Glutamic Acid Decarboxylase-like Immunoreactivity Research Articles

Electrophysiological properties of cultured neonatal rat dorsal horn neurons containing GABA and met-enkephalin-like immunoreactivity.

We have developed a culture of neurons dissociated from the most superficial laminae of the neonatal rat spinal cord dorsal horn (DH). By using the perforated patch-clamp technique, we distinguished four types of neurons based on their firing properties in response to intracellular injection of 900 ms lasting current pulses. Type 1 neurons were characterized by a tonic firing. Type 2 neurons displayed marked spike accommodation and fired brief (<500 ms) bursts of action potentials, whereas type 3 neurons fired a single spike. Type 4 neurons exhibited different types of firing patterns, but all of them possessed a time-dependent inwardly rectifying current activated by membrane hyperpolarization. Met-enkephalin-like immunoreactivity (met-ENK-LI) and glutamic acid decarboxylase-like immunoreactivity (GAD-LI) were colocalized in 42% of the neurons (n = 59), which were previously identified electrophysiologically. Type 1-4 neurons represented respectively 4, 64, 20, and 12% of the population of neurons colocalizing met-ENK-LI and GAD-LI. We conclude that the electrophysiological properties of DH neurons present in our cultures are similar to those described in acute slice or hemisected spinal cord preparations and that met-ENK-LI and GABA-LI are preferentially colocalized in type 2 neurons.

Effects of excitatory amino acids on immunocytochemically identified populations of neurons in turtle retina

Excitatory amino acids play an important role in visual processing in the retinas of many species, but little is known about the identity of the specific postsynaptic cell types and the pharmacology of their receptors. To investigate which specific cell types were affected by excitatory amino acids, we examined the effects of exogenous aspartate, glutamate, kainic acid, N-methyl-D-aspartate, and MK-801 on retinal neurons. Specific populations of neurons were labelled using antibodies directed against glucagon, enkephalin, neurotensin, gamma-aminobutyric acid, glutamic acid decarboxylase, serotonin, glycine, glutamate or aspartate. We analyzed a combination of long-term in vivo injections (seven days following an intraocular injection of kainic acid) and short term in vitro incubations. There were changes in the labelling intensity and sometimes in the relative localization of all of the antigens in the drug treated retinas. Some observations suggested that the drugs were altering neurotransmitter metabolism. Differential responses were seen in specific cell types within the populations of neurons with neurotensin-, glutamate-, aspartate-, glycine, gamma-aminobutyric acid-, and glutamic acid decarboxylase-like immunoreactivity. The immunocytochemical approach used in these studies was able to determine specific retinal cell types which were influenced by particular excitatory amino acids. The broad extent of cell types influenced and the potential metabolic effects suggest that excitatory amino acids and their receptors play a complex role in visual processing.

The distribution of neurotransmitters and neurotransmitter‐related enzymes in the dorsomedial telencephalon of the pigeon (Columba livia)

Immunoreactivity to four neurotransmitters/transmitter-related enzymes was found in the dorsomedial telencephalon (hippocampal region) of the pigeon. Putative afferent fibers containing choline acetyltransferase-like, serotonin-like, and tyrosine hydroxylase-like immunoreactivity were seen in a fiber tract passing through the septo-hippocampal junction and along the medial wall of the hippocampal region. The most intensive labeling of neuropil and terminals of all four substances was found in the dorsomedial area of the hippocampal region. Glutamic acid decarboxylase-like immunoreactivity was seen in sparsely scattered cells throughout the region. These results are discussed in relation to hypotheses about the boundaries and subdivisions of the hippocampal region of the pigeon.

