GABA-like Immunoreactive Neurons Research Articles

The neuro-glandular brain of the Pyramicocephalus phocarum plerocercoid (Cestoda, Diphyllobothriidea): Immunocytochemical and ultrastructural study

А novel type of a complex neuro-glandular brain structure including both nervous and glandular elements and associated with sensory ones is detected in Pyramicocephalus phocarum plerocercoid (Cestoda: Diphyllobothriidea), parasite of Gadus morua from the White Sea. The brain has two lateral lobes connected by a long cellular median commissure. The brain is tightly surrounded by glandular cells, which receive numerous synapses from the brain neurons. A complex of sensory organs associated with ducts and terminal pores of the frontal glands lies in the scolex tegument. Serotonin, FMRFamide- and GABA-like immunoreactive (IR) neurons are found in the brain, the main nerve cords, and the plexus of the plerocercoid. The innervation of the frontal gland ducts by FMRFamide-IR neurites is detected for the first time proving that they function under control of the nervous system and thus evidencing the eccrine nature of the secretion mechanism. Ultrastructural data show that light, dark and neurosecretory neurons are present in the brain lobes. The median commissure consists of loosely arranged thin parallel axons and several giant and small neurons. The commissure is stratified and penetrated by frontal glandular cells and their processes. Such neuro-glandular morpho-functional brain complex is suggested as a model for Diphyllobothriidae family. Five structural types of sensory organs are described in the scolex of P. phocarum; their colocalization with eccrine gland terminals is supposedly specific for Diphyllobothriidae family. Within the order Diphyllobothriidea, there are significant differences in the architecture of the plerocercoid brain at the family level. We suppose homology of giant commissural neurons among Diphyllobothriidea. Differences between diphyllobothriidean nervous system and that of other cestodes are discussed.

GABA in the Nervous System of the CestodesDiphyllobothrium dendriticum(Diphyllobothriidea) andCaryophyllaeus laticeps(Caryophyllidea), with Comparative Analysis of Muscle Innervation

The nervous system (NS) of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea) was investigated using immunocytochemistry. The GABA neurotransmitter was identified in the NS of both species; GABAergic neurons were detected in the main nerve cords (MC). GABA-like immunoreactive neurons were predominantly unipolar and exhibited more intensive immunoreactivity in the neurite than in the perikaryon. In C. laticeps , GABA-like immunoreactive somas are located in both the MCs and peripheral NS near the longitudinal muscles. The innervation of the body musculature was studied using a combination of antibodies against GABA, serotonin (5-HT), and FMRFamide and with complementary staining of F-actin. In both species, the location of GABAergic neurites is associated with all muscle layers including the subtegumental, longitudinal, transverse, and dorsoventral muscles. The cytomorphology of 5-HTergic motoneurons in the MCs of both species is described and differences in muscle innervation between D. dendriticum and C. laticeps are demonstrated. No evidence for co-localization of GABA with 5-HT or FMRFamide neurotransmitters at any particular neuron was found. Neuropiles in MCs and peripheral NS had separate sets of immunoreactive neurites. A functional role for GABA in muscle innervation is discussed.

Changes in GABA and GABAB receptor expressions are involved in neuropathy in the rat cuneate nucleus following median nerve transection

This study examined the relationship between changes in GABA transmission and behavioral abnormalities after median nerve transection. Following unilateral median nerve transection, the percentage of GABA-like immunoreactive neurons in the cuneate nucleus and that of GABA(B) receptor-like immunoreactive neurons in the dorsal root ganglion in the injured side decreased and reached a nadir at 4 weeks after median nerve transection. Four weeks after bilateral median nerve transection and intraperitoneal application with saline, baclofen (2 mg kg⁻¹), or phaclofen (2 mg kg⁻¹) before unilateral electrical stimulation of the injured median nerve, we investigated the level of neuropeptide Y release and c-Fos expression in the stimulated side of the cuneate nucleus. The neuropeptide Y release level and the number of c-Fos-like immunoreactive neurons in the baclofen group were significantly attenuated, whereas those in the phaclofen group had increased compared to the saline group. These findings indicate that median nerve transection reduces GABA transmission, promoting injury-induced neuropeptide Y release and consequently evoking c-Fos expression in cuneate nucleus neurons. Furthermore, this study used the CatWalk method to assess behavioral abnormalities in rats following median nerve transection. These abnormalities were reversed by baclofen treatment. Overall, the results suggest that baclofen treatment block neuropeptide Y release, subsequently lessening c-Fos expression in cuneate neurons and consequently attenuating neuropathic signal transmission to the thalamus.

Sexual Dimorphism in Shape and Distribution of GABA-Like Immuno-Reactive Neurons in Cricket Terminal Abdominal Ganglion

We studied age-related sexual dimorphism of the terminal abdominal ganglion in the cricket, Gryllus bimaculatus using osmium/ethyl-gallate staining and GABA immunohistochemistry. We determinedthat the volume of the terminal abdominal ganglion increases with increasing body size. The growth ratio between longitude and width in the terminal abdominal ganglion is the same in all nymph stages and sexes. However, the longitudinal growth of the male terminal abdominal ganglion ceases in the 8th instar and the adult stage. Therefore, in the adult stage, sexual dimorphism is observed in ganglion morphology. Additionally, sexual dimorphism is also observed in numbers of neuronal somata and GABA-like immunoreactive neurons in the terminal abdominal ganglion. At the beginning of the 8th instar, the number of somata is greater in female crickets than in males. GABA-like immunoreactive neurons, which were classified into eight clusters, increased between the 8th instar and adult in both sexes, but their numbers were sexually dimorphic in the 7th and 8th instars, and in the adult. Females had a greater number of GABA-like immunoreactive neurons than males. These differences occurred mainly in the 8th neuromere cluster, as well as the anterior region of the 9-11th neuromeres.

Changes of μ-opioid receptors and GABA in visual cortex of chronic morphine treated rats

Electrophysiological and biochemical studies have indicated that GABAergic modulation is involved in the opioid-induced altering of response properties of visual cortical cells and impairing of short-term synaptic plasticity in the geniculo-cortical visual pathway. The aim of the current study was to examine whether there were changes in the localization and density of micro-opioid receptor subtype (MOR1) and GABA in the visual cortex of morphine-dependent and abstinent rats. Immunofluorescence histochemical method was applied to display MOR1 and GABA distribution. We found that MOR1-like immunoreactive neurons were significantly lowered in layer I-VI of visual cortex of rats sacrificed immediately after the last injection (defined as morphine-dependent (MD)) than saline-control group. In rats sacrificed just before the last injection (defined as morphine-abstinent (MA)), the density of micro-opioid receptor was higher than that in dependent group in layer I-V neurons of visual cortex, but remained lower than those in control group. Three hours after the last morphine injection (defined as 3h after morphine-abstinent (3h)), MOR1-like immunoreactive neurons in layer I and layer IV of visual cortex were still significantly lower than control. As to GABA-like immunoreactive neurons, they were significantly decreased in abstinent group compared to dependent group. These results provide morphological evidence that opioid-induced altering of response properties of visual cortical cells might be due to the change of visual cortical GABAergic system induced by opioid via mu-opioid receptor, which result in disinhibition of visual cortex projection neurons and their abnormal firing.

