Neurosecretory Fibers Research Articles

Microstructural and ultrastructural study of pituitary in highfin grouper Cromileptes altivelis

Pituitary was one of most complicated endocrine gland,the function of it was diversification.Pi-tuitary could modulate the activities of the body organs and tissues.It was necessary to investigate structure of fish pituitary in order to provide the basis for histological morphology,reproductive biology and pathology.Using improved staining,Mallory and PAS staining method,and transmission electron microscopy technique,the microstructure and the ultrastructure of the pituitary of Cromileptes altivelis was examined.Results showed that: the pituitary of the C.altivelis was a solid spherical glands,located in the ventral diencephalon,and was composed of neurohypophysis and adenohypophysis.According to the shape and size of excretive granule contained in nerve fibers,the neurosecretory fibers could be differentiated into A1,A2 and B-type.Two types of pituitary cells could be observed in the neurohypophysis,I-type pituitary cells and II-type pi-tuitary cells.Adenohypophysis consistsed of three parts: rostral pars distails(RPD),proximal pars distalis(PPD) and pars intermedia(PI).It could be identified six endocrine cells in adenohypophysis.Adrenocorticotrophic(ATCH) cells and prolactin(PRL) cells existed in the RPD,growth hormone(GH) cells,thyrotrophs(TSH) cells and gonadotrophic hormone(GTH) cells could be found in the PPD,only one endocrine cell were observed,it was melanophore-stimulating hormone(MSH) cells.And one non-secretory cell type-stellate cells(SC) could also be seen in adenohypophysis.The sort of gonadotrophic hormone cells and the composition and distribution of endocrine cell of adenohypophysis were discussed.There was only one kind of GTH cells,but they had many forms.The endoplasmic reticulum of some GTH cells significantly expanded,foam based,and it relatively had a few secretory granules in the cytoplasm.Since the release of secretion of some GTH cells,nuclear was smaller,degenerate,or even disassemble,and secretory granules was preternaturally rich in the cytoplasm.There also had many GTH cells that were not found vacuole,and the edge of cell showed lace-shaped or notch,mitochondria well-developed.

Morphology and histology of pituitary gland of torrential stream fish Nemacheilus mooreh (sykes)

Pituitary lies ventral to the brain behind optic chiasma in the depression of parasphenoid bone attached to the floor of the mid ventral infundibulum. It is Leptobasic type and show transitional condition between caudo and dorsobasic type, sella turcica, a depression provides space to lodge pitutary with thick connective tissue septum. The pituitary gland is composed of nervous part, the neurohypophysis, and a glandular part, the adenohypophysis. The neurohypophysis develops from the infundibular floor and provides a stalk to pituitary. The distal part of neurohypophysis is brightly stained due to presence of neuro secretory material. The positively stained neurosecretory fibres extensively innervate the pars intermedia and rostral pars distalis. Adenohypophysis develops from the roof of the stomodaeum and is divisible into pars intermedia and pars distalis. Pars distalis is composed of a proximal part containing deeply stained cyanophils and acidophils. The rostral and proximal pars distalis and pars intermedia are arranged in a linear fashion with the rostral pars distalis at the anterior extremity. In N. mooreh, pars intermedia completely surround the distal region of neurohypophysis. The study will provide a deeper insight into morphohistology and cell types of pituitary gland of the freshwater Homalopterid teleost.

ULTRASTRUCTURE OF THE DISCONNECTED PARS NERVOSA OF THE CRESTED NEWT (TRITURUS CRISTATUS CARNIFEX LAUR)

SUMMARY The morphological changes in the pars nervosa of the crested newt caused by transection of the hypothalamic floor just in front of the median eminence, were studied at various postoperative intervals. One to 2 days after the operation some dense bodies can be observed within the neurosecretory fibres. However, the most conspicuous changes are detectable after 4 to 7 days. Neurosecretory fibres, appear heavily electrondense as the elementary granules become more densely packed. The fibres often break into fragments and appear widely separated from one another, modifying the general pattern of the pars nervosa. Both ependymal cells and pituicytes are characterized by a strong macrophagic activity towards degenerating nerve fibres which often appear to be engulfed by them. Myelin-like formations containing vacuoles, empty vesicles and dense granules (1200–2000 A in diameter) can be observed among the nerve fibres. Pituicytes, generally rare in the normal animal, show an apparent increase in their num...

