Distribution of FMRFamide-like immunoreactivity in the alimentary tract and hindgut ganglia of the barnacle Balanus amphitrite (Cirripedia, Crustacea)

In this study, the presence and distribution of FMRFamide-like immunoreactivity in the alimentary tract of barnacle Balanus amphitrite were investigated. A net of nerve fibers strongly immunoreactive to FMRFamide-like molecules was localized in the posterior midgut and hindgut. Positive varicose nerve terminals were also localized close to the circular muscle cells and, in the hindgut, close to the radial muscular fibers. Besides this nerve fibers network, one pair of contralateral ganglia was localized in the hindgut, each of them constituted by two strongly FMRFamide-labeled neurons and one nonlabeled neuron. Their immunoreactive axons directed toward the hindgut and posterior midgut suggest an involvement of FMRFamide-like substances in adult B. amphitrite gut motility. The hindgut associated ganglia of barnacles seem to correspond to the terminal abdominal ganglia of the other crustaceans. Since they are the only residual gut ganglia in the barnacle's reduced nervous system, we can hypothesize that gut motility needs a nervous system regulation partially independent of the central nervous system.     

Fine structure of the pinealopetal innervation of the mammalian pineal gland.

The mammalian pineal gland is innervated by peripheral sympathetic and parasympathetic nerve fibers as well as by nerve fibers originating in the central nervous system (central innervation). The perikarya of the sympathetic fibers are located in the superior cervical ganglia, while the fibers terminate in boutons containing small granular vesicles and a few large granular vesicles. Both noradrenaline and neuropeptide Y are contained in these neurons. The parasympathetic fibers originate from perikarya in the pterygopalatine ganglia. The neuropeptides, vasoactive intestinal peptide and peptide histidine isoleucine, are present in these fibers, the boutons of which contain small clear transmitter vesicles and larger granular vesicles. The fibers of the central innervation originate predominantly from perikarya located in hypothalamic and limbic forebrain structures as well as from perikarya in the optic system. These fibers terminate in boutons containing small clear and, in certain fibers, an abundant number of large granular vesicles. In rodents, the majority of the central fibers terminate in the deep pineal gland and the pineal stalk. From these areas impulses might be transmitted further caudally to the superficial pineal gland via neuronal structures or processes from pinealocytes. Several hypothalamic neuropeptides and monoamines might be contained in the central fibers. The intrapineal nerve fibers are located both in the perivascular spaces and intraparenchymally. The majority of the intraparenchymally located fibers terminate freely between the pinealocytes. However, some nerve terminals make synaptic contacts with the pinealocytes and in some species with intrapineal neurons. In fetal mammals, sympathetic, parasympathetic, and central fibers are also present. In addition, an unpaired nerve, connecting the caudal part of the pineal gland with the extreme rostral part of the mesencephalon, is present. This nerve is a homologue to the pineal nerve (nervus pinealis) observed in lower vertebrates.     

The ultrastructure of neuronal-pinealocytic interconnections in the monkey pineal.

Recent ultrastructural studies of neuronal-pinealocytic interconnections in the monkey pineal are reviewed. The pinealocytes in the adult monkey show almost all of the cytological specializations known in subprimate mammals. Adjacent pinealocytes are functionally coupled through ribbon synapses on cell bodies and gap junctions on cell bodies and cell processes. The pinealocytes receive direct synpatic contacts of nerve fibers with cholinergic terminal morphology. Nerve cells restricted to the central portion of the pineal receive synaptic contacts with more than three different morphologically defined types of nerve terminals. In addition to nerve terminals containing small clear vesicles or vesicles of pleomorphic morphology, a pinealocyte's terminal process containing the synaptic ribbon forms a true synaptic contact on the nerve cell body. The diversity of synapses on these nerve cells strongly suggests multiple origins of these neurons rather than a single peripheral parasympathetic origin. The possible involvement of pineal neurons in an intrinsic circuit that regulates the function of pinealocytes and integrates the neural input from the central as well as the peripheral nervous systems is discussed.     