Peripheral inflammation is associated with increased glutamic acid decarboxylase immunoreactivity in the rat spinal cord

We have examined the frequency and distribution of neuron profiles immunoreactive for glutamic acid decarboxylase, a biosynthetic enzyme for the putative inhibitory neurotransmitter, gamma aminobutyric acid, in the lumbar spinal cord of colchicine-treated rats with unilateral inflammation of a hindpaw. Ipsilateral to the inflamed hindpaw, there was an apparent increase in the levels of glutamic acid decarboxylase, as indicated by significant increases in the number of visible glutamic acid decarboxylase-like immunoreactive profiles within the superficial dorsal horn, neck of the dorsal horn and the deep gray matter at L4. An increase limited to the deep gray matter at L6 was also seen. No alteration was identified at L2. These results are the first to demonstrate that peripheral inflammation is associated with altered levels of glutamic acid decarboxylase-like immunoreactivity.

The coexistence of substance P- and glutamic acid decarboxylase-like immunoreactivity in entopeduncular neurons of the rat

The coexistence of substance P- and glutamic acid decarboxylase-like immunoreactivity in cell bodies was investigated in the entopeduncular neurons of the rat by the double-immunofluorescence method using species-specific antibodies. Over half of the substance P-like immunoreactive cells were GABAergic. Double-labeled neurons were generally seen in the rostral to the middle region of the entopeduncular nucleus. Since the rostral portion of the entopeduncular nucleus mainly projects to the lateral habenular nucleus, it is suggested that double-labeled neurons in the entopeduncular nucleus largely project to the lateral habenular nucleus which is involved in the limbic system.

Vestibulospinal pathway in rabbit includes GABA-synthesizing neurons

When Fast blue is injected into the rabbit spinal cord it is retrogradely transported into nerve cell bodies located in the medial and the descending vestibular nuclei. Approximately 50% of the Fast blue-positive cells also contain glutamic acid decarboxylase-like immunoreactivity. These neurons presumably synthesize γ-aminobutyric acid (GABA) and the rabbit vestibulospinal pathway therefore contains a substantial inhibitory component.

Glutamic acid decarboxylase-like immunoreactivity in brainstem auditory nuclei of the rat.

The distribution of GABA-producing neurons in the brainstem auditory nuclei of the rat was investigated immunohistochemically by using an antibody to glutamic acid decarboxylase (GAD). In the cochlear nuclei, GAD immunoreactive neurons are present only in the superficial granular and molecular layers, whereas terminals are found in all subdivisions of the nuclei and are particularly dense surrounding large spherical cells and one type of stellate cell. In the superior olivary complex, GAD immunoreactive neurons are located in the lateral olivary nucleus and throughout the periolivary region. Immunoreactive terminals are distributed along dendrites of principal cells of the medial and lateral olivary nuclei and are clustered around somata of globular neurons of the nucleus of the trapezoid body. An extremely dense band of immunoreactive somata and terminals is present along the ventral edge of the olivary complex. The ventral, intermediate, and dorsal nuclei of the lateral lemniscus contain small fusiform GAD-immunoreactive neurons and a moderately dense plexus of immunoreactive terminals. The inferior colliculus contains a large population of GAD-immunoreactive perikarya and an extremely dense accumulation of immunoreactive terminals in the central, dorsomedial, and external nuclei. These observations indicate that GABA systems are involved in function at all levels of the brainstem auditory pathway.

The time of origin of cells demonstrating glutamic acid decarboxylase-like immunoreactivity in the hippocampal formation of the rat

Using the combination of [ 3H]thymidine autoradiography and immunohistochemistry for the detection of glutamic acid decarboxylase (GAD), we have determined that most of the cells demonstrating GAD-like immunoreactivity in the hippocampal formation of the rat become postmitotic on fetal day 14, though some cease dividing on fetal days 13 and 15–18. GAD-positive cells in all fields of the hippocampal formation appear to differentiate during roughly the same period though there is evidence for both septotemporal and regional variations in the pattern of development.

Horizontal cells of the mouse retina contain glutamic acid decarboxylase-like immunoreactivity during early developmental stages

We have used an antiserum to L-glutamic acid decarboