The central consequences of the application of capsaicin to one peripheral nerve in adult rat

This paper reviews the central consequences of local application of capsaicin to one nerve in adult animals. 1) Marked chemical changes occur in the central terminals of C fibres. These include depletion of the enzyme FRAP and the peptides SP, CCK, somatostatin, CGRP and an increase of VIP. Maximal depletions occur if the nerve is soaked with capsaicin solutions with a concentration higher than 3 mM. The depletion begins by 7 days and is complete by 11. Recovery begins at about 110 days and is largely complete by 200. Our studies have concentrated on the effects of 40 mM capsaicin examined 14 days after the application. 2) Capsaicin treatment of a peripheral nerve decreased the ability of C fibres in that nerve to excite or to inhibit spinal cord cells. It produces a marked expansion of receptive fields of some cells in the dorsal horn which respond to A fibre stimulation. It is proposed that this change is not due to anatomical changes but to disinhibition. A further example of receptive field expansion is seen after treatment of the mouse infraorbital nerve which defocuses the normally precise projection of individual whiskers onto single cells in the barrel field of the somatosensory cortex. 3) Behavioural consequences follow the treatment of one adult nerve with capsaicin. In the area subserved by the treated nerve, there is a raised threshold to response to chemical and thermal stimuli, no change in the response to mechanical stimuli and an increase of autotomy following nerve section. 4) The aim of the experiments was to determine the role of C fibres in producing the changes seen in spinal cord following peripheral nerve section. Capsaicin treatment of nerve imitates the central effect of complete nerve section in certain important ways. Both result in a marked expansion of the receptive field of some cells. The effect is produced by a change of chemical transport. The results show that C fibres influence the connection of A fibres onto spinal cord cells.     

Electrophysiological experiments on the mechanism and accuracy of neuromuscular specificity in the axolotl.

The supracoracoideus muscle of the axolotl shoulder girdle is innervated by two nerves, the supracoracoideus nerve (SC) supplying most of the muscle and the posterior supracoracoideus (PSC) supplying the posterior corner. All the muscle fibres are multiply innervated and at the border between the two innervations many muscle fibres, when penetrated by a microelectrode, show junction potentials from both nerves. In such cases one junction potential is often very small, below the threshold for exciting muscle contraction, the other large and effective at exciting the muscle. If the SC nerve is cut, the territory of the PSC nerve expands over several weeks. Upon regrowth of the cut nerve it reinnervates its old muscle fibres and removes the previous foreign innervation, the borderline between the two nerve territories being established exactly as before. This depends upon two processes, sprouting of nerves and a competitive repression of transmission from nerves ending on foreign muscle fibres.     

The relation between nerve fibres and dopamine cells of the ruminant lung

Synopsis A study of the distribution and ultrastructure of the special chromaffin or dopamine cells of the ruminant lung has been made. The results with fluorescence microscopy show that the adult sheep has larger numbers of such cells than has the lamb, calf or adult goat. Contrary to earlier findings, their numbers have been found to be as great in the intralobular tissue as in the pleura, septa and other structures. Electron microscopic examination of the cells shows that they resemble true mast cells; some contain two types of granule, the appearance of which suggests that part of the amine has been discharged. Although in many places the cells are closely associated with adrenergic nerve fibres no evidence was found to suggest that they had any functional relation with one another.     

Axonal sprouts of the hypoglossal nerve implanted in the superior cervical ganglion of adult rats establish synaptic contacts under long-lasting GABA effect. An experimental degeneration study

Experimental degeneration was used in this study to determine if the hypoglossal nerve implanted already in the superior cervical ganglion of adult rat under GABA treatment has established morphologically-identifiable synapses with the dendrites of principal ganglion cells. The implanted hypoglossal nerve trunk was cut in a re-operation, and the ganglionic samples were studied by electron microscopy after 0, 6, 12, 24 and 48 h survival times. First signs of degenerative changes were found in the myelinated and non-myelinated axons alike, 6 h after axotomy. The fine-structural signs of degeneration resembled those of the preganglionic nerve fibres. Degenerating nerve terminals establishing synaptic contacts with the dendrites of the principal ganglion cells were also seen, indicating that the axonal sprouts of the implanted hypoglossal nerve established synaptic contacts with the ganglion cells. It remained, however, to be elucidated whether or not these synapses of the hypoglossal nerve are functionally active contacts while the preganglionic innervation is also present within the ganglion.     

