Postembryonic changes in the peripheral electrosensory system of a weakly electric fish: addition of receptor organs with age

The organization of the peripheral electrosensory system of the cheek was studied in an age-graded series of Sternopygus dariensis in Nissl-stained sections and silver-stained whole mounts of skin. As in other gymnotoids, both ampullary and tuberous electroreceptors are present. Small fish have only one ampullary organ or tuberous organ per axon, and the number of receptor organs per axon increases with age in both ampullary and tuberous systems. Large fish may have up to ten tuberous organs per axon, although the distribution of tuberous organs per axon is bimodal with one peak occurring at a single receptor organ per axon and the other peak shifting upward in relation to the age of the fish. The ampullary system adds receptor organs at a faster rate and a large fish may have 20 ampullary organs per axon. With increasing size, the number of sensory receptor cells in each organ remains constant for both types of electroreceptors. Evidence is presented for addition of new electroreceptor units by de novo production in small fish and increases in the number of organs in existing electroreceptor units by division of previously formed organs in medium-sized and large fish. As the surface area of the skin increases with growth, the density of electroreceptor units decreases and, although new receptor organs are still being added to existing receptor units, no generation of new receptor units occurs in medium-sized to large fish.     

Ultrastructural studies of physiologically identified electrosensory afferent synapses in the gymnotiform fish, Eigenmannia

Eigenmannia is a weakly electric fish that emits a constant-frequency electric organ discharge (EOD). Probability coder (P unit) and phase coder (T unit) electroreceptive afferents differentially encode changes in EOD amplitude and phase, respectively. physiologically identified T and P units were intracellularly labelled with HRP and their terminals were examined with electron microscopy to determine their postsynaptic targets. This technique reveals that phase and amplitude are relayed to first-order electrosensory neurons by two parallel but not independent pathways. P-type afferents terminate on granular interneurons, basilar pyramidals, and polymorphic cells, electrosensory lateral line lobe targets that monitor amplitude modulations, but P-type afferents do not contact spherical cells. T-type afferents relay phase information to spherical cells and thus form a separate afferent pathway. T unit terminals do not synapse directly on basilar pyramidal cells. Collateral branches from T-type afferents, however, were also found to terminate on granule and polymorphic cells, thereby adding phase information into the amplitude channel. P- and T-type afferents exhibit cellular specificity by forming synaptic junctions with different subsets of post synaptic targets in the deep neuropil. The afferent terminals make either asymmetric chemical or gap junction synapses depending on the identity of the post synaptic target. T units contacting granule cells or polymorphic cells had not been previously described. Two possible roles of adding phase to amplitude information are discussed in terms of electrolocation.     

Effects of denervation upon receptor cell survival and basal cell proliferation in tuberous electroreceptor organs of a weakly electric fish

Weakly electric fish generate electric fields for the purposes of electrolocation and communication. These fields are detected by specialized receptor organs: the tuberous organs. In the present study we investigated the effects of denervation upon receptor cell survival and progenitor (basal) cell proliferation rate. The left, infraorbital, anterior lateral line nerve of brown ghosts (Apteronotus leptorhynchus) was sectioned, and the proximal stump was dipped in ricin to prevent regrowth. In groups of four, the animals were given two daily injections of the cell proliferation marker bromodeoxyuridine (BrdU) for 2 days at 1, 2, 3, or 4 weeks following denervation. At the completion of the BrdU injection schedule, a piece of cheek skin, rostroventral to the eye, was removed from the left (denervated) and the right (intact) sides and processed for light microscopy or immunocytochemistry. Our results show: (1) there is progressive receptor cell death and tuberous organ degeneration following denervation; (2) basal cell proliferation increases steadily with time after denervation and tuberous organ degeneration; and (3) despite denervation, some proliferating basal cells differentiate into receptor cells, but these new receptor cells eventually die. These results suggest that innervation is essential for tuberous electroreceptor cell survival and that the rate at which basal cells proliferate is regulated by receptor cell health, locally released factors, or both. © 1994 Wiley-Liss, Inc.     

