals. Gl-MIH and Gl-CHH mRNA levels were higher in the TG and brain of blocked animals and levels increased in both control and blocked animals in response to ESA. By contrast, mRNA levels of Gl-EF2 and five of the 10 MIH signaling pathway genes in the YO were two to four orders of magnitude higher in blocked animals compared to controls. These data suggest that increased MIH and CHH synthesis in the ESG contributes to the prevention of molt induction by MLA in blocked animals. The up-regulation of MIH signaling genes in the YO of blocked animals suggests that the YO is more sensitive to MIH produced in the ESG, as well as MIH produced in brain and TG of ESA animals. Both the up-regulation of MIH signaling genes in the YO and of Gl-MIH and Gl-CHH in the ESG, TG, and brain appear to contribute to some G. lateralis individuals being refractory to MLA and ESA.

First evidence of neurons in the male copulatory organ of a spider (Arachnida, Araneae).

Spider males have evolved a remarkable way of transferring sperm by using a modified part of their pedipalps, the so-called palpal organ. The palpal organ is ontogenetically derived from tarsal claws; however, no nerves, sensory organs or muscles have been detected in the palpal bulb so far, suggesting that the spider male copulatory organ is numb and sensorily blind. Here, we document the presence of neurons and a nerve inside the male palpal organ of a spider for the first time. Several neurons that are located in the embolus are attached to the surrounding cuticle where stresses and strains lead to a deformation (stretching) of the palpal cuticle on a local scale, suggesting a putative proprioreceptive function. Consequently, the male copulatory organ of this species is not just a numb structure but likely able to directly perceive sensory input during sperm transfer. In addition, we identified two glands in the palpal organ, one of which is located in the embolus (embolus gland). The embolus gland appears to be directly innervated, which could allow for rapid modulation of secretory activity. Thus, we hypothesize that the transferred seminal fluid can be modulated to influence female processes.

Alleged silk spigots on tarantula feet: electron microscopy reveals sensory innervation, no silk.

Several studies on tarantulas have claimed that their tarsi could secrete fine silk threads which would provide additional safety lines for maintaining a secure foot-hold on smooth vertical surfaces. This interpretation was seriously questioned by behavioral experiments, and more recently morphological evidence indicated that the alleged spigots ("ribbed hairs") were not secretory but most likely sensory hairs (chemoreceptors). However, since fine structural studies were lacking, the sensory nature was not proven convincingly. By using transmission electron microscopy we here present clear evidence that these "ribbed hairs" contain many dendrites inside the hair lumen - as is the case in the well-known contact chemoreceptors of spiders and insects. For comparison, we also studied the fine structure of regular silk spigots on the spinnerets and found them distinctly different from sensory hairs. Finally, histological studies of a tarantula tarsus did not reveal any silk glands, which, by contrast, are easily found within the spinnerets. In conclusion, the alleged presence of silk spigots on tarantula feet is refuted.

Morphology and histology of Lyonet's gland of the tropical tasar silkworm, Antheraea mylitta.

The morphology and histology of Lyonet's gland in the second to fifth instar larvae of Antheraea mylitta Drury (Lepidoptera: Saturniidae) are described. Each of the paired silk glands of this silk worm were associated with a Lyonet's gland. The paired Lyonet's glands were located on the ventrolateral sides of the esophagus, close to the subesophageal ganglion. Whole mount and SEM observations revealed that each Lyonet's gland consisted of a rosette of glandular mass, and a short narrow tubular duct opening into the anterior part of the silk gland (ASG), close to the common excretory duct. In each instar, these glands were unequal in size. The glandular mass was innervated by fine nerves from the subesophageal ganglion, suggesting a neural control for the glandular activity. The glandular mass was made up of clustered long cells wrapped by a thin basal lamina, which was continuous over the non-secretory low columnar cells of the Lyonet's gland duct and ASG. The narrow bases of long cells of each glandular mass led into the lumen of the duct of the gland. Histochemical analysis of fully developed Lyonet's gland showed clustered lipid granules in the gland cells.

Autonomic control of glands and secretion: a comparative view.

The autonomic nervous system together with circulating and local hormones control secretion from glands. This article summarizes histochemical and functional studies on the autonomic innervation and control of secretory glands in non-mammalian vertebrates, including secretion of saliva in the mouth and gastric acid in the stomach, secretion of enzymes and bicarbonate from the pancreas and gut wall, secretion of mucus in the gut epithelium and onto the skin, and salt secretion from salt glands and rectal glands. Cholinergic and adrenergic nerves, directly or indirectly, in combination with different types of peptidergic and other nerves appear to innervate gland tissues and affect secretion in all investigated species.

BDNF promotes target innervation of Xenopus mandibular trigeminal axons in vivo.

BACKGROUND: Trigeminal nerves consist of ophthalmic, maxillary, and mandibular branches that project to distinct regions of the facial epidermis. In Xenopus embryos, the mandibular branch of the trigeminal nerve extends toward and innervates the cement gland in the anterior facial epithelium. The cement gland has previously been proposed to provide a short-range chemoattractive signal to promote target innervation by mandibular trigeminal axons. Brain derived neurotrophic factor, BDNF is known to stimulate axon outgrowth and branching. The goal of this study is to determine whether BDNF functions as the proposed target recognition signal in the Xenopus cement gland. RESULTS: We found that the cement gland is enriched in BDNF mRNA transcripts compared to the other neurotrophins NT3 and NT4 during mandibular trigeminal nerve innervation. BDNF knockdown in Xenopus embryos or specifically in cement glands resulted in the failure of mandibular trigeminal axons to arborise or grow into the cement gland. BDNF expressed ectodermal grafts, when positioned in place of the cement gland, promoted local trigeminal axon arborisation in vivo. CONCLUSION: BDNF is necessary locally to promote end stage target innervation of trigeminal axons in vivo, suggesting that BDNF functions as a short-range signal that stimulates mandibular trigeminal axon arborisation and growth into the cement gland.

Neurturin signalling via GFRalpha2 is essential for innervation of glandular but not muscle targets of sacral parasympathetic ganglion neurons.

Neurturin, a member of the glial cell-derived neurotrophic factor familys of ligands, is important for development of many cranial parasympathetic ganglion neurons. We have investigated the sacral component of the parasympathetic nervous system in mice with gene deletions for neurturin or its preferred receptor, GFRalpha2. Disruption of neurturin signalling decreased cholinergic VIP innervation to the mucosa of the reproductive organs, but not to the smooth muscle layers of these organs or to the urinary bladder. Thus, neurturin and its receptor are involved in parasympathetic innervation of a select group of pelvic visceral tissues. In contrast, noradrenergic innervation was not affected by the gene ablations. The epithelium of reproductive organs from knockout animals was atrophied, indicating that cholinergic innervation may be important for the maintenance of normal structure. Cholinergic neurons express GFRalpha2 on their terminals and somata, indicating they can respond to neurotrophic support, and their somata are smaller when neurturin signalling is disrupted. Colocalisation studies showed that many peripheral glia express GFRalpha2 although its role in these cells is yet to be determined. Our results indicate that neurturin, acting through GFRalpha2, is essential for parasympathetic innervation of the mucosae of reproductive organs, as well as for maintenance of a broader group of sacral parasympathetic neurons.

Motor control of airway goblet cells and glands.

Activation of nerves increases airway mucus secretion. The mucus derives from submucosal glands and epithelial goblet cells. Depending upon species and airway level, innervation comprises parasympathetic (cholinergic), sympathetic (adrenergic) and 'sensory-efferent' pathways. In all species studied, cholinergic mechanisms predominate, particularly in human airways. Muscarinic M3 receptors on the secretory cells mediate the cholinergic response. Tachykinins (substance P and neurokinin A) mediate the sensory-efferent response, acting via tachykinin NK1 receptors. Endogenous mechanisms regulate the magnitude of neurogenic secretion, including enzymes (degrade neurotransmitters), nitric oxide (NO) and vasoactive intestinal peptide (VIP) (regulate stimulated secretion), and muscarinic M2 autoreceptors (inhibit acetylcholine release). Exogenous opioids also inhibit neurogenic secretion prejunctionally. Both VIP and opioids act by opening large conductance, calcium-activated potassium (BK(Ca)) channels. Present understanding of neural control of mucus secretion in animal airways requires translation into human data. This information should lead to rational development of drugs for bronchial diseases in which neurogenic mucus hypersecretion contributes to pathophysiology, including chronic bronchitis and asthma.

[Electron microscopy studies of innervation of nasal mucosa glands in humans]. / Elektronenmikroskopische Untersuchungen zur Innervation der Drüsen der Nasenschleimhaut des Menschen.

BACKGROUND: Seromucous glands are one of the main components of human nasal mucosa. The innervation pattern is important to understand the control of the different physiological glandular functions. In addition to light-microscopical findings electronmicroscopic investigations were performed to get more detailed information on the innervation of nasal glands. PATIENTS AND METHODS: Tissue samples of 16 human inferior turbinates were taken during nasal surgery and preserved in Unicryl or 3.5% phosphate-buffered glutaraldehyde. After fixation ultrathin sections were cut. Electron microscopical structures were photodocumented by using a transmission-electron microscope (EM 902 A Zeiss). RESULTS: Few axons were found in the periglandular tissue. No myo- or glandular-neural tight junctions could be identified. Unmyelinated nerve fibers showed typical components such as neurofilaments, neurotubules and mitochondria in their cytoplasm. An additional control of the glandular secretion by the vascular tone of the fenestrated capillary vessels will be discussed. CONCLUSIONS: Based on these ultramorphological findings further immunoelectron microscopical investigations will follow to demonstrate the various neurotransmitters and their distribution in periglandular axons.

A calcium/calmodulin-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate in the CNS of the cuttlefish Sepia officinalis: localization in specific neural pathways controlling the inking system.

Chemical, biochemical, and immunohistochemical evidence is reported demonstrating the presence in the brain of the cuttlefish Sepia officinalis of a Ca2+-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate, occurring in neurons and fibers functionally related to the inking system. Nitric oxide synthase activity was concentrated for the most part in the cytosolic fraction and was masked by other citrulline-forming enzyme(s). The labile nitric oxide synthase could be partially purified by ammonium sulfate precipitation of tissue extracts, followed by affinity chromatography on 2',5'-ADP-agarose and calmodulin-agarose. The resulting activity, immunolabeled at 150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by antibodies to rat neuronal nitric oxide synthase, depended on NADPH and tetrahydro-L-biopterin, and was inhibited by N(G)-nitro-L-arginine. NMDAR2/3 subunit-immunoreactive proteins migrating at 170 kDa could also be detected in brain extracts, along with glutamate (whole brain: 0.32 +/- 0.03 micromol of glutamate/mg of protein; optic lobes: 0.22 +/- 0.04; vertical complex: 0.65 +/- 0.06; basal lobes: 0.58 +/- 0.04; brachial lobe: 0.77 +/- 0.06; pedal lobe: 1.04 +/- 0.08; palliovisceral lobe: 0.86 +/- 0.05). Incubation of intact brains with 1.5 mM glutamate or NMDA or the nitric oxide donor 2-(N,N-diethylamino)diazenolate-2-oxide caused a fivefold rise in the levels of cyclic GMP, indicating operation of the glutamate-nitric oxide-cyclic GMP signaling pathway. Immunohistochemical mapping of Sepia CNS showed specific localization of nitric oxide synthase-like and NMDAR2/3-like immunoreactivities in the lateroventral palliovisceral lobe, the visceral lobe, and the pallial and visceral nerves, as well as in the sphincters and wall of the ink sac.

Vasoactive intestinal polypeptide (VIP) innervation of the human eyelid glands.

This study was conducted to obtain morphological proof of innervating nerve fibres in the glands of the human eyelid (accessory lacrimal glands of Wolfring, meibomian glands, goblet cells, glands of Zeis, glands of Moll, sweat glands, glands of lanugo hair follicles) and identification of the secretomotorically active neuropeptide vasoactive intestinal polypeptide (VIP) as a common transmitter. Epoxy-embedded ultrathin sections of tissue samples from human eyelids were studied using electron microscopy. Paraffin sections fixed in Bouin-Hollande solution were immunostained with rabbit antiserum against VIP. With the electron microscope we were able to identify nerves in the glandular stroma of all the glands examined with the exception of goblet cells. Intraepithelial single axons were only seen in the parenchyma of Wolfring glands. The morphological findings corresponded with the immunological finding of VIP-positive, nerve-like structures in the same locations, with the exception of lanugo hair follicle glands, and goblet cells. Our findings indicate that the glands of the eyelids and main lacrimal gland represent a functional unit with VIP as a possible common stimulating factor.

Immunohistochemical localization of delta opioid receptors in peripheral tissues.

The distribution of delta opioid receptor (DOR) immunoreactivity (ir) was examined in various peripheral tissues of Sprague-Dawley rats and macaque monkeys, including glabrous and hairy skin, corneas, eyelids, and the lip. DOR-ir was observed in all tissues examined. In addition to the presence of DOR-immunoreactive fibers in subcutaneous nerve bundles and the papillary dermis, we report the existence of positively labeled fibers and terminals in close association with peripheral structures not traditionally assigned a primarily nociceptive function, such as hair follicles, glandular apparatus, and blood vessels. In every case, staining was restricted to small-diameter axons that appeared to terminate as free nerve endings. To further classify DOR-immunoreactive fibers, we examined the extent of colocalization between DOR and three commonly used neuronal subtype markers; tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), and RT-97, a monoclonal antibody which preferentially labels neurons with myelinated axons. Additional double-labeling experiments using the nonspecific neuronal marker Protein Gene Product 9.5 were performed in glabrous skin to determine the percentage of total fiber count that displayed DOR-ir. No colocalization was observed between DOR and RT-97, indicating that DOR-ir is localized to unmyelinated axons. In addition, DOR colocalized with CGRP, but did not colocalize with TH. Taken together, these data support the hypothesis that delta opioid receptors in peripheral tissues are associated with sensory fibers, but not with the terminals of postganglionic sympathetic neurons.

The foam production system of the male Japanese quail: characterization of structure and function.

The research described here characterizes a unique neuromuscular system involved in reproductive behavior--the foam production system of the male Japanese quail (Coturnix japonica). Male quail produce a large amount of foam that is transferred to the female during copulation, enhancing male fertilization success. The source is the foam gland complex, a large sexually dimorphic, androgen sensitive, external protuberance of the dorsal cloaca, consisting of glandular units interdigitated with striated muscle fibers of the sphincter cloacae muscle (mSC). Electromyographic (EMG) analysis of mSC activity in freely moving males interacting with females revealed different characteristics of the EMG signal during copulation, voiding of excreta, and other mSC movement. The amount of mSC activity and also the amount of foam produced were greatly increased by the presence of a female behind a screen. Denervation of mSC eliminated normal mSC movement and also abolished foam production, confirming that mSC activity is the mechanism for foam production. The spinal cord locations of the motoneurons innervating the major cloacal muscles, including mSC, were determined by injecting cholera-toxin conjugated horseradish peroxidase into each muscle. Labelled somata with multiple primary dendrites were located in Area IX of the lateral motor column of synsacral segments 7, 8, or 9 or 8, 9, and 10. The motoneurons serving mSC were intermingled with those projecting to the other cloacal muscles, but there were differences in the rostralcaudal placement of these neural populations. Thus mSC activity is an integral part of the male's reproductive behavior, mSC activity can be socially stimulated, and mSC activity occurring in anticipation of copulation is likely to be functionally significant. Continued investigation of this highly accessible system has the potential to shed light on the mechanisms by which complex motor acts are produced and hormonally regulated.

Increased VIP-positive nerve fibers in the mucous glands of subjects with chronic bronchitis.

The presence and distribution of neuropeptide-containing nerves within bronchial surgical specimens has been investigated in bronchitic (n = 12) and in nonbronchitic subjects (n = 7). Lung tissue, obtained from patients undergoing thoracotomy for limited lung lesions, was processed immediately and analyzed for nerves using the streptavidin-biotin complex peroxidase method with antisera to the neural marker protein gene product 9.5 (PGP 9.5) and the neuropeptides vasoactive intestinal peptide (VIP), substance P (SP), calcitonin-gene related peptide (CGRP). There were no significant differences between the two groups with respect to the density of PGP 9.5-, SP-, or CGRP-positive nerves in both the locations assessed (smooth muscle layer and glands). The density of VIP-positive nerves was significantly higher in the glands of bronchitic than in nonbronchitic subjects. A negative relationship was found between the presence of airway inflammation, as indexed by mononuclear cell tissue infiltration, and the density of PGP 9.5-positive nerves in both smooth muscle and glands. Likewise, a relationship was found between the smoking history (packs/yr and age of onset of smoking) and the density of VIP-positive nerves in glands. These findings support a role for VIP in the hallmark of chronic bronchitis, i.e., sputum production.

Control of venom production and secretion by sympathetic outflow in the snake Bothrops jararaca.

Many studies have examined the morphological and biochemical changes in the secretory epithelium of snake venom glands after a bite or milking. However, the mechanisms of venom production and secretion are not yet well understood. The present study was undertaken to evaluate the role of the sympathetic nervous system in the control of venom production and secretion. Venom glands were obtained from Bothrops jararaca (Viperidae) snakes, either unmilked previously or milked 4, 7 or 15 days before they were killed. Levels of tyrosine-hydroxylase-like immunoreactivity were higher in venom glands collected 4 days after milking, coinciding with the maximal synthetic activity of the secretory cells. The only catecholamine detected by high-performance liquid chromatography was noradrenaline, indicating the presence of noradrenergic fibres in these glands. In reserpine-treated milked snakes, no venom could be collected, and electron microscopic analysis showed narrow rough endoplasmic reticulum cisternae, instead of wide cisternae, and less well-developed Golgi apparatus compared with milked untreated snakes, indicating impairment of protein synthesis and secretion. The administration of isoprenaline or phenylephrine (beta- and alpha-adrenoceptor agonists, respectively) to reserpine-treated milked snakes promoted the widening of the rough endoplasmic reticulum and restored venom production, but only phenylephrine restored the development of the Golgi apparatus and the formation of many secretory vesicles. These results provide the first evidence that the sympathetic nervous system plays an important role in venom production and secretion in the venom glands of Bothrops jararaca. Understanding the importance of noradrenergic stimulation in venom production may provide new insights for research into the treatment of snakebites.

Selective innervation of different target tissues in guinea-pig cranial exocrine glands by sub-populations of parasympathetic and sympathetic neurons.

This study has used multiple-labelling immunohistochemistry and quantitative analysis to examine the projections of subpopulations of parasympathetic and sympathetic neurons to different vascular and secretory structures in five cranial exocrine glands of guinea-pigs. Multiple subpopulations of parasympathetic axons, identified by immunoreactivity (IR) for various combinations of peptides, innervated arteries, arterioles, ducts and acini in sublingual, submandibular, parotid, lacrimal and zygomatic glands, although axons were absent from ducts in the parotid gland. Most parasympathetic axons contained IR for vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), with or without enkephalin (Enk). The proportion of parasympathetic axons that contained Enk-IR varied greatly between target tissues and glands: Enk-IR was more common in axons supplying secretory ducts, acini and arterioles than in axons innervating more proximal arteries; Enk-IR was less common in axons supplying the lacrimal gland than axons supplying the submandibular, lacrimal and zygomatic glands. Sympathetic axons with IR for tyrosine hydroxylase (TH) innervated arterial vessels in all glands, but innervated secretory structures only in the salivary glands. Sympathetic axons supplying proximal arterial segments often contained NPY-IR and sometimes also contained IR for dynorphin. Dynorphin-IR was more common in axons in the parotid, lacrimal and zygomatic glands than in the sublingual and submandibular glands. In contrast, axons supplying arterioles, ducts and acini lacked peptide IR. These results indicate that neuronal pathways regulating proximal arteries in cranial exocrine glands are different from the neuronal pathways regulating arterioles and acini, and may be different from neurons projecting to proximal secretory ducts. Furthermore, the peptides enkephalin, NPY and dynorphin are likely to make variable contributions to autonomic neurotransmission in different arterial segments and in different cranial exocrine glands.

Autonomic and sensory innervation of cat molar gland and blood vessels in the lower lip, gingiva and cheek.