Competence of blastomeres for the expression of molecular tissue markers is acquired by diverse mechanisms in the embryo of Platynereis (Annelida)

This paper is devoted to the role of cell divisions for the establishment of histospecificity in the embryo of the spiralian, Platynereis dumerilii (Annelida). We have incubated successive cleavage stages in cytochalasin B (CCB) and observed whether the cells thereafter were able to acquire the competence for expressing histospecific antigens of larval gland cells (labelled by the monoclonal antibody OI64) and of neural components of the ventral nerve cord (labelled by mAb OI7 or by testing acety1cholinesterase activity), respectively. Incubation in CCB results in permanent cleavage arrest, but does not necessarily interfere with biochemical differentiation of such markers. Synthesis of the differentiation marker specific for larval gland cells does not require any cleavages but this capacity becomes restricted to the 1a and 1b cell lines if cleavages are allowed to occur. In contrast, the progenitors of neural cells need at least 6.5 h of normal development before they acquire the competence to synthesize two neural differentiation markers, which can be demonstrated after at least two more days of development. Thus, prespecifying and diversifying cleavages are a prerequisite for neurogenesis and production of the investigated neural markers. Competence for the expression of histospecific antigens may also depend on cell-cell interactions. If 20–24 h old embryos are treated with puromycin, pioneering fibres fail to grow out from a pair of posterior nerve cell progenitors as they would have done normally 24–48 h after fertilization. Concomitantly, a number of potential nerve cells which now do not come into contact with pioneering fibres do not express the neural antigen. This suggests that a local inductive stimulus from the pioneering fibres normally imprints cell fate onto ventral plate cells and turns them into neuroblasts. Correspondence to: A.W.C. Dorresteijn     

Rapid anterograde and retrograde tracing from mesenteric nerve trunks to the guinea-pig small intestine in vitro

A novel technique for rapid anterograde labelling of cut axons in vitro was used to visualise the peripheral branches of mesenteric nerve trunks supplying the guinea-pig small intestine. Biotinamide, dissolved in an artificial intracellular solution, was applied to the cut ends of the mesenteric nerves and the tissue was maintained in organ culture overnight. Labelled nerve fibres were visualised by fluorescein isothiocyanate (FITC)-conjugated streptavidin. Intense staining of nerve fibres and terminal varicosities in the ganglia and internodal strands of the myenteric plexus was achieved up to 15 mm from the application site. Filled fibres formed baskets around some myenteric nerve cell bodies, suggesting target-specific neurotransmission. When combined with multiple-labelling immunohistochemistry for tyrosine hydroxylase (TH), calcitonin gene-related protein (CGRP) or choline acetyltransferase (ChAT), most anterogradely labelled nerve fibres, and many pericellular baskets, were found to be TH immunoreactive, indicating their postganglionic sympathetic origin. Double-labelling immunohistochemistry revealed that the postganglionic sympathetic pericellular baskets preferentially surrounded 5-hydroxytryptamine (5-HT)-handling myenteric neurons. Some biotinamide-filled fibres were CGRP immunoreactive, and are likely to originate from spinal sensory neurons. We describe for the first time many pericellular baskets labelled from the mesenteric nerves which were ChAT immunoreactive. Retrogradely filled intestinofugal nerve cell bodies were also observed, all of which had a single axon arising from a small nerve cell body with short filamentous or lamellar dendrites. Many of these cells were ChAT immunoreactive. This in vitro technique is effective in identifying the fine arrangement of nerve terminals arising from nerve trunks in the periphery.     

Development of substance P and neurokinin A immunoreactivity in ganglia supplying nerves to the submandibular glands of the rat