Development of the electrosensory nervous system in Eigenmannia (Gymnotiformes): I. The peripheral nervous system

The nerves of the anterior lateral line system in embryonic and larval stages of the weakly electric gymnotiform fish Eigenmannia were visualized by injection of the fluorescent marker DiI into the primordium of the anterior (ALLN) and posterior (PLLN) lateral line nerves. Examination of developmental series reveals that the nerve fibers that innervate the electrosensory and mechanosensory components of the anterior lateral line system are present before the first mechanoreceptors and electroreceptors have differentiated. This suggests that nerve fibers might induce the formation of lateral line receptors. Whereas the innervation of the mechanoreceptive system is already established at an early stage, the afferent innervation of electroreceptors continues to arborize in the periphery, presumably by following pioneer axon pathways. The earliest recognizable stage of the anterior lateral line nerve ganglion (ALLNG) is evident 2 days after spawning. The ganglion shows two germinal cell masses that develop into the supraorbital-infraorbital and the hyomandibular placodes. The supraorbital-infraorbital placode forms the dorsal part of the ALLNG; the hyomandibular placode forms the ventral part of the ALLNG. Counts of ALLNG cells in embryonic, larval, and adult stages of Eigenmannia show that, at each stage examined, the number of ganglion cells is always significantly larger than the number of mechanoreceptors and electroreceptor units in the periphery. During development, the distribution of ALLNG cell diameters shifts from a unimodal distribution in juveniles to a bimodal distribution in adults, peaking at 8 microns and 18 microns. These results suggest that tuberous electroreceptive organs, which are innervated by the large ALLNG cells, may not be functional prior to day 18. Our results further suggest that the number of ALLNG cells correlates with the rate of induction of lateral line receptors in the periphery.     

Peripheral organization and central projections of the electrosensory nerves in gymnotiform fish

The electrosensory system of weakly electric gymnotiform fish is described from the receptor distribution on the body surface to the termination of the primary afferentsin the posterior lateral line lobe (PLLL). There are two types of electroreceptor(ampullary and tuberous) and a single type of lateral line mechanoreceptor (neuromast). Receptor counts in Apteronotus albifronsshow that (1) neuromasts are distributed as in other teleosts; (2) ampullary receptors number 151 on one side of the head and 208 on one side of the body; (3) tuberous receptors were estimated to number 3,000-3,500 on one side of the head and 3,500-5,000 on one side of the body. The distribution of each receptor type is described. Each receptor is innervated by a single primary afferent. Electro-sensory afferents have myelinated cell bodies in the ganglion of the anterior lateral line nerve (ALLN). The distribution of these ganglion cell diameters is strongly bimodal in Apteronotus and Eigenmannia: The smaller-diameter cells may be those which innervate ampullary electroreceptors, the larger-diameter tuberous electroreceptors. Transganglionic HRP transport techniques were used to determine the first-order connections of the anterior lateral line nerve in six species of gymnotiform fish. Small branches of the ALLN were labeled so as to determine the somatotopic organization in the PLLL. The PLLL is divided into four segments from medial to lateral, termed medial, centromedial, centrolateral, and lateral segments (Heiligenberg and Dye, '81). Representations of the head are found rostrally in each zone, and the trunk is mapped caudally in each zone. Thus there are four body maps in the PLLL. The medial segment receives ampullary input (Heiligenberg and Dye, '82) and maps the dorsoventral body axis mediolaterally, as does the tuberous centrolateral segment. The tuberous centromedial and lateral segments map the dorsoventral axis lateromedially. Thus the medial and centromedial segments meet belly to belly, the centromedial and centrolateral segments meet back to back, and the centrolateral and lateral segments meet belly to belly. Adjacent electrosensory maps within the PLLL are therefore always mirror images.     

Multiple electrosensory maps in the medulla of weakly electric gymnotiform fish. I. Physiological differences

The electrosensory lateral line lobe in the weakly electric gymnotiform fish Eigenmannia contains 3 topographic maps of high-frequency (tuberous) electroreceptive information from the body surface. The maps receive identical primary afferent input since axonal collaterals of both amplitude- and phase-coding afferents project to all 3 maps (Heiligenberg and Dye, 1982). Response properties of the amplitude-coding pyramidal neurons in the multiple maps were investigated in order to determine whether the maps differ physiologically. Units in the lateral map have larger receptive fields and are more sensitive than units in the centromedial map. The former units respond more phasically and with shorter latencies to step changes in stimulus amplitude (measured from the stimulus onset to the maximum response). Although 75% of pyramidal cells in all maps show a center-surround receptive-field organization, the strength of the inhibitory surround varies among maps, tending to be weakest for units in the lateral map and strongest for units in the centromedial map. Pyramidal neurons also differ in their responses with respect to the temporal frequency of amplitude modulations; the majority of units in the lateral map prefer high temporal frequencies, while those in the centromedial map prefer low frequencies. These results suggest that the multiple electrosensory maps could provide the initial separation of spatial and temporal processing of sensory information, much as has been suggested for X and Y ganglion cells in the cat retina (Shapley and Perry, 1986). The centromedial map could provide high spatial contrast with correspondingly poor temporal resolution, while the more sensitive units in the lateral map could best provide information about temporal changes in stimulus amplitude.     

Multiple electrosensory maps in the medulla of weakly electric gymnotiform fish. II. Anatomical differences

Both wave- and pulse-type species of weakly electric gymnotiform fish have 3 topographic maps of electroreceptive information in the electrosensory lateral line lobe (ELL). These maps receive identical input from trifurcating axons of phase- and amplitude-coding primary afferents (Carr et al., 1982; Heiligenberg and Dye, 1982). Physiological experiments in the ELL of the wave-type fish Eigenmannia show that the amplitude-coding pyramidal cells differ among maps with respect to receptive field size, sensitivity, rate of adaptation, and temporal-frequency response (Shumway, 1989). This study investigated morphological correlates of the physiological differences among maps. Estimates of primary afferent convergence in Eigenmannia, based on map size, cell counts, and areas of terminal fields from intracellularly filled P-type primary afferents, suggest a 2-fold increase in convergence in the lateral map relative to the centromedial map. Similar differences in convergence between maps are found in the wave-type species Apteronotus leptorhynchus and the pulse-type fish Hypopomus occidentalis. The lateral and centrolateral maps in Hypopomus, however, show an even greater difference in convergence. Comparison of the efferent projections of pyramidal cells among the different maps of Eigenmannia indicates that cells from the 3 maps terminate in the same laminae of the torus semicircularis, but the maps differ in the strength of projection to particular laminae. In both wave-type species, the abundance of a class of interneurons which receives descending input and inhibits pyramidal cells (interneurons of the ventral molecular layer) differs among maps; the centromedial map has 10 times fewer neurons of this type than the other 2 maps. Cytochrome oxidase studies in all 3 species demonstrated increased levels of activity in the lateral map, within the region receiving descending input from the cerebellum. These results suggest that the primary anatomical bases of the physiological differences among maps are differences in the amount of primary afferent convergence, coupled with differences in descending input.     

Correlating gamma-aminobutyric acidergic circuits and sensory function in the electrosensory lateral line lobe of a gymnotiform fish

Electric fish generate an electric field, which they sense with cutaneous electroreceptors. Electroreceptors project topographically onto the medullary electrosensory lateral line lobe (ELL). The ELL of gymnotiform electric fish is divided into four segments specialized to detect different aspects of the electrosensory input; it is also laminated with separate laminae devoted to electroreceptive input, interneurons, projection neurons, and feedback input. We have utilized antisera to glutamic acid decarboxylase (GAD) and γ-aminobutyric acid (GABA) to map the distribution of GABAergic cells and fibers in the ELL of the gymnotiform fish, Apteronotus leptorhynchus. Six types of GABAergic interneurons are found in ELL: Type 2 granular cells (granular layer) project to pyramidal cells; polymorphic cells (pyramidal cell layer) project to the non-GABAergic type 1 granular cells; ovoid cells (deep neuropil layer) project bilaterally upon basilar dendrites of pyramidal cells; multipolar cells (deep neuropil layer) project bilaterally, probably to dendrites and neurons within the deep neuropil layer; and neurons of the ventral molecular layer and stellate cells (molecular layer) project to apical dendrites of pyramidal cells. GABAergic bipolar cells in the nucleus praeminentialis, a rhombencephalic structure devoted to feedback in the electrosepsory system, project in relatively diffuse fashion to pyramidal cells. We hypothesize that the various GABAergic circuits of the ELL can be correlated with specific functions: type 2 granular cells with adaptation, size of receptive field center, and gain; polymorphic cells and type 1 granular cells with regulation of surround inhibition; ovoid cells with common mode rejection; and neurons of the ventral molecular layer with adaptive gain control. The feedback GABAergic input from bipolar cells of n. praeminentialis to pyramidal cells may be part of a searchlight mechanism similar to the one postulated for thalamocortical systems. © 1994, Wiley-Liss, Inc.     

Posterior lateral line afferent and efferent pathways within the central nervous system of the goldfish with special reference to the Mauthner cell

The goldfish posterior lateral line nerve consists of a dorsal and a ventral branch, each of which is associated with a ramus of the sensory branch of the VII th nerve (ramus recurrens facialis). The afferent and efferent pathways of these nerves within the central nervous system were studied by using horseradish peroxidase (HRP) histochemistry. The afferent fibers of the ramus recurrens facialis travel in the ventral portion of the VIIth nerve as it enters the brain and project predominantly to the ipsilateral half of the facial lobe. The afferent fibers of either the dorsal or ventral branch of the posterior lateral line nerve split into two bundles as they enter the brain. The caudally projectingfascicle terminates predominantly in thenucleusmedialis. The fibers of the rostrally projecting bundle terminate predominantly in nucleus medialis and nucleus magnocellularis and in the eminentia granularis. The posterior lateral line efferent somata were located in the diencephalon as well as in the medulla oblongata. The medullary efferent neurons formed two distinct groups, a rostral and a caudal nucleus. The cell bodies of the latter were more numerous and larger than those of the former. The axons of the efferent neurons exit from the brain by one of two routes. The first is at the level of the rostral efferent nucleus and the second at the level of the Mauthner cell. Previous reports have described input of posterior lateral line afferent fibers to the Mauthner cell soma and proximal lateral dendrite of the goldfish. This electrophysiological input was bilateral and was interpreted as monosynaptic. The afferent input described in this study was ipsilateral and ended in the vicinity of the distal lateral dendrite. These differences are discussed in the context of the neuronal circuitry that may be present.     

Capsaicin-induced neuronal degeneration: silver impregnation of cell bodies, axons, and terminals in the central nervous system of the adult rat

Capsaicin is a neurotoxic substance valued in neurobiological research because of its ability to selectively damage small unmyelinated primary sensory neurons. Previous work has indicated that systemic capsaicin administration causes permanent neuronal degeneration in neonatal rats, but evidence that capsaicin has a similar effect in adults is equivocal and incomplete. Therefore, we used silver impregnation, a method that labels degenerating neurons, to examine the central nervous system of adult rats after systemic capsaicin treatment. Adult rats were injected with a single intraperitoneal dose of capsaicin (50 or 90 mg/kg) or vehicle solution and killed 6, 12, 18, 24, 48, 96, or 240 hours later. Sections of brain and spinal cord were stained with the Carlsen-de Olmos cupric silver method. As reported previously, stained sections revealed degeneration in areas known to be innervated by small-diameter primary sensory fibers: the substantia gelatinosa of the spinal cord dorsal horn and spinal trigeminal nucleus, the solitary nucleus and tract, and the lateral borders of the area postrema. In addition, axon and terminal degeneration was observed in several discrete forebrain and hindbrain areas not previously associated with capsaicin-induced degeneration in either adult or neonatal rats: the inferior olive, the olivary pretectal nucleus, the interpeduncular nucleus, the suprachiasmatic nucleus, and the lateral septum/medial accumbens region. Furthermore, degenerating cell bodies were observed in the intrafascicular nucleus of the ventromedial midbrain tegmentum, in the supramammillary nucleus, and in the posterior hypothalamic area. Unilateral nodose ganglionectomy produced terminal staining on the denervated side very similar to that induced bilaterally by capsaicin. In addition, unilateral nodose ganglionectomy 1 month prior to capsaicin injection greatly attenuated staining in the ipsilateral nucleus of the solitary tract, confirming the hypothesis that capsaicin damages vagal sensory neurons innervating this nucleus. Capsaicin-induced damage in adult rats was long-lasting, since the second of two capsaicin treatments spaced 4.5 months apart produced no additional degeneration.     

Hormone-mediated plasticity in the electrosensory system of weakly electric fish

Weakly electric fish communicate with electric signals that are species-specific and often sexually dimorphic. These signals are produced from an electric organ in the tail that is under the control of a command nucleus in the medulla. The sexual dimorphism of the discharge is due to sex differences in the rhythmic firing of the medullary pacemaker nucleus and the duration of the electric organ pulse. These fish detect their own discharges and those of conspecifics by a class of sensory receptors called tuberous electroreceptors. The electroreceptors of each species are narrowly tuned to the frequencies of its electric organ discharge, and each sex is most sensitive to its own discharge frequency range. Sex steroid hormones have the capacity to influence and coordinate the electrical activity of the three target tissues that constitute the electrosensory system. This is proving to be a model system in which to study the action of steroids on the electrical activity of excitable cells.     

Larval electroreceptors in the epidermis of mormyrid fish: I. Tuberous organs of type A and B

Two types of larval electroreceptors, type A and B, are described in the epidermis of the head of larvae of three mormyrid species, Campylomormyrus cassaicus, Mormyrus rume proboscirostris and Pollimyrus isidori, bred in captivity. In each of these electroreceptor organs, a single sensory cell is found inside an intraepidermal cavity, sitting on a platform of accessory cells. The cavity is filled with microvilli originating both from the sensory cell and from the epidermal covering cells lining the intraepidermal cavity. These two types of tuberous larval electroreceptors differ in their distribution in the epidermis of the head, in the composition of their accessory cells, and by their innervation. The innervation found in type B organs is similar to that already described for electroreceptors of adult mormyrids. The sensorineural junction is composed of primary afferent terminal boutons, which contact the base of the sensory cell. Opposite each terminal bouton, a ribbon-like synaptic bar surrounded by vesicles is found in the cytoplasm of the sensory cell. In contrast, the base of the sensory cell in type A larval electroreceptors is not contacted by nervous terminal boutons, but instead forms closed appositions with specialized prolongations of accessory cells of the platform. The base of the sensory cell presents membrane evaginations, with hemispheric synaptic structures and few synaptic vesicles. These two types of electroreceptor organs degenerate at the time of the degeneration of the larval electric organ and the functional differentiation of the adult electric organ. The functional role of two tuberous electroreceptor types is examined.     

Larval electroreceptors in the epidermis of mormyrid fish: II. The promormyromast

Promormyromasts were found in the epidermis of the head of the larvae of five species of mormyrids bred in captivity. The promormyromast is a larval electroreceptor belonging to the specific lateral line system. In 12-day-old larvae this electroreceptor is characterized by a single sensory cell and two types of accessory cells. One type of accessory cell has dark cytoplasm, few microtubules, and contacts the sensory cell directly, whereas a second type has pale cytoplasm, many microtubules, and forms an outer layer not directly in contact with the sensory cell. This second type is referred to as a long pyriform accessory cell. This assembly of cells is situated below an intraepidermal cavity filled with acid polysaccharides. The bordering epidermal cells extend microvilli into the intraepidermal cavity. The apexes of the sensory cell, and of the two types of accessory cells, also open into the intraepidermal cavity but bear no microvilli. The promormyromast is innervated by an unmyelinated sensory nerve fiber passing through the basal membrane, which then splits into several branches between the accessory cells. These branches contact the periphery of the sensory cell with terminal boutons. At the site of each contact a ribbon-like structure surrounded by vesicles is present in the cytoplasm of the sensory cell. In older larvae of Campylomormyrus cassaicus, membrane foldings develop at the periphery of the pyriform accessory cells and accessory cell staining properties change just before transformation to become a mormyromast. The functional role of the promormyromast of the larval mormyrids is discussed.     

Light and electron microscopical studies on the spherical neurons in the electrosensory lateral line lobe of the gymnotiform fish,Sternopygus

Spherical cells are a principal cell type of the electrosensory lateral line lobe (ELLL) and play a crucial role in the jamming avoidance response (JAR) behavior. Since Sternopygus, a low frequency gymnotiform genus, does not display a JAR we searched for spherical cells in its ELLL. While present in Sternopygus, spherical cells differed remarkedly from those in the high-frequency gymnotiforms, Eigenmannia and Apteronotus. This study reveals species-characteristic differences in the morphology and synaptology of the spherical cell, a projection neuron located in the deep neuropil layer (DNL) of the ELLL. In contrast to the adendritic spherical cell of other species, the spherical neuron in Sternopygus exhibits an extensive basilar dendrite that extends into the primary electroreceptive afferent zone, the deep fiber layer (DFL). In Sternopygus, these neurons are distributed evenly across the full length of each tuberous subdivision, with cell densities highest in the centrolateral subdivision. At the ultrastructural level, the contacts on the soma, proximal, and distal dendrite of the spherical neuron in Sternopygus are asymmetrical chemical synapses, quite distinct from the electrotonic gap junctions found on the spherical neurons of other species.     

Decreased striatal dopamine D2 receptor binding in amyotrophic lateral sclerosis (ALS) and multiple system atrophy (MSA): D2 receptor down-regulation versus striatal cell degeneration

Recently, decreased striatal dopamine D2-receptor binding was demonstrated in vivo in amyotrophic lateral sclerosis (ALS). To further elucidate the pathogenetic mechanism underlying this D2-receptor deficit, a multi-level comparison was made between 30 sporadic ALS subjects and 24 patients with multiple system atrophy (MSA), a disorder clinically characterized by bradykinesia, neuroradiologically by severe D2-receptor loss, and neuropathologically by degenerating striatal cells. The extent of D2-deficit in ALS and MSA were within the same range, but extrapyramidal signs and symptoms were virtually absent in our ALS patients. Striatal cell loss in general or competitive D2-receptor occupancy could be considered unlikely in ALS. The striatum receives massive glutamatergic input and the pathogenesis of ALS may be related to increased glutamatergic excitotoxicity. As other mechanisms (cell loss, receptor occupancy) could be ruled out, and as animal studies suggest that (excess of) glutamate decreases striatal D2-receptor synthesis, the striatal D2-receptor deficit in ALS is most likely to be caused by a receptor down-regulation.     

Postembryonic development of the cerebellum in gymnotiform fish

In contrast to adult mammals, adult teleost fish regularly generate new neurons and glial cells in many brain regions. A previous quantitative mapping of the proliferation zones in the brain of adult Apteronotus leptorhynchus (Teleostei, Gymnotiformes) has shown that 75% of all mitotically active cells are situated in the cerebellum (Zupanc and Horschke [1995] J. Comp. Neurol. 353:213–233). By employing the thymidine analogue 5-bromo-2′-deoxyuridine, we have, in the present study, investigated the postembryonic development of this brain region in detail. In the corpus cerebelli and the valvula cerebelli, the vast majority of newborn cells originate in the respective molecular layers. Within the first few days of their life, these cells migrate toward specific target areas, namely, the respective granule cell layers. In the caudal part of the cerebellum, the granule cell layer of the eminentia granularis pars medialis displays the highest mitotic activity. From there, the cells migrate through the adjacent molecular layer to the granule cell layer of the eminentia granularis pars posterior. Combination of retrograde-tracing techniques with immunohistochemistry for 5-bromo-2′-deoxyuridine showed that at least a portion of the newly generated cells develop into granule neurons. Many of the newly generated cells survive for long periods of time. A large fraction of these cells is added to the population of already existing cells, thus resulting in a permanent growth of the target areas and their associated structures. © 1996 Wiley-Liss, Inc.     