G‐protein alpha subunits distribution in the cyprid of Balanus amphitrite (=Amphibalanus amphitrite) (Cirripedia,Crustacea)

The acorn barnacle Balanus amphitrite is a marine crustacean with six nauplius and one cyprid larval stages and a sessile adult, that represent one of the main constituents of sea biofouling. The cyprid is the last larval stage, specialized for settlement, and the study of its biology is interesting also in the frame of antifouling strategies. In this study, a novel approach to the neurobiology of B. amphitrite cyprid has undertaken, studying immunohistochemically the distribution of some G‐protein α subunits (Gαs, Gαo Gαi, and Gαq) on B. amphitrite cyprid. Gαs‐like immunoreactivity was observed in the intestinal mucosa, oral cone, epithelial cells along the outer face of the mantle and thorax; Gαo into the fibers of the neuropile of the central nervous system; Gαi in oil cells, epithelial cells, and limbs and thorax muscles; Gαq was not detected. The results suggest the involvement of the G‐protein α subunits in different tissues and functions that seem to be in agreement with the distribution of the ones from the same class of G‐proteins in vertebrates. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

Immunolocalization of S100-like protein in the brain of an emerging model organism: Nothobranchius furzeri

The S100 protein in nervous tissue appears to play important roles in regulating neuronal differentiation, glial proliferation, plasticity, development, axonal growth, and in neurogenetic processes. In fish, the adult neurogenic activity is much higher than in mammals. In this study, the localization of S100 protein was investigated in the brain of annual teleost fish, Nothobranchius furzeri, which is an emerging model organism for aging research. By immunohistochemical techniques, S100 immunoreactivity (IR) was detected in glial cells, small neurons, and fibers throughout all regions of central nervous system (CNS) with different pattern of distribution. In the telencephalon, S100 IR was seen in the olfactory bulbs and in different areas of the telencephalic hemispheres. In the diencephalon, S100 positivity was observed in the habenular nuclei of the epithalamus, in the cortical thalamic nucleus, in the dorsal, ventral and caudal portions, the latter with the posterior recessus nucleus, and in the diffuse inferior lobe of the hypothalamus, along the diencephalic ventricle and in the dorsal optic tract. In the mesencephalon, S100 IR was observed in the longitudinal tori, in the optic tectum, and along the mesencephalic ventricle. In the rhombencephalon, S100 IR was shown in valvula and body of the cerebellum, and in some nuclei of the medulla oblongata. The results suggest that S100 may play a key role in the maintenance of the CNS and in neurogenesis processes in the adulthood.     

Development and neuronal dependence of cutaneous sensory nerve formations: Lessons from neurotrophins

Null mutations of genes from the NGF family of NTs and their receptors (NTRs) lead to loss/reduction of specific neurons in sensory ganglia; conversely, cutaneous overexpression of NTs results in skin hyperinnervation and increase or no changes in the number of sensory neurons innervating the skin. These neuronal changes are paralleled with loss of specific types of sensory nerve formations in the skin. Therefore, mice carrying mutations in NT or NTR genes represent an ideal model to identify the neuronal dependence of each type of cutaneous sensory nerve ending from a concrete subtype of sensory neuron, since the development, maintenance, and structural integrity of sensory nerve formations depend upon sensory neurons. Results obtained from these mouse strains suggest that TrkA positive neurons are connected to intraepithelial nerve fibers and other sensory nerve formations depending from C and Aδ nerve fibers; the neurons expressing TrkB and responding to BDNF and NT‐4 innervate Meissner corpuscles, a subpopulation of Merkell cells, some mechanoreceptors of the piloneural complex, and the Ruffini's corpuscles; finally, a subpopulation of neurons, which are responsive to NT‐3, support postnatal survival of some intraepithelial nerve fibers and Merkel cells in addition to the muscle mechanoreceptors. On the other hand, changes in NTs and NTRs affect the structure of non‐nervous structures of the skin and are at the basis of several cutaneous pathologies. This review is an update about the role of NTs and NTRs in the maintenance of normal cutaneous innervation and maintenance of skin integrity. Microsc. Res. Tech. 2010. © 2009 Wiley‐Liss, Inc.     

Neurotrophins and their receptors in the primary olfactory neuraxis

The primary olfactory pathway is an elegant and simple system in which to study neurogenesis and neuronal plasticity because of the simple fact that olfactory receptor neurons (ORNs) are continually generated throughout the adult lifetimes of vertebrates. Thus, neuronal birth, differentiation, survival, axon pathfinding, target recognition, synapse formation, and cell death are developmental events that can be examined in the mature olfactory epithelium (OE). Neurotrophins (nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3, and 4/5) are a family of bioactive peptides that exert their effects by interacting with high- and low-affinity receptors on the surfaces of responsive cells, and have been implicated in several stages of neuronal development throughout the central and peripheral nervous system (CNS and PNS). There has been significant interest within the olfactory community as to how these multifunctional peptides might regulate the cycle of degeneration and regeneration of olfactory receptor neurons. The focus of this review is to highlight what is known about the actions of neurotrophins in the primary olfactory pathway, and to pinpoint future directions that will enable us to further understand their role in olfactory receptor neuron development and turnover.     