Developing submandibular, trigeminal and superior cervical ganglia, which provide innervation to the submandibular glands, were studied for substance P (SP)-and neurokinin A (NKA)-immunoreactive (IR) ganglion cells and nerve fibres in rat. These ganglia were examined by using an indirect immunofluorescence technique at daily intervals from the 16th day in utero (i.u.) until birth, and subsequently on the 2nd, 5th, 7th, 12th, 16th, 30th, 42nd postnatal day and in the adult (3 months). In the submandibular ganglion SP- and NKA-IR cells and fibres first appeared in considerable numbers on the 19th day i.u. (in one sample out of five on the 18th day i.u.), when more than 90% of the ganglion cells were immunoreactive to SP and NKA. The number stayed at more than 90% to the 7th postnatal day and then slowly decreased to the levels of adult animals (18% SP, 17% NKA). The first SP- and NKA-IR ganglion cells and fibres appeared in the trigeminal ganglion on the 18th day i.u. when they represented 7% (SP) and 4% (NKA) of the ganglion cells. The number of SP- and NKA-IR cells increased steadily, reaching a maximum at the time of birth when 68% (SP) and 74% (NKA) of the ganglion cells were immunoreactive. Thereafter they began to decrease toward the level of an adult rat (10% SP, 11% NKA). In the superior cervical ganglion only a few SP-and NKA-IR ganglion cells were detected from the 19th day i.u. to the fifth postnatal day. Positive ganglion cells were also occasionally found in the nerve trunks outside the superior cervical ganglion. From the seventh day onwards no SP- or NKA-IR ganglion cells were found. SP-and NKA-IR SIF (small intensively fluorescent) cells were detected from the 16th postnatal day onwards.     

The peripheral and central changes resulting from cutting or crushing the afferent nerve supply to the whiskers

In neonatal rats, crushing or cutting the infraorbital nerve, the sensory nerve supply to the whiskers, has been found to prevent cortical barrel formation. However, both procedures are followed by regeneration of one-third to one-half of the nerve fibres and reinnervation of the whiskers. By counting fibres in individual whisker follicle nerves, it has been shown that 29-67% (mean 45%) of the myelinated fibres regenerate to the whiskers after a crush compared to 24-56% (mean 39%) after a cut. Further differences between the crush and cut lesions were indicated by studies on the time course of regeneration. Counts of the regenerating fibres at various ages as well as recordings of cortical evoked potentials in normal, nerve-crushed and nerve-cut animals showed that recovery was 3-4 days earlier in the nerve-crushed, compared with the nerve-cut animals. In normal and nerve-crushed animals the evoked potential was first detectable 2-3 days after birth while the response after nerve cut could not be recorded until day 7. Even after 60 days the amplitude of responses on both crushed and cut pathways was only about one-third of normal, while the latency was prolonged (normal 5.8 +/- 0.25 ms, crush 6.5 +/- 0.26 ms, cut 7.7 +/- 0.67 ms). Central changes occurring as a result of nerve cut or crush have been studied by microelectrode recordings from the trigeminal nucleus (the first synaptic level) and the somatosensory cortex. These also indicate clearly the greater severity of the cut lesion. Thus, in crushed animals, all levels of the trigeminal nucleus as well as the cortex show only minor modifications. The whiskers occupy the same total area and responses from all whiskers are present at their normal sites. However, after nerve cut, the responses from both the trigeminal nucleus and cortex show clear abnormalities. The total whisker area is reduced with a concomitant expansion of responses from the nose, check, lower jaw, and whiskers by the eye and ear. In addition, only one-third to one-half of the whiskers give responses. The site of these abnormalities is localized to the trigeminal nucleus since all whiskers show innervation in the peripheral nerve. It is suggested that the longer recovery time as well as the reduced accuracy of reinnervation may contribute to the poorer central recovery after a nerve cut.     

The origin of the aldehyde-fuchsin-negative nerve fibres of the median eminence of the hypophysis: a gonadotropic centre

Summary In a large number of adult female and male specimens of Rana temporaria, the pars ventralis of the tuber cinereum of the hypothalamus was completely isolated from the brain. The pars ventralis retained its normal connections with the hypophysis. The vascular supply of the whole isolated region was left intact. Two and a half months after operation, in some animals, the interruption of the nervous pathways was still complete. In these animals it could be shown that all or nearly all A. F.-negative nerve fibres of the median eminence are axons of nerve cells localized in the area periventricularis of the pars ventralis tuberis. According to our results in Rana, the same A. F.-negative nerve fibres of the higher vertebrates are probably also axons of homologous nuclei of the tuber cinereum.In accordance with previous experiments, it could be concluded that the nerve cells of which the A. F.-negative fibres are axons, form an important centre that, via the pars distalis of the hypophysis, is responsible for the seasonal development of the ovaries, eggs and oviducts in the females and for the seasonal development of the thumb pads in the males.     