Targeting cell surface receptors for axon regeneration in the central nervous system

Axon regeneration in the CNS is largely unsuccessful due to excess inhibitory extrinsic factors within lesion sites together with an intrinsic inability of neurons to regrow following injury. Recent work demonstrates that forced expression of certain neuronal transmembrane receptors can recapitulate neuronal growth resulting in successful growth within and through inhibitory lesion environments. More specifically, neuronal expression of integrin receptors such as alpha9beta1 integrin which binds the extracellular matrix glycoprotein tenascin-C, trk receptors such as trk B which binds the neurotrophic factor BDNF, and receptor PTPσ which binds chondroitin sulphate proteoglycans, have all been show to significantly enhance regeneration of injured axons. We discuss how reintroduction of these receptors in damaged neurons facilitates signalling from the internal environment of the cell with the external environment of the lesion milieu, effectively resulting in growth and repair following injury. In summary, we suggest an appropriate balance of intrinsic and extrinsic factors are required to obtain substantial axon regeneration.     

The laminar distribution of amino acids in the caudal cerebellum and electrosensory lateral line lobe of weakly electric fish (Gymnotidae)

We have studied the distribution of the putative amino acid neurotransmitters glutamate, aspartate, gamma-aminobutyric acid (GABA), glycine, taurine and beta-alanine in the caudal cerebellar lobe and electrosensory lateral line lobe (ELL) of weakly electric gymnotid fish. In the caudal lobe of the cerebellum, the levels of the various amino acids in the granular and molecular layers are comparable to the levels in the rat cerebellum, with the exception of taurine which is present in greater amounts in the gymnotid. In the ELL, these amino acids are differentially distributed in the various layers of this structure. Glutamate and taurine are enriched in the molecular layer, whereas GABA, aspartate, and beta-alanine are enriched in the deep neuropil + granular layers. Glycine is slightly enriched in the pyramidal cell layer.     

Immunohistochemical localization of ryanodine binding proteins in the central nervous system of gymnotiform fish.

The ryanodine receptor, an integral membrane protein of the sarcoplasmic reticulum in muscle, embodies a high conductance channel permeable to calcium ions. Recent studies have identified ryanodine-binding proteins in avian and mammalian central nervous systems. These neuronal ryanodine receptors appear to function as Ca2+ channels which may gate the release of Ca2+ from caffeine-sensitive intracellular pools in neurons. In the present investigation, we employed monoclonal antibodies against ryanodine-binding proteins of avian muscle cells to the brain of weakly electric gymnotiform fish. Immunoprecipitation and Western blot analysis revealed two isoforms in the fish brain, with molecular weights comparable to those of avian and fish muscle ryanodine-binding proteins. By employing immunohistochemical techniques, we mapped these proteins in fish brain. Ryanodine receptor-like immunoreactivity was found in nerve cell bodies as well as dendrites and axonal processes. The ryanodine-binding protein is distributed throughout the neuraxis in specific cell types of the gymnotiform brain. In the telencephalon, immunoreactive cells were found in the glomerular layer of the olfactory bulb, in the supracommissural subdivision of the ventral telencephalon, and in the intermediate rostral subdivision of the ventral telencephalon. In the diencephalon, immunoreactive cells or fibers were observed in the nucleus prethalamicus and the habenula, within the nucleus at the base of the optic tract and the adjacent dorsal tegmental nucleus, the pretectal nuclei A and B, and the nucleus electrosensorius. In addition, immunopositive cells were seen in several nuclei of the hypothalamus, with the inferior and lateral subdivision of the nucleus recessus lateralis displaying the highest concentration of neurons. In the mesencephalon, the optic tectum contained the greatest number of immunopositive cells. In the rhombencephalon, labelling was seen in the nucleus of the lateral valvula, central gray, lateral tegmental nucleus, in boundary cells of the nucleus praeminentialis, efferent octavolateral nucleus, an area adjacent to the medial edge of the lateral reticular nucleus, nucleus medialis, and electrosensory lateral line lobe. As in avian brain, cerebellar Purkinje cells were positive for ryanodine-binding protein, although only subsets of Purkinje cells were labelled.