Physiological functions of FMRFamide-like peptides (FLPs) in crustaceans

Neuropeptides play important roles in chemical signalling in the central and peripheral nervous systems. One of the largest families of neuropeptides is that of the FMRFamide-like peptides (FLPs). This paper reviews what is known about the physiological functions of FLPs in crustaceans, focussing on the cardiovascular, digestive and neuromuscular systems.     

Biochemical and morphological differentiation of acetylcholinesterase-positive efferent fibers in the mouse cochlea

We have compared the biochemical expression of cholinergic enzymes with the morphological differentiation of efferent nerve fibers and endings in the cochlea of the postnatally developing mouse. Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) are present in the newborn cochlea at specific activities 63% and 25%, respectively, of their mature levels. The relative increases in ChAT, in AChE, and in its molecular forms over the newborn values start about day 4 and reach maturity by about day 10. The biochemical results correlate well with the massive presence of nerve fibers stained immunocytochemically for ChAT and AChE or enzymatically for AChE in the inner and outer hair cell regions. Ultrastructural studies, however, indicate the presence of only few vesiculated fibers and endings in the inner and outer hair cell regions. The appearance of large, cytologically mature endings occurs only toward the end of the third postnatal week. The discrepancy may be resolved in the electron microscope using the enzymatic staining for AChE. Labeling is seen on many nonvesiculated fibers and endings in the hair cell regions, suggesting that the majority of the efferent fibers in the perinatal organ may be biochemically differentiated but morphologically immature. The results may imply that the efferents to inner and outer hair cells develop earlier than indicated by previous ultrastructural studies. Moreover, the pattern of development suggests that in the cochlea, as in other tissues, the biochemical differentiation of the efferent innervation may precede the morphological maturation.     

Peptides in frog brain areas processing visual information

Vision is the most important sensory modality to anurans and a great deal of work in terms of hodological, physiological, and behavioral studies has been devoted to the visual system. The aim of this account is to survey data about the distribution of peptides in primary (lateral geniculate complex, pretectum, tectum) and secondary (striatum, anterodorsal and anteroventral tegmental nuclei, isthmic nucleus) visual relay centers. The emphasis is on general traits but interspecies variations are also noted. The smallest amount of peptide-containing neuronal elements was found in the lateral geniculate complex, where primarily nerve fibers showed immunostaining. All peptides found in the lateral geniculate complex, except two, occurred in the pretectum together with four other peptides. A large number of neurons showing intense neuropeptide thyrosine-like immunoreactivity was characteristic here. The mesencephalic tectum was the richest in peptide-like immunoreactive neuronal elements. Almost all peptides investigated were present mainly in fibers, but 9 peptides were found also in cells. The immunoreactive fibers show a complicated overlapping laminar arrangement. Cholecystokinin octapeptide, enkephalins, neuropeptide tyrosine, and substance P (not discussed here) gave the most prominent immunoreactivity. Several peptides also occur in the tectum of fishes, reptiles, birds, and mammals. Peptides in various combinations were found in the striatum, the anterodorsal- and anteroventral tegmental nucleus, and the isthmic nucleus that receive projections from the primary visual centers. The functional significance of peptides in visual information processing is not known. The only exception is neuropeptide tyrosine, which was found to be inhibitory on retinotectal synapses.     

Innervation of the carotid body: Immunohistochemical,denervation, and retrograde tracing studies

This review presents information about multiple neurochemical substances in the carotid body. Nerve fibers around blood vessels and glomus cells within the chemoreceptive organ contain immunoreactivities (IR) for tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), substance P (SP), galanin (GAL), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), calretinin (CR), calbindin D-28k (CB), parvalbumin (PV), and nitric oxide synthase (NOS). Parasympathetic neurons scattered around the carotid body contain VIP, choline acetyltransferase, and vanilloid receptor 1-like receptor. In the mammalian carotid body, transection of the carotid sinus nerve (CSN) causes the absence or decrease of CGRP-, SP-, and NOS-immunoreactive (IR) nerve fibers, whereas all NPY-IR nerve fibers disappear after removal of the superior cervical ganglion. Most VIP-IR nerve fibers disappear but a few persist after sympathetic ganglionectomy. In addition, the CSN transection appears to cause the acquisition of GAL-IR in originally immunonegative glomus cells and nerve fibers within the rat carotid body. On the other hand, 4%, 25%, 17%, and less than 1% of petrosal neurons retrogradely labeled from the rat CSN contain TH-, CGRP-, SP-, and VIP-IR, respectively. In the chicken carotid body, many CGRP- and SP-IR nerve fibers disappear after vagus nerve transection or nodose ganglionectomy. GAL-, NPY-, and VIP-IR nerve fibers mostly disappear after removal of the 14th cervical ganglion of the sympathetic trunk. The origin and functional significance of the various neurochemical substances present in the carotid body is discussed.     