猫腰髓白质年龄相关的形态学变化

以青年成年猫(1-3龄,2-2．5 kg)和老年猫(12龄,3-3．5kg)L6段脊髓白质为研究对象,用 神经丝蛋白(NF)免疫染色显示神经纤维,用改良的Holzer结晶紫染色显示所有胶质细胞并用成年动物Golgi 法显示其形态,用胶质纤维酸性蛋白(GFAP)免疫染色显示星形胶质细胞。光镜下对青年猫与老年猫腰髓白质 中神经纤维和胶质细胞进行形态学观察和定量研究。与青年猫相比,老年猫腰髓白质中的神经纤维密度显著下 降(P相似文献   

AXONAL TRANSPORT OF RADIOACTIVITY IN THE GOLDFISH OPTIC SYSTEM FOLLOWING INTRAOCULAR INJECTION OF LABELLED RNA PRECURSORS

After injection of the tritiated RNA precursors [³H]guanosine, [³H]uridine or [³H]orotic acid into the eye of goldfish, labelled TCA-soluble material and RNA appeared to be axonally transported to the contralateral optic tectum. From the time courses of arrival in the tectum,‘average’rates of transport of 6 mm/day for the soluble material and 1·7 mm/day for the RNA were calculated. If the optic nerve was cut after the transported material had arrived in the tectum, about 60 per cent of the TCA-soluble material disappeared by 7 days after the cut, but almost none of the RNA. After a further 8- to 13-day period, the TCA-soluble material had declined by a further 50 per cent from the 7-day value, but the RNA by only 20 per cent. Thus, relatively little RNA was lost when the optic axons degenerated, an observation which suggested that the RNA might be extra-axonal. However, if the optic nerve was crushed before the arrival of the transported material, RNA did not appear in the tectum until the regenerating optic nerve fibres arrived. Therefore, the presence of RNA must be dependent on intact nerve fibres. Moreover, in the earliest stages of regeneration the proportion of transported RNA to TCA-soluble material was considerably higher than normal, suggesting that the regenerating fibres arrived in the tectum already carrying RNA. This implies that the RNA itself was transported in the optic fibres.     

Induction of action potentials in frog slow muscle fibres paralysed by alpha -bungarotoxin

Slow muscle fibres in the frog are normally incapable of generating action potentials. However, several days after an intramuscular injection of alpha - bungarotoxin, they acquire the ability to generate action potentials. It appears that alpha -bungarotoxin induces the action potential mechanism in slow fibres because it blocks acetylcholine receptors, and thus interferes with the action of non-quantal acetylcholine leaking from nerve terminals, or because the toxin has some other, as yet undefined, action on nerve or muscle.     

Chemosensory integration in the spiny lobster: Ascending activity in the olfactory-globular tract

Summary In the frog,Rana esculenta, when the influence of the efferent vestibular system was eliminated, the spontaneous activity of single afferent fibres recorded from one branch of the nerve of the horizontal semicircular canal (HC) or of the nerve of the vertical anterior canal (VAC) was inhibited in 16–17% of the cases when stimulating electrically the other branch of the same ampullary nerve. Moreover, the spontaneous activity of about 200 afferent fibres was recorded from the nerves of the HC and VAC in three experimental situations. In the first one, the brain was destroyed, or the left vestibular nerve cut as it enters the brain stem, and all the branches of the left vestibular nerve were cut except for the one recorded (VAC or HC nerve); in the second one, recordings were made on the peripheral end of the ampullary nerve previously cut near the ampulla; in the third situation they were made on the ampullary nerve after having cut the vestibular nerve between the periphery and Scarpa's ganglion close to Scarpa's ganglion. Statistical comparisons of the distribution of the spontaneous frequencies and of the mean activities between the experimental situations show that the activities were greater in the second or third experimental situations than in the first one. These results could be explained by the existence of an inhibitory feedback loop outside the brain including Scarpa's ganglion and mediated by receptor-receptor fibres.Abbreviations HC horizontal semicircular canal - PE peripheral end of the ampullary nerve - VAC vertical anterior semicircular canal This research was supported by a grant from D.G.R.S.T. (Aide à la Recherche n 77.7.1127)     

Frontal deafferentation of the mediobasal hypothalamus in the neonatal rat and its effects on the preoptico-infundibular LHRH-tract

Summary By use of the peroxidase-antiperoxidase-complex (PAP) immunohistological method, the preoptico-infundibular LHRH-tract was studied in adult female rats in which frontal hypothalamic deafferentation was performed at the third or tenth postnatal day. In the former group, this LHRH-tract appeared to be similar to that of the intact controls; the animals showed regular vaginal cycles and ova were present in their oviducts. In the latter group, however, marked reduction in the number of the LHRH-nerve fibers was observed behind the sites of the deafferentation in the mediobasal hypothalamus (MBH), whereas LHRH-immunoreactive perikarya and nerve fibers containing the immunoreactive material were seen rostral to the plane of severance. In these animals reduction of LHRH-fibers in the MBH was accompanied by an anovulatory syndrome characterized by constant vaginal cornification and polyfollicular ovaries. Comparing the glial scar formation induced by the cut, significant differences were detected between the two experimental groups. In the animals deafferented on the 3rd day of life, reduction of nerve cells was seen along the cut, but LHRH-fibers crossing the thin glial scar were detectable in large numbers. On the other hand, in the animals deafferented on the 10th postnatal day, extensive glial scar tissue appeared to interrupt the LHRH-fibers rostral to the cut.     

The early development of retinal ganglion cells with uncrossed axons in the mouse: retinal position and axonal course

The carbocyanine dye, DiI, has been used to study the retinal origin of the uncrossed retinofugal component of the mouse and to show the course taken by these fibres through the optic nerve and chiasm during development. Optic axons first arrive at the chiasm at embryonic day 13 (E13) but do not cross the midline until E14. After this stage, fibres taking an uncrossed course can be selectively labelled by unilateral tract implants of DiI. The earliest ipsilaterally projecting ganglion cells are located in the dorsal central retina. The first sign of the adult pattern of distribution of ganglion cells with uncrossed axons located mainly in the ventrotemporal retina is seen on embryonic day 16.5, thus showing that the adult line of decussation forms early in development. A small number of labelled cells continue to be found in nasal and dorsal retina at all later stages. At early stages (E14-15), retrogradely labelled uncrossed fibres are found in virtually all fascicles of the developing nerve, intermingling with crossed axons throughout the length of the nerve. At later stages of development (E16-17), although uncrossed fibres pass predominantly within the temporal part of the stalk, they remain intermingled with crossed axons. A significant number of uncrossed axons also lie within the nasal part of the optic stalk. The position of uncrossed fibres throughout the nerve in the later developmental stages is comparable to that seen in the adult rodent (Baker and Jeffery, 1989). The distribution of uncrossed axons thus indicates that positional cues are not sufficient to account for the choice made by axons when they reach the optic chiasm.     

Substance P and neurokinin A immunoreactive nerve fibres in the developing salivary glands of the rat

The time of appearance and distribution of substance P (SP) and neurokinin A (NKA) immunoreactive nerve fibres in developing salivary glands of the rat were studied by the use of indirect immunohistochemical methods. The glands were examined at daily intervals from the 15th day in utero (i.u.) until birth, and subsequently on the 2nd, 5th, 7th, 12th, 16th and 30th postnatal day. The findings were compared to samples from adult. The first SP- and NKA-immunoreactive (IR) nerve fibres appeared on the 19th day i.u. in the parotid and submandibular glands and were abundantly distributed around developing ductal branches. In the mesenchyme around the developing ductal branches of the parotid gland the fibres appeared on the 20th day i.u. In the submandibular gland NKA-IR fibres appeared in the mesenchyme surrounding the developing ductal branches on the 19th day i.u. and SP-IR fibres on the 21st day i.u. Around blood vessels of both glands, SP- and NKA-IR fibres made their appearance only much later, on the second postnatal day. The number of SP- and NKA-IR nerve fibres in the developing salivary glands was already high on the 19th day i.u. when they were first detected. From this point up to the 16th postnatal day the glands were richly innervated by the fibres, but later the numbers slowly decreased to adult levels. The abundance of SP- and NKA-IR nerve fibres especially around the ductal branches and secretory structures in the developing salivary glands suggests a role in the functional maturation of salivary glands.     

Substance P and neurokinin A immunoreactive nerve fibres in the developing salivary glands of the rat.