Innervation of the molar gland and blood vessels in the lower lip, gingiva and cheek mucous membrane was investigated in the cat with the aid of whole mount acetylthiocholinesterase (WATChE) histochemistry and retrograde neuronal tracing methods with horseradish peroxidase (HRP) and HRP-conjugated wheat germ agglutinin (WGA-HRP). The molar gland was found to be supplied from the buccal nerve and branches of the mylohyoid nerve on the basis of microdissection of WATChE-stained mandibular preparations under a dissecting microscope. The rostral half of the lower lip-gingiva was innervated by mental branches from the inferior alveolar nerve. The caudal half of the lower lip-gingiva and cheek mucous membrane were observed to be supplied from the buccal nerve. Following injections of HRP/WGA-HRP into the molar gland, lower lip-gingiva and cheek, many retrogradely labeled ganglion neurons were observed in the ipsilateral main and accessory otic ganglia, superior cervical ganglion and mandibular division of the trigeminal ganglion. In the pterygopalatine ganglion, a small number of positive neurons were found, but in a few cases in which the injected tracer was restricted to the lower lip-gingiva and anterior half of the molar gland, labeled neurons were not detected in the main ganglion nor in its accessory microganglia. These findings indicate that the cat molar gland receives a postganglionic parasympathetic supply from the otic ganglia, postganglionic sympathetic input from the superior cervical ganglion and sensory innervation from the trigeminal ganglion by way of the buccal nerve and mylohyoid nerve. Vessels in the rostral half of the lower lip-gingiva receive the same inputs from the inferior alveolar nerve, and vessels in the caudal half receive inputs from the buccal nerve. The vessels in the cheek mucous membrane receive dual parasympathetic supplies from the otic ganglia and the pterygopalatine ganglion by way of the buccal nerve.

Neuroendocrine cell-axonal complexes in the minor vestibular gland.

OBJECTIVE: To study neuroendocrine cells and nerves in vestibular glands of patients with vestibulitis. STUDY DESIGN: Substance P chromogranin, serotonin and neuron-specific enolase immunoreactivity was studied in vestibular gland tissue. Electron microscopic analysis was performed on vestibular duct tissue. RESULTS: Neuroendocrine cells containing immunoreactive serotonin and chromogranin were demonstrated in a vestibular gland duct of a patient with vestibulitis. Substance P immunoreactive axons were found in submucosal nerves, and rare, positively staining axons were seen in the duct epithelium. In another patient neuroendocrine cells with closely applied intraepithelial axons were demonstrated by electron microscopy in the duct of a vestibular gland. CONCLUSION: Intraepithelial axons are present in ducts of vestibular glands, where they are closely applied to serotonin-containing neuroendocrine cells. This close association and the presence of substance P immunoreactivity in local nerves may play a role in the symptomatology of vestibulitis.

Regulation of mucous acinar exocrine secretion with age.

Denny and co-workers (Navazesh et al., 1992) recently reported decreased concentrations of MG1 and MG2 mucins in resting and stimulated whole human saliva with age. The current study was therefore conducted to examine whether there is a corresponding attenuation with age in stimulus secretion coupling regulating mucous cell exocrine secretion. We utilized an in vitro model system, isolated rat sublingual acini, to evaluate the regulation of mucous cell exocrine secretion. Rat sublingual glands are similar to human sublingual and minor mucous glands, both histologically and in terms of their pattern of innervation, which is predominantly parasympathetic. Mucin secretion is thus activated primarily by muscarinic cholinergic agonist and to a lesser extent by vasoactive intestinal peptide (VIP), which is co-localized with acetylcholine in parasympathetic nerve terminals. We isolated sublingual mucous acini from five-month-old and 24-month-old rats and compared the concentration responses for mucin secretion induced by VIP and the muscarinic agonist, arecaidine propargyl ester (APE). Concentration-response curves for VIP were nearly identical for mucous acini from the five-month-old and 24-month-old animals. Values for basal secretion, maximal secretion, and EC50 (approximately equal to 200 nmol/L VIP) were statistically equivalent between both age groups. Concentration-response curves for APE were also very similar between age groups, with no statistically significant difference in basal secretion or EC50 values (approximately equal to 50 nmol/L APE). Maximal secretion was slightly less but statistically different for 24-month-old vs. five-month-old animals, 158% vs. 169% above basal secretion, respectively. Collectively, we found no substantial age-related changes in the secretory responsiveness of salivary mucous cells.