GABAergic neurons express μ-opioid receptors in the ventrolateral orbital cortex of the rat

Behavioral studies have indicated that GABAergic modulation is involved in the opioid-induced antinociception in the ventrolateral orbital cortex (VLO). The aim of the current study was to examine whether the GABAergic neurons in the rat VLO expressed μ-opioid receptor subtype 1 (MOR1). This study employed immunofluorescence histochemical double-staining technique and showed that a considerable amount of GABA- and MOR1-like immunoreactive neurons existed in layers II–VI in the VLO. Of these GABA-like immunoreactive neurons, 92.0% of them showed MOR1-like immunoreactivities. Similarly, 80.2% of MOR1-like immuoreactive neurons also exhibited GABA-like immunoreactivities. These results provide morphological evidence that opioid-induced antinociception in the VLO might be due to an inhibitory effect by opioid via MOR1 on GABAergic neurons, resulting in disinhibition of VLO projection neurons and leading to activation of the VLO-PAG brainstem descending pain control system to depress the nociceptive inputs at the spinal cord level.

Somatostatin-, calcitonin gene-related peptide, and γ-aminobutyric acid-like immunoreactivitity in the frog lumbosacral spinal cord: distribution and effects of sciatic nerve transection

Using immunohistochemistry and optical densitometry, somatostatin (SOM), calcitonin gene-related peptide (CGRP), and γ-aminobutyric acid (GABA) were investigated in the lumbosacral spinal cord of the frog Rana catesbeiana after sciatic nerve transection. In control animals, the densest network of the SOM-, CGRP- and GABA-like immunoreactive fibers was located in the dorsal part of the lateral funiculus. SOM and GABA-like fibers were found in the dorsal terminal field and in the mediolateral band. The latter region showed CGRP and SOM-like immunoreactive cell bodies. SOM- and GABA-like immunoreactive neurons also occurred around the cavity of the central canal, and other GABA-like fibers were found in the ventral terminal field. While the ventral horn showed scarce somatostatin-like fibers, the putative motoneurons were immunoreactive for the two peptides investigated and GABA, but only a few SOM- and GABA-like fibers occurred in the ventral funiculus. After axotomy, GABA-like immunoreactivity decreased in the dorsal part of the lateral funiculus on the same side of the lesion. The other regions remained labeled. These changes were observed at 3 days following axonal injury and persisted at 5, 8 and 15 days. There was no significant difference in the pattern of CGRP- and SOM- immunoreactivity between the axotomized and the control sides. These results are discussed in relation to the effects of the peripheral axotomy on GABA, SOM, and CGRP expression in vertebrates, emphasizing the use of frogs as a model to study the effects of peripheral nerve injury.

Ontogenesis of the snail, Helix aspersa: embryogenesis timetable and ontogenesis of GABA-like immunoreactive neurons in the central nervous system.

Late stages of embryogenesis in the terrestrial snail Helix aspersa L. were studied and a developmental timetable was produced. The distribution of gamma-aminobutyric acid-like immunoreactive (GABA-ir) elements in the CNS of the snail was studied from embryos to adulthood in wholemounts. In adults, approximately 226 GABA-ir neurons were located in the buccal, cerebral and pedal ganglia. The population of GABA-ir cells included four pairs of buccal neurons, three neuronal clusters in the pedal ganglia, two clusters and six single neurons in the cerebral ganglia. GABA-ir fibers were observed in all ganglia and in some nerves. The first detected pair of GABA-ir cells in the embryos appeared in the buccal ganglia at about 63-64% of embryonic development. Five pairs of GABA-ir cell bodies were observed in the cerebral ganglia at about 64-65% of development. During the following 30% of development three more pairs of GABA-ir neurons were detected in the buccal ganglia and over fifteen cells were detected in each cerebral ganglion. At the stage of 70% of development, the first pair of GABA-ir neurons was found in the pedal ganglia. In the suboesophageal ganglion complex, GABA-ir fibers were first detected at about 90% of embryonic development. In the posthatching period, the quantity of GABA-ir neurons reached the adult status in four days in the cerebral ganglia, and in three weeks in the pedal ganglia. In juveniles, transient expression of GABA was found in the pedal ganglia (fourth cluster).

Distribution and developmental changes in GABA-like immunoreactive neurons in the central nervous system of pond snail,Lymnaea stagnalis

We examined three-dimensionally the arrangement of gamma-aminobutyric acid (GABA)-like immunoreactive neurons in the central nervous system (CNS) of the pond snail, Lymnaea stagnalis, by a combination of immunohistochemistry and confocal laser scanning microscopy on whole-mount preparations. GABA-like immunoreactivity was detected in all ganglia of the adult CNS. The following distribution of immunoreactive cell bodies was noted in the adult snail. Buccal ganglia: one cell body and five pairs of cell bodies, cerebral ganglia: one pair of cell bodies, pedal ganglia: two single cell bodies, two pairs of cell bodies, and three pairs of cell clusters, and pleural ganglia: one pair of cell bodies. In the asymmetrical parietal ganglia, three cell bodies were located in the left parietal ganglion; three cell bodies and three cell clusters were located in the right parietal ganglion. In the single visceral ganglion, a few scattered individual cell bodies and a cell cluster were GABA-like immunoreactive. Our results showed that the occurrence of GABA is widely spread in the CNS of adult L. stagnalis. GABA-like immunoreactivity in the CNS was not detected in the embryo but was observed after hatching, although the number of stained cells was less than in the adult, with the exception of those in the cerebral ganglia where their number decreased with maturation. Our results provide detailed maps of the central GABA-like immunoreactive neurons in juveniles, immatures, and adults of L. stagnalis.

Distribution and developmental changes in GABA-like immunoreactive neurons in the central nervous system of pond snail, Lymnaea stagnalis

Distribution and developmental changes in GABA-like immunoreactive neurons in the central nervous system of pond snail, Lymnaea stagnalis

Distribution of GABA-like immunoreactive neurons and fibers in the central visual nuclei and retina of frog, Rana pipiens.

Immunocytochemistry was used to study the distribution of gamma-aminobutyric acid (GABA) throughout the central visual nuclei and retina in Rana pipiens. In the retina, GABA immunoreactivity (both somata and fibers) was observed in all layers except the outer nuclear layer (ONL). Contrary to earlier reports, about 30% of total neurons within ganglion cell layer (GCL) expressed GABA immunoreactivity. Double-labeling studies indicated that about half of the GABA-containing perikarya in the GCL were retinal ganglion cells (RGCs). In the diencephalon, intensely labeled GABA-immunoreactive neurons and nerve fibers were observed within the neuropil of Bellonci (nB) and corpus geniculatum (CG), while only immunoreactive puncta were found in the rostral visual nucleus (RVN). In the pretectal region, the posterior thalamic nucleus (nPT) contained the most intensely labeled GABA immunoreactive perikarya and nerve fibers in the entire brain. Lightly immunoreactive perikarya were also found in the large-celled nucleus lentiformis mesencephali (nLM), as well as in the pretectal gray, which contains neurons postsynaptic to the retinal terminal zones within nLM. In the optic tectum (OT), both immunoreactive perikarya and fibers were found within superficial layers 8 and 9, whereas only densely packed immunoreactive perikarya were evident in the deep tectal layers (i.e. 2, 4, 6). The nucleus of the basal optic root (nBOR) contained a small number of lightly labeled GABA-immunoreactive perikarya, mostly located in the dorsal half of the nucleus. A large number of perikarya within the nucleus isthmi (NI) were also immunostained.

Distribution of GABA-like immunoreactive neurons in insects suggests lineage homology.

Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in vertebrates and invertebrates (Sattelle [1990] Adv. Insect Physiol. 22:1-113). The GABA phenotype is lineally determined in postembryonic neurons in the tobacco hawkmoth, Manduca sexta (Witten and Truman, [1991] J. Neurosci. 11:1980-1989) and is restricted to six identifiable postembryonic lineages in the moth's thoracic hemiganglia. We used a comparative approach to determine whether this distinct clustering of GABAergic neurons is conserved in Insecta. In the nine orders of insects surveyed (Thysanura, Odonata, Orthoptera, Isoptera, Hemiptera, Coleoptera, Diptera, Lepidoptera, and Hymenoptera), GABA-like immunoreactive neurons within a thoracic hemiganglion were clustered into six distinct groups that occupied positions similar to the six postembryonic lineages in Manduca. On the basis of cell body position and axon trajectories, we suggest that these are indeed homologous lineage groups and that the lineal origins of the GABAergic cells have been very conservative through insect evolution. The distinctive clustering of GABA-positive cells is shared with crustaceans (Mulloney and Hall [1990] J. Comp. Neurol. 291:383-394; Homberg et al. [1993] Cell Tissue Res. 271:279-288) but is not found in the centipede Lithobius forficulatus. There is a two- to threefold increase in numbers of thoracic neurons between the flightless Thysanura and the most advanced orders of insects. Using the GABA clusters as indicators of specific lineages, we find that only selected lineages have significantly contributed to this increase in neuronal numbers.

The ontogeny of GABA- and glutamate-like immunoreactivity in the embryonic Australian freshwater crayfish, Cherax destructor

The distribution and ontogeny of GABA- and glutamate-like immunoreactivity in embryos of the Australian freshwater crayfish Cherax destructor were investigated over the period from 30% development until hatching. GABA-like immunoreactive cells and fibres appeared first in the brain at 40–45% development. By 70% development, GABA-like immunoreactive cells were present in almost all ganglia, and GABA-like immunoreactive fibres were distributed extensively throughout the neuropil, commissures and connectives of the central nervous system, and were also found in peripheral nerve roots supplying the appendages and the abdominal musculature. In contrast, glutamate-like immunoreactivity did not appear in the central nervous system until 60–65% development. By the time of hatching, the distribution of glutamate-like immunoreactivity was restricted to discrete regions of neuropil and fibre staining in the thoracic and abdominal nerve cord, the abdominal musculature and the appendages. The precocious establishment of the extensive distribution of GABA-like immunoreactive neurons in the developing crayfish embryo is consistent with the possibility that these neurons play a trophic role in controlling or modulating the development of the nervous system.

Three classes of GABA-like immunoreactive neurons in the mushroom body of the cockroach

The mushroom body (MB) is a higher center of the insect brain and is critical to some forms of associative memory. Each MB consists of calyces connected to α and β lobes via pedunculus. In the calyces, input neurons make synaptic connections with intrinsic neurons. In the pedunculus and lobes, intrinsic neurons make synaptic connections with output neurons. Here, the distribution of γ-aminobutyric acid (GABA)-like immunoreactivity in the MB of the cockroach Periplaneta americana was investigated, using an antiserum against a GABA–protein conjugate, to elucidate inhibitory pathways of the MB. We report that three classes of extrinsic neurons of the MB exhibit GABA-like immunoreactivity. The first is four large neurons which arborize in a diffuse neuropil surrounding the α lobe and project into whole areas of the calyces. Their cell bodies are 30–50 μm in diameter, among the largest in the brain. The second group is 7–9 neurons ascending from the circumesophageal connective and projecting into the calyces, which probably represent inhibitory input neurons. The third group is ca. 40 neurons with dendritic arborizations in the junction between the pedunculus and the lobes, which probably represent inhibitory output neurons.

Electron microscopic study of GABAergic synaptic innervation of neurotensin-immunoreactive neurons in the dorsal raphe nucleus

A double immunocytochemical method combining the preembedding avidin-biotin-peroxidase-complex technique and the postembedding immunogold technique was used to examine synaptic interactions between GABAergic and neurotensin-containing neurons in the same tissue sections of the dorsal raphe nucleus of the rat. Whereas the neurotensin-like immunoreactive perikarya rarely received synapses from GABA-like immunostaining axon terminals, the neurotensin-like immunoreactive dendrites frequently received synapses from GABA-like immunoreactive neurons. These results suggest that GABAergic neurons could modulate neurotensinergic neurons in the dorsal raphe nucleus through synaptic relations. The immunocytochemically identified local synaptic circuit in the dorsal raphe was discussed.

Electron microscopic study of GABAergic synaptic innervation of neurotensin-immunoreactive neurons in the dorsal raphe nucleus

A double immunocytochemical method combining the preembedding avidin-biotin-peroxidase-complex technique and the postembedding immunogold technique was used to examine synaptic interactions between GABAergic and neurotensin-containing neurons in the same tissue sections of the dorsal raphe nucleus of the rat. Whereas the neurotensin-like immunoreactive perikarya rarely received synapses from GABA-like immunostaining axon terminals, the neurotensin-like immunoreactive dendrites frequently received synapses from GABA-like immunoreactive neurons. These results suggest that GABAergic neurons could modulate neurotensinergic neurons in the dorsal raphe nucleus through synaptic relations. The immunocytochemically identified local synaptic circuit in the dorsal raphe was discussed.

Chemically defined neuron groups and their subpopulations in the glomerular layer of the rat main olfactory bulb

Chemically-defined neuron groups and their subpopulations in the glomerular layer of the rat main olfactory bulb were revealed immunocytochemically using antibodies agianst γ-amino butyric acid (GABA), tyrosine hydroxylase (TH), methionin-enkephalin-Arg 6 -Gly 7 - Leu 8 (ENK), calretinin (CR), calbindin-D28K (calbindin) and thyrotropin-releasing hormone (TRH). GABA-like immunoreactive (GABA-LIR) neurons and CR immunoreactive (CR-IR) neurons were most numerous; they were about 1.5–3 times more numerous than calbindin immunoreactive (calbindin-IR), TH immunoreactive (TH-IR), ENK-llike immunoreactive (ENK-LIR) and TRH-like immunoreactive (TRH-LIR) neurons. We identified at least three distinct chemically-defined neuron groups, GABA-LIR neurons, CR containing neurons and calbindin containing neurons, since these three neuron groups were almost separate from one another. On the other hand, TH-IR and ENK-LIR neurons were nearly included in and thus considered to be subpopulations of GABA-LIR and CR-IR neurons, respectively, for about 80% of these two neuron groups contained GABA-L and CR immunoreactivities, respectively. TRH-LIR neurons appeared to be divided into two subpopulations, one containing the GABA-L immunoreactivity and the other containing the CR immunoreactivity. Thus in the glomerular layer of the rat olfactory bulb, GABA-LIR, CR-IR and calbindin-IR cells could be considered to the three distinct chemically-defined neuron groups, whereas TH-IR, TRH-LIR and ENK-LIR neurons were regarded as their subpopulations. Furthermore, some neurons are supposed to contain three substances (e.g. GABA + TH + TRH, GABA + TRH + ENK, CR + TRH + ENK, GABA + TRH + CR) or a few might even contain four substances (e.g. GABA + TRH + CR + ENK). Preliminary quantitative analysis using the optical disector method showed percentages of these three main groups to total cells in the glomerular layer; that is, neuron groups containing GABA, CR and calbinsin were about 20%, 20% and 10%, respectively.

Chemically defined neuron groups and their subpopulations in the glomerular layer of the rat main olfactory bulb

Chemically-defined neuron groups and their subpopulations in the glomerular layer of the rat main olfactory bulb were revealed immunocytochemically using antibodies agianst γ-amino butyric acid (GABA), tyrosine hydroxylase (TH), methionin-enkephalin-Arg 6-Gly 7- Leu 8 (ENK), calretinin (CR), calbindin-D28K (calbindin) and thyrotropin-releasing hormone (TRH). GABA-like immunoreactive (GABA-LIR) neurons and CR immunoreactive (CR-IR) neurons were most numerous; they were about 1.5–3 times more numerous than calbindin immunoreactive (calbindin-IR), TH immunoreactive (TH-IR), ENK-llike immunoreactive (ENK-LIR) and TRH-like immunoreactive (TRH-LIR) neurons. We identified at least three distinct chemically-defined neuron groups, GABA-LIR neurons, CR containing neurons and calbindin containing neurons, since these three neuron groups were almost separate from one another. On the other hand, TH-IR and ENK-LIR neurons were nearly included in and thus considered to be subpopulations of GABA-LIR and CR-IR neurons, respectively, for about 80% of these two neuron groups contained GABA-L and CR immunoreactivities, respectively. TRH-LIR neurons appeared to be divided into two subpopulations, one containing the GABA-L immunoreactivity and the other containing the CR immunoreactivity. Thus in the glomerular layer of the rat olfactory bulb, GABA-LIR, CR-IR and calbindin-IR cells could be considered to the three distinct chemically-defined neuron groups, whereas TH-IR, TRH-LIR and ENK-LIR neurons were regarded as their subpopulations. Furthermore, some neurons are supposed to contain three substances (e.g. GABA + TH + TRH, GABA + TRH + ENK, CR + TRH + ENK, GABA + TRH + CR) or a few might even contain four substances (e.g. GABA + TRH + CR + ENK). Preliminary quantitative analysis using the optical disector method showed percentages of these three main groups to total cells in the glomerular layer; that is, neuron groups containing GABA, CR and calbinsin were about 20%, 20% and 10%, respectively.

Immunoelectron microscopy of enkephalinergic innervation of GABAergic neurons in the periaqueductal gray

The pre-embedding double immunoreaction method was used to study synaptic relations of enkephalinergic and GABAergic neuronal elements in the ventrolateral part of the periaqueductal gray of the Wistar albino rat. The enkephalin-like neuronal elements were immunoreacted by the silver-gold intensified peroxidase-antiperoxidase method and the GABA-like immunoreactive neurons were immunoreacted by the unintensified peroxidase-antiperoxidase method. GABA-like immunoreactive neuronal somata were post-synaptic to both the enkephalin-like immunoreactive and the non-immunoreactive axon terminals. Enkephalin-like immunoreactive axon terminals were found to make synapses with GABA-like immunoreactive and non-immunoreactive dendrites. The synapses between the two kinds of chemically characterized neurons appeared to be both asymmetrical and symmetrical. Possible functional activity related to pain modulation, and synaptic relations between the enkephalinergic and GABAergic neurons in the periaqueductal gray and the dorsal raphe nucleus, are discussed.

Distribution and anatomy of GABA-like immunoreactive neurons in the central and peripheral nervous system of the snail Helix pomatia.

Gamma-aminobutyric acid (GABA)-like immunoreactive neurons were studied in the central and peripheral nervous system of Helix pomatia by applying immunocytochemistry on whole-mount preparations and serial paraffin sections. GABA-immunoreactive cell bodies were found in the buccal, cerebral and pedal ganglia, but only GABA-immunoreactive fibers were found in the viscero-parietal-pleural ganglion complex. The majority of GABA-immunoreactive cell bodies were located in the pedal ganglia but a few could be found in the buccal ganglia. Varicose GABA-ir fibers could be seen in the neuropil areas and in distinct areas of the cell body layer of the ganglia. The majority of GABA-ir axonal processes run into the connectives and commissures of the ganglia, indicating an important central integrative role of GABA-immunoreactive neurons. GABA may also have a peripheral role, since GABA-immunoreactive fibers could be demonstrated in peripheral nerves and the lips. Glutamate injection did not change the number or distribution of GABA-immunoreactive neurons, but induced GABA immunoreactivity in elements of the connective tissue ensheathing the muscle cells and fibers of the buccal musculature. This shows that GABA may be present in different non-neural tissues as a product of general metabolic pathways.