The time of appearance and distribution of substance P (SP) and neurokinin A (NKA) immunoreactive nerve fibres in developing salivary glands of the rat were studied by the use of indirect immunohistochemical methods. The glands were examined at daily intervals from the 15th day in utero (i.u.) until birth, and subsequently on the 2nd, 5th, 7th, 12th, 16th and 30th postnatal day. The findings were compared to samples from adult. The first SP- and NKA-immunoreactive (IR) nerve fibres appeared on the 19th day i.u. in the parotid and submandibular glands and were abundantly distributed around developing ductal branches. In the mesenchyme around the developing ductal branches of the parotid gland the fibres appeared on the 20th day i.u. In the submandibular gland NKA-IR fibres appeared in the mesenchyme surrounding the developing ductal branches on the 19th day i.u. and SP-IR fibres on the 21st day i.u. Around blood vessels of both glands, SP- and NKA-IR fibres made their appearance only much later, on the second postnatal day. The number of SP- and NKA-IR nerve fibres in the developing salivary glands was already high on the 19th day i.u. when they were first detected. From this point up to the 16th postnatal day the glands were richly innervated by the fibres, but later the numbers slowly decreased to adult levels. The abundance of SP- and NKA-IR nerve fibres especially around the ductal branches and secretory structures in the developing salivary glands suggests a role in the functional maturation of salivary glands.     

The neurohypophysis of the crested newt

Summary In the median eminence of the newt a medial region and two lateral regions are described.In cross section, the medial region appears to be made up of 1) an outer or glandular zone (Zone I) containing aldehyde-thionine-positive and negative nerve fibres and blood capillaries. Nerve fibres appear aligned in palisade array along the capillaries. 2) An inner zone (Zone II) made up of a) a layer of aldehyde-thionine-positive nerve fibres (fibrous layer) belonging to the preoptic hypophyseal tract and b) a layer of ependymal cells lining the infundibular lumen and reaching the blood vessels with their long processes.The lateral regions display a less pronounced stratification and aldehyde-thionine positive nerve fibres are nearly absent.A slender lamina (ependymal border) containing mainly aldehyde-thionine-positive nerve fibres and ependymal cells connects the median eminence to the pars nervosa.At the ultrastructural level, in the outer zone of the medial region at least 4 types of nerve fibres and nerve endings are identified:Type I nerve fibres containing granular vesicles of 700–1000 Å and clear vesicles (250–400 Å).Type II nerve fibres containing granular vesicles and polymorphous granules of 900–1300 Å and clear vesicles (250–400 Å).Type III nerve fibres containing dense granules of 1200–2000 Å and clear vesicles of 250–400 Å.Type IV nerve fibres containing only clear vesicles of 250–400 Å. In the inner zone too, all these nerve fiber types are found among ependymal cells, while the fibrous layer consists of nerve fibres containing granules of 1200–2000 Å in diameter.In the lateral regions Type I, Type II and Type IV nerve fibres and their respective perivascular terminals are found; axons containing dense granules (1200–2000 Å) are scanty. In these regions typical synapses between Type I nerve fibres and processes rich in microtubules are visible.The classification and functional significance of nerve fibres in the median eminence are still unsolved, but it may be assumed that nerve fibres of the medial region belong to both the preoptic hypophyseal and tubero hypophyseal tract, while the lateral regions are characterized by nerve fibres of the tubero hypophyseal tract. Peculiar specializations of the ependymal cells in the median eminence of the newt are also discussed.Work supported by a grant from the Consiglio Nazionale delle Ricerche.The authors are indebted to Mr. G. Gendusa and P. Balbi for technical assistance.     

Some morphological aspects of the innervation of a spider's silk gland

A structure in the ampullate silk gland of an orb-weaving spider, previously thought to be a pressure receptor, was studied by electron microscopy to clarify its sensory function. This lamellated 'bulbous region' is situated between the gland and the beginning duct, which consists of a layered, acellular matrix (muco-protein), surrounded by a flat epithelium and partly covered by connective tissue. The epithelial cells are densely filled by a mesh-work of microtubules and interconnected by extensive cell junctions, both indicating exposure to mechanical stress. The few nerve fibres found to supply the bulbous region do not exhibit specialized dendritic endings and provide no evidence for a receptor function. The abundance of translucent vesicles of 400-500 A (synaptic'?) and dense granules of 1000-1500 A (neurosecretory?) indicate efferent fibres, possibly controlling the secretion of the matrix. The function of the bulbous region remains to be investigated. Similar nerve fibres but with distinct synaptic foci were noted in the adjacent gland duct in large numbers. The neuro-glandular synapses have a characteristic location, namely, opposing the mesaxon-like invagination of the epidermal cells. These nerve fibres are believed to play a role in water uptake and/or secretion of the extra-cellular duct lining.