Cyclic Changes In The Hypothalamo Neurohypophysial System of Xenentodon Cancila (HAM.)

The presence of functional hypothalamo-hypophysial neurosecretory system in vertebrates has led to revise earlier views regarding the hypothalamus-pituitary relationship. The hypothalamo-neurohypophysial system which consists of hypothalamic nuclei, their axonal fibres forming tractus preoptico-hypophyses and the neurohypophysis) works as a morphological as well as a physiological connection between the hypothalamus and the pituitary gland. In fishes, fibres from the neurosecretory hypothalamic nuclei terminate in the neurohypophysis, which remains inter digitized with the adenohypophysis and provides a very close association between the neurosecretory fibres and pars intermedia of the pituitary gland forming a neuro-intermediate lobe. Hypothalamo-Neurosecretory complex of Xenentodon cancila consists of Nucleus Pre-opticus (NPO), Nucleus Lateralis Tuberis (NLT) and their axonal tracts. NPO is a paired structure situated on either side of the third ventricle anterodorsal to the optic chiasma. NPO is divisible into a dorsal Pars Magnocellularis (PMC) consisting of large neurosecretory cells and Pars Parvocellularis (PPC) formed of smaller neurons. Neurons of PMC and PPC contribute beaded axons to form neurohypophysial tract. Herring bodies are seen in the anterior as well as posterior neurohypophysis.

Intravenous Inoculation of a Bat-Associated Rabies Virus Causes Lethal Encephalopathy in Mice through Invasion of the Brain via Neurosecretory Hypothalamic Fibers

The majority of rabies virus (RV) infections are caused by bites or scratches from rabid carnivores or bats. Usually, RV utilizes the retrograde transport within the neuronal network to spread from the infection site to the central nervous system (CNS) where it replicates in neuronal somata and infects other neurons via trans-synaptic spread. We speculate that in addition to the neuronal transport of the virus, hematogenous spread from the site of infection directly to the brain after accidental spill over into the vascular system might represent an alternative way for RV to invade the CNS. So far, it is unknown whether hematogenous spread has any relevance in RV pathogenesis. To determine whether certain RV variants might have the capacity to invade the CNS from the periphery via hematogenous spread, we infected mice either intramuscularly (i.m.) or intravenously (i.v.) with the dog-associated RV DOG4 or the silver-haired bat-associated RV SB. In addition to monitoring the progression of clinical signs of rabies we used immunohistochemistry and quantitative reverse transcription polymerase chain reaction (qRT-PCR) to follow the spread of the virus from the infection site to the brain. In contrast to i.m. infection where both variants caused a lethal encephalopathy, only i.v. infection with SB resulted in the development of a lethal infection. While qRT-PCR did not reveal major differences in virus loads in spinal cord or brain at different times after i.m. or i.v. infection of SB, immunohistochemical analysis showed that only i.v. administered SB directly infected the forebrain. The earliest affected regions were those hypothalamic nuclei, which are connected by neurosecretory fibers to the circumventricular organs neurohypophysis and median eminence. Our data suggest that hematogenous spread of SB can lead to a fatal encephalopathy through direct retrograde invasion of the CNS at the neurovascular interface of the hypothalamus-hypophysis system. This alternative mode of virus spread has implications for the post exposure prophylaxis of rabies, particularly with silver-haired bat-associated RV.

Intravenous inoculation of silver-haired bat rabies virus, but not of a canine strain, elicits lethal encephalophathy in mice by fast brain invasion via neurosecretory hypothalamic fibers

Neurotropic rabies virus (RV) is transmitted by saliva, most often through the bite of an infected canine, which leads to an always fatal encephalopathy by invasion of the CNS through nerve fibers innervating the affected muscle. In contrast, transfer by bat bites or scratches, the most frequent cause of human rabies in the USA, introduces RV in rather low amounts intradermally. In both scenarios, RV has also access – in addition to nerves – to lymph and blood. However, the effects of this vascularly distributed share of the viral inoculum have never been examined. Our study aimed to elucidate if RV circulating in the vascular system is able to directly invade the brain and if this postulated route is strain dependent. Furthermore, we wanted to identify putative entry ports by which hematogenously spread RV preferentially gains access to the brain.

The fine structural organization of sternal glands of pseudergates and workers in termites (Isoptera): A comparative survey

Thirty-nine species belonging to different families of termites are studied to give a comprehensive view of the evolution of the sternal glands. Several modifications occurring at cuticular and cytological levels are described in neuter castes. The outer epicuticle is always pierced by epicuticular pores. In advanced termites the epicuticular filaments greatly increase in number and length creating a thick layer. The pore canals gradually enlarge while the cuticle changes into a lattice structure lining an extracellular space in which the secretion is stored. Two classes of cells are present in basal termites (Mastotermitidae, Hodotermitidae, Termopsidae and Kalotermitidae) but their glandular structures greatly differ between families. A more complex organization with three classes of cells is found in the Serritermitidae and Rhinotermitidae. A regressive evolution occurs in the Termitidae where only two classes of cells are present. A dual nervous control (campaniform sensilla and neurosecretory fibers) is found in lower termites, except for the Hodotermitidae which have mechanosensory bristles. In the other families, neurosecretory fibers are lacking. A comparison with phylogenetic data is given. A more versatile role of sternal glands in neuter castes is hypothesized.

New Insights on Saccus vasculosus Evolution: A Developmental and Immunohistochemical Study in Elasmobranchs

The saccus vasculosus (SV) is a circumventricular organ of the hypothalamus of many jawed fishes whose functions have not yet been clarified. It is a vascularized neuroepithelium that consists of coronet cells, cerebrospinal fluid-contacting (CSF-c) neurons and supporting cells. To assess the organization, development and evolution of the SV, the expression of glial fibrillary acidic protein (GFAP) and the neuronal markers γ-aminobutyric acid (GABA), glutamic acid decarboxylase (GAD; the GABA synthesizing enzyme), neuropeptide Y (NPY), neurophysin II (NPH), tyrosine hydroxylase (TH; the rate-limiting catecholamine-synthesizing enzyme) and serotonin (5-HT), were investigated by immunohistochemistry in developing and adult sharks. Coronet cells showed GFAP immunoreactivity from embryos at stage 31 to adults, indicating a glial nature. GABAergic CSF-c neurons were evidenced just when the primordium of the SV becomes detectable (at stage 29). Double immunolabeling revealed colocalization of NPY and GAD in these cells. Some CSF-c cells showed TH immunoreactivity in postembryonic stages. Saccofugal GABAergic fibers formed a defined SV tract from the stage 30 and scattered neurosecretory (NPH-immunoreactive) and monoaminergic (5-HT- and TH-immunoreactive) saccopetal fibers were first detected at stages 31 and 32, respectively. The early differentiation of GABAergic neurons and the presence of a conspicuous GABAergic saccofugal system are shared by elasmobranch and teleosts (trout), suggesting that GABA plays a key function in the SV circuitry. Monoaminergic structures have not been reported in the SV of bony fishes, and were probably acquired secondarily in sharks. The existence of saccopetal monoaminergic and neurosecretory fibers reveals reciprocal connections between the SV and hypothalamic structures which have not been previously detected in teleosts.

Co-localization of neuronal nitric oxide synthase with arginine–vasotocin in the preoptic–hypothalamo–hypophyseal system of the teleost Oreochromis niloticus

This study provides evidence that, in the preoptic–hypothalamo–hypophysial system of the teleost Oreochromis niloticus, several sub-populations of arginine–vasotocin (AVT)-producing neurons and neurosecretory fibers terminals express neuronal nitric oxide synthase (nNOS)-like molecules. The co-localization between nNOS and AVT was demonstrated by means of double immunofluorescence staining with the confocal microscope. This study is the first to provide evidence that nNOS may be co-localized with AVT in neurons of a non-mammalian vertebrate.

Electron Microscopy Study of Anterior Neuropituitary of the Sterlet Acipenser ruthenus

Structural organization of sterlet anterior neuropituitary (ANP), a homologue of the median tubercle of Tetrapoda, was studied by method of electron microscopy. The ANP was conventionally divided in three zones in dorsoventral direction. Neurosecretory fibers (NsF) were concentrated mainly in the central and in the external zones. Terminals of NsF are directly adjacent to the basal membrane (BM) of the brain vascular sheath to form a morphological substrate for release of neurohormones into the pericapillary space. The presence of invaginations of the brain connective tissue sheath, which are in a close contact with NsF, indicates an increase of the area of neurosecretory contacts. Neurosecretory granules (NsG) of various shape and size are revealed in NsF and their terminals. Comparison of light-microscopy immunohistochemical data with localization of NsF terminals containing NsG of different diameters allows reconstructing distribution of neurosecretory elements of various ergicity in the ANP. Patterns of co-localization of various biologically active substances were revealed in the same varicose dilatations or terminals. Detection of typical synaptic contacts in the sterlet ANP indicates possibility of the cholinergic pituitary regulation, alongside with the peptide- and monoaminergic regulation.

Efferent innervation of photoreceptors in spiders.

The anterior median (AM) eye of the nocturnal spider Araneus ventricosus showed a marked circadian oscillation of sensitivity, but that of the diurnal spider Menemerus confusus showed no circadian oscillation. The AM eyes of the noct/diurnal spiders Argiope amoena and A. bruennichii have two types of photoreceptor cells with different sensitivities. The more sensitive cells showed a circadian oscillation of sensitivity, but the less sensitive cells did not. The circadian sensitivity change of the eyes was controlled by efferent neurosecretory fibers in the optic nerve. Illuminating the brain increased the frequency of efferent impulses in the optic nerve of Argiope, showing that certain photosensitive neurons are present in the brain. However, it seemed that the cerebral photosensitive neurons may be different from the efferent neurosecretory cells. The response of the cerebral photosensitive neurons increased transiently following diminution of the light intensity striking the eyes. The interaction between the cerebral photosensitive neurons and the eyes seemed to play a role in increasing this response.

Development of the caudal neurosecretory system of the nile tilapia Oreochromis niloticus: an immunohistochemical and electron microscopic study.

The development of the caudal neurosecretory system (CNSS) of the Nile tilapia, Oreochromis niloticus, has been investigated by means of UI/oCRF (urotensin I/ovine corticotropin-releasing factor) immunohistochemistry and transmission electron microscopy. UI-like immunoreactive perikarya and fibers are first detected in the caudal spinal cord of larval fish about 4 days after hatching (stage 21). In the region of the future urophysis two bundles of strongly immunoreactive neurosecretory fibers are observed. At this stage, neurosecretory axons terminate on the meninx sheath of the spinal cord with immature neurosecretory terminals. The histogenesis of the urophysis begins at stage 24. The future neurohemal organ consists of a small ventral swelling of the spinal cord, which is associated with dilated vessels. At this stage, neurosecretory axons terminate on the basal lamina of the ingrowing blood vessels. Further development occurs by means of progressive branching of vessels and the concomitant increase in the number of neurosecretory terminals. In the caudal spinal cord, immunoreactive neurons also increase in number and progressively differentiate morphologically. Typical features of the mature CNSS are recognizable in 4-month-old juveniles. Data suggest that in tilapia both the synthesis and the release of urophysial hormones begin before morphogenesis of the neurohemal organ takes place.

Innervation and control of the adenohypophysis by hypothalamic peptidergic neurons in teleost fishes: EM immunohistochemical evidence.

Previous light microscopic studies have revealed neuropeptide-immunoreactive neurosecretory fibers in the teleostean neurohypophysis, and ultrastructural work has reported direct innervation of endocrine cells by the terminals of fibers penetrating the adenohypophysis. This paper reviews our recent data from ultrastructural, immunohistochemical, receptor localization, and superfusion studies, which suggest a role for neuropeptides in the control of teleost pituitary secretion. We have used a combination of pre- and post-embedding electron microscopic immunolabeling methods to determine which neuropeptides are present in fibers innervating the pituitaries of three species: Poecilia latipinna, Dicentrarchus labrax, and Clarias gariepinus. Numerous axon profiles with immunoreactivity for the neurosecretory peptides vasotocin and isotocin formed large Herring bodies and terminal-like boutons in contact with corticotropic, growth hormone, thyrotropic, and pars intermedia cells. Numerous melanin-concentrating hormone-immunoreactive fibers and scarcer neurotensin and corticotropin-releasing factor-immunoreactive fibers showed similar distributions, terminating close to pars intermedia and corticotropic cells. Somatostatin, cholecystokinin, galanin, substance P, neuropeptide Y, growth hormone-releasing factor, thyrotropin-releasing hormone, and gonadotropin-releasing hormone-immunoreactivities were found in small calibre fibers penetrating among growth hormone, thyrotropic, and gonadotropic cells. These morphological findings have been supplemented by autoradiographic studies, which showed the distribution of binding sites for vasotocin, isotocin, galanin, and neuropeptide Y ligands over specific groups of pituitary cells, and superfusion studies that showed growth hormone release was stimulated by growth hormone-releasing factor and thyrotropin-releasing hormone, but inhibited by somatostatin. The implications of these results for neuropeptidergic control of teleostean pituitary secretions are discussed.

Nitric oxide synthase in the caudal neurosecretory system of the teleost Oreochromis niloticus

This study provides evidence that, within the caudal neurosecretory system of the teleost Oreochromis niloticus, neurons express nitric oxide synthase (NOS)-like molecules. The presence of NOS-like molecules was demonstrated by means of NADPH-diaphorase (NADPHd) staining and NOS immunohistochemistry. In the caudal spinal cord, NOS-positive neurosecretory cell bodies and neurosecretory fibers were observed. In addition, NOS-positive structures were found in the urophysis which correspond to neurosecretory axon terminals. Cellular co-localization of NOS and ovine corticotropin-releasing factor (oCRF) immunoreactivities confirmed that the NOS-positive structures belong to the caudal neurosecretory system. The present results suggest that NO may participate in the caudal neuroendocrine function.

Expression of neural cell adhesion molecules, NCAMs, and their polysialylated forms, PSA-NCAMs, in the developing rat pituitary gland.

Neural cell adhesion molecules (NCAMs) can undergo post-translational modifications, such as the addition of polysialic acid chains, thus generating PSA-NCAMs, which are expressed mainly during development. Since polysialylation considerably modifies NCAM adhesivity, expression of NCAMs and PSA-NCAMs has been investigated in the developing hypophysis by immunohistochemistry. At embryonic day 13 (E13), an antibody against NCAM outlined all cellular profiles in the entire Rathke's pouch; this labelling persisted until adulthood. NCAM expression increased in all lobes during development and concerned all pituitary cell types. In contrast, at E13, PSA-NCAMs were only detected in the neural lobe, solely constituted of pituicytes at this stage, and the tuberal lobe, the only lobe expressing hormonal mRNA at the same stage. PSA-NCAMs expression increased in the neural lobe at E17 with the arrival of the neurosecretory fibres and persisted into adulthood. In the anterior lobe, PSA-NCAMs appeared at E15 where their distribution was similar to that of the differentiating corticotrophic cells; at subsequent stages, their expression extended to the whole anterior lobe. Only two cell types, corticotrophic and somatotrophic cells, remained labelled in the adult gland. In the intermediate lobe, melanotrophic cells never expressed PSA-NCAMs but these were expressed on folliculo-stellate cells at birth, preceding the onset of innervation. These results suggest that NCAMs and PSA-NCAMs play a role in pituitary histogenesis, cell differentiation and neurointermediate lobe innervation.

Immunocytochemical Examination of Catecholaminergic Innervation of Neurosecretory Fibers and Pituicytes in the Rat Neurohypophysis

Immunocytochemical Examination of Catecholaminergic Innervation of Neurosecretory Fibers and Pituicytes in the Rat Neurohypophysis

Neuroarchitecture of the tritocerebrum of Drosophila melanogaster.

The organisation of the tritocerebrum of Drosophila melanogaster was studied by Bodian-Protargol reduced silver staining, Golgi-silver impregnation, horseradish peroxidase (HRP), and cobalt-chloride labelling of neurones and transmission electron microscopy. Nerve fibres of six categories were found to project to the tritocerebrum. (1 and 2) The sensory fibres from the internal mouthpart sensilla known to course along pharyngeal and accessory pharyngeal nerves were found to project in mainly two tiers, in the tritocerebrum. (3) Stomodaeal nerve fibres also project along the pharyngeal nerve, to the tritocerebrum. (4) Cells of the pars intercerebralis (PI) project along the median bundle and arborise in the tritocerebrum. HRP labelling and subsequent examination by transmission electron microscopy indicated their neurosecretory nature. (5 and 6) Two tracts of ascending fibres, designated as dorsal and ventral ascending tracts, were found to project to the tritocerebrum. Some of the sensory fibres from the labial nerve extend close to the sensory projections of the tritocerebrum, suggesting a possible convergence of the two sensory inputs. In the tritocerebrum, the sensory input, the stomodaeal input, the neurosecretory fibres of PI, and the ascending fibres were found to have overlapping fields, suggesting mutual interaction. The medial subesophageal ganglion and the tritocerebrum may interact through the ventral ascending tract.

Reorganization of neural peptidergic systems in the median eminence after hypophysectomy

Earlier studies have shown the formation of a novel neural lobe after hypophysectomy, an experimental manipulation that causes transection of neurohypophyseal nerve fibers and removal of pituitary hormones. The mechanisms that underly this regenerative process are poorly understood. The localization and number of peptide-immunoreactive (-IR) fibers in the median eminence were studied in normal rats and in rats at different times of survival after hypophysectomy using indirect immunofluorescence histochemistry. The number of vasopressin (VP)-IR fibers increased in the external layer of the median eminence in 5 d hypophysectomized rats. Oxytocin (OXY)-IR fibers decreased in the internal layer and progressively extended into the external layer. At long survival times (9 and 16 months) both VP- and OXY-IR fibers had a bilayered distribution occupying both the external and internal layers. Double-labeling experiments combining VP and tyrosine hydroxylase antisera as well as OXY and growth hormone-releasing factor antisera showed that injured neurosecretory fibers growing into the external layer displaced fibers from parvocellular cells originally located there. As a result, there was essentially an inversion in the distribution of these fibers within the median eminence. Galanin (GAL)- and cholecystokinin (CCK)-IR fibers exhibited a similar pattern of distribution after the lesion. Thus, after 5 d there was an increase in GAL- and CCK-IR fibers in the internal layer. At 14 and 30 d, the number of GAL- and CCK-IR fibers progressively decreased, but after longer survivals (9 and 16 months) there was a dramatic reappearance. Dynorphin (DYN)-LI showed a dramatic increase at all levels of the median eminence at short survival times after hypophysectomy, followed by a subsequent decrease to a final stage of a few, strongly immunoreactive fibers in the external layer at longer survival times. Vasoactive intestinal polypeptide (VIP)- and peptide histidine-isoleucine (PHI)-IR fibers in hypophysectomized animals had already contacted portal vessels 5 d after hypophysectomy, and from then on progressively increased in numbers. Finally, most of the peptide fibers described above formed dense innervation patterns around the large blood vessels along the lateral borders of the median eminence. The present results show that hypophysectomy induces a wide variety of changes in hypothalamic neurosecretory fibers. Not only is the expression of several peptides in these fibers modified following different survival times, but a reorganization of the distribution of immunoreactive fibers within the median eminence is demonstrated. The hypothesis is raised that regeneration of injured neurosecretory fibers may be dependent on changes in the expression of peptides possessing trophic actions.

Age-dependent neurosecretion release induced by dopamine in the corpora cardiaca of Blattella germanica (L.) (Dictyoptera : Blattellidae)

In addition to glial cells, intrinsic glandular cells and ordinary axons, the corpora cardiaca of Blattella germanica (L.) (Dictyoptera : Blattellidae) contain 4 types of neurosecretory fibers originating from the brain (types 1, 2, 3, and 4), which can be recognized on the basis of the size, form, and electron density of their neurosecretory granules. A comparative ultrastructural study of the corpora cardiaca from normal females and from dopamine-treated (1 μg) females has been carried out at different stages within the first ovarian cycle (freshly emerged: day 0; 2-day-old: pre-vitellogenesis; 4-day-old: beginning of vitellogenesis; 6-day-old: full vitellogenesis; 8-day-old: post-vitellogenesis, period of ootheca transport). Quantitative data on the exocytotic configurations observed in each type of fiber (1–4) have led to the following conclusions: (a) the exocytotic configurations in control specimens are too infrequent to prompt any inference about the dynamics of neurosecretion release in the different types of fibers; (b) dopamine treatment induces a stimulation of the exocytotis phenomenon, whose extent depends on the specimen age and on the type of fiber. Therefore, on days 2 and 6, dopamine: preferentially stimulates neurosecretion release in the fibers of types 3 and 4 respectively.

Calcitonin gene-related peptide (CGRP)-immunoreactive nerve terminals in the whole mount preparations of the dog urethra.

To visualize the entire shape of the intraepithelial nerve fibers, whole mount preparations of the dog urethra were produced and immunostained with an antiserum against CGRP, one of the predominant substances contained in the nerves. The immunoreactive nerves in the lamina propria were smooth (non-beaded) in appearance and weak in immunoreaction. Within the epithelium, they displayed typical beaded profiles and were intense in immunoreaction. The intraepithelial fibers branched and wound into an extensive network with wide meshes ("reticular terminal"). The bead-like swellings included large ones resembling Herring bodies in hypophyseal neurosecretory fibers. Another type of nerve terminal, consisting of fine and weakly immunopositive fibers, was also found in the epithelium. These branched in dendritic or in dense bouquet-like fashion, occupying smaller areas ("bouquet-like terminals). Vesicular swellings often characterized these terminals, though they were smaller and more uniform in size and far less in their amount of immunoreactive substance than were the swellings in the reticular terminals. Both types of nerve terminals originated from the same nerve trunk. The connection between the reticular and bouquet-like terminals, which may presumably represent secretory and receptive parts, respectively, morphologically supports the possible occurrence of an axon reflex in the urethral CGRP neurons. Our whole mount preparations, when doubly stained with CGRP and serotonin antibodies, further revealed the CGRP-positive reticular terminals being closely associated with serotonin- or CGRP-immunoreactive paraneurons dispersed in the epithelium.