Anatomical organization of retinotopic motion-sensitive pathways in the optic lobes of flies

Anatomical methods have identified conserved neuronal morphologies and synaptic relationships among small-field retinotopic neurons in insect optic lobes. These conserved cell shapes occur across many species of dipteran insects and are also shared by Lepidoptera and Hymenoptera. The suggestion that such conserved neurons should participate in motion computing circuits finds support from intracellular recordings as well as older studies that used radioactive deoxyglucose labeling to reveal strata with motion-specific activity in an achromatic neuropil called the lobula plate. While intracellular recordings provide detailed information about the motion-sensitive or motion-selective responses of identified neurons, a full understanding of how arrangements of identified neurons compute and integrate information about visual motion will come from a multidisciplinary approach that includes morphological circuit analysis, the use of genetic mutants that exhibit specific deficits in motion processing, and biomimetic models. The latter must be based on the organization and connections of real neurons, yet provide output properties similar to those of more traditional theoretical models based on behavioral observations that date from the 1950s. Microsc. Res. Tech. 62:132-150, 2003.     

Innervation of the gastric mucosa

A plethora of neuronal messengers ("classical" transmitters, gaseous messengers, amino acid transmitters, and neuropeptides) are capable of mediating or modulating gastric functions. Accordingly, the stomach is richly innervated. Gastric nerves are either intrinsic to the gastric wall, i.e., they have their cell bodies in the intramural ganglia and thus belong to the enteric nervous system, or they reach the stomach from outside, originating in the brainstem, in sympathetic ganglia, or in sensory ganglia. Topographically, the nerve fibers in the stomach reach all layers from the most superficial portions of the gastric glands to the outer smooth muscle layer. This wide distribution implies that virtually all different cell types may be reached by neuronal messengers. Within the gastric mucosa endocrine and paracrine cells (e.g., gastrin cells, ECL cells, somatostatin cells), exocrine cells (parietal cells, chief cells, mucous cells), smooth muscle cells, and stromal cells are regulated by neuronal messengers. The sensory innervation, responding to capsaicin, plays an important role in mucosal protection, and in ulcer healing. Presumably also other nerves are involved and a plasticity in the neuropeptide expression has been demonstrated at the margin of gastric ulcers. Taken together, available data indicate a complex interplay between hormones, paracrine messengers and neuronal messengers, growth factors and cytokines in the regulation of gastric mucosal activities such as secretion, local blood flow, growth, and restitution after damage.     

New ways to look at axons in Caenorhabditis elegans

In the nematode Caenorhabditis elegans, a well-established model organism for the analysis of nervous system development and function, nerve processes can be labelled in the intact animal with markers based on the "Green Fluorescent Protein" (GFP). The generation of GFP variants with improved brightness and modified emission spectra potentiated the use of this marker for in vivo labelling of subcellular structures. This made it possible to label different groups of neurons and their axons in the same animal with GFP variants of different spectral characteristics. Here I show with double labelling experiments that spatial relationships of axons in small axon bundles can now be resolved at the light microscopic level. In the future this will largely circumvent the need for time-consuming electron microscopic reconstructions to detect local defects in axon outgrowth. Furthermore, I demonstrate that neuronal processes can now be traced even in the head ganglia, an area of the nervous system that was previously almost inaccessible for analysis due to the compact arrangement of cell bodies and axons.     

Development of histamine-immunoreactivity in the Central nervous system of the two locust species Schistocerca gregaria and Locusta migratoria

Locusts are attractive model preparations for cellular investigations of neurodevelopment. In this study, we investigate the immunocytochemical localization of histamine in the developing ventral nerve cord of two locust species, Schistocerca gregaria and Locusta migratoria. Histamine is the fast neurotransmitter of photoreceptor neurons in the compound eye of insects, but it is also synthesized in interneurons of the central nervous system. In the locust ventral nerve cord, the pattern of histamine-immunoreactive neurons follows a relatively simple bauplan. The histaminergic system comprises a set of single, ascending projection neurons that are segmentally arranged in almost every neuromere. The neurons send out their axons anteriorly, forming branches and varicosities throughout the adjacent ganglia. In the suboesophageal ganglion, the cell bodies lie in a posteriolateral position. The prothoracic ganglion lacks histaminergic neurons. In the posterior ganglia of the ventral nerve cord, the somata of the histaminergic neurons are ventromedially positioned. Histamine-immunoreactivity starts around 50% of embryonic development in interneurons of the brain. Subsequently, the neurons of the more posterior ganglia of the ventral nerve cord become immunoreactive. From 60% embryonic development, the pattern of soma staining in the nerve cord appears mature. Around 65% of embryonic development, the photoreceptor cells show histamine-immunoreactivity. The histaminergic innervation of the neuropile develops from the central branches toward the periphery of the ganglia and is completed right before hatching.     

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.     

The terminal nerve and its relation with extrabulbar "olfactory" projections: lessons from lampreys and lungfishes

The definition of the terminal nerve has led to considerable confusion and controversy. This review analyzes the current state of knowledge as well as diverging opinions about the existence, components, and definition of terminal nerves or their components, with emphasis on lampreys and lungfishes. I will argue that the historical terminology regarding this cranial nerve embraces a definition of a terminal nerve that is compatible with its existence in all vertebrate species. This review further summarizes classical and more recent anatomical, developmental, neurochemical, and molecular evidence suggesting that a multitude of terminalis cell types, not only those expressing gonadotropin-releasing hormone, migrate various distances into the forebrain. This results in numerous morphological and neurochemically distinct phenotypes of neurons, with a continuum spanning from olfactory receptor-like neurons in the olfactory epithelium to typical large ganglion cells that accompany the classical olfactory projections. These cell bodies may lose their peripheral connections with the olfactory epithelium, and their central projections or cell bodies may enter the forebrain at several locations. Since "olfactory" marker proteins can be expressed in bona fide nervus terminalis cells, so-called extrabulbar "olfactory" projections may be a collection of disguised nervus terminalis components. If we do not allow this pleiomorphic collection of nerves to be considered within a terminal nerve framework, then the only alternative is to invent a highly species- and stage-specific, and, ultimately, thoroughly confusing nomenclature for neurons and nerve fibers that associate with the olfactory nerve and forebrain.     

The distribution of histamine and serotonin in the barnacle's nervous system

The use of antisera directed against conjugates of histamine and serotonin has revealed the locations of neurons labeling for these transmitters in the nervous system of barnacles. Photoreceptors label for histamine but not serotonin and also satisfy a number of other criteria indicating that histamine is their neurotransmitter. Photoreceptors also take up radioactively labeled histamine but not serotonin. Within the barnacle's brain no somata are consistently found that label with antiserum against histamine, but one to three pairs of small cells, depending on species, label with antiserum against serotonin. The most impressive serotonin-like immunoreactivity in the brain, however, is in a pair of large fibers ascending through the circumesophageal connectives and ramifying extensively. Within the ventral ganglion, the only other ganglion in the barnacle, ten pairs of cells label with antiserum against histamine. These neurons are confined to the posterior portion of the ganglion but ramify extensively throughout the ganglion. Antiserum against serotonin labels about 15 cell pairs, depending on species, located throughout the ganglion. The positions of the arbors of many of these cells suggest that these amines have a role in modulating either the motor pathways underlying feeding or the visual pathways responsible for the detection of shadows.     

Distribution of neurotrophin and TrK receptor-like immunoreactivity in the adrenal gland of birds

The occurrence and localization of neurotrophins and their specific TrK receptor-like proteins in the adrenal gland of chicken, duck and ostrich were examined by immunohistochemical methods. In all species studied NGF-, TrK A- and TrK C-like immunoreactivity was observed in neurons and fibers of adrenal ganglia. Thin TrK A- and TrK C-like immunoreactive fibers were also observed among chromaffin cells. NT-3-like immunoreactivity was detected in chromaffin cells as revealed by the double immunolabelings NT-3/chromogranin A and NT-3/DbetaH. The interrenal tissue never showed IR to any neurotrophins and TrK tested, and none of the adrenal structures displayed immunoreactivity to BDNF and TrK B. Double immunolabelings NGF/TrK A, NGF/TrK C and TrK A/TrK C showed colocalization in some neurons and fibers in adrenal ganglia. In adrenal glands of the species studied, the distribution of neurotrophins and TrK receptors could suggest an involvement of NT-3 on neuronal populations innervating adrenal ganglia by means of its high affinity receptor TrK C and low affinity receptor TrK A. In addition, NGF could be utilized by neuronal populations of adrenal ganglia through its preferential receptor TrK A by an autocrine or paracrine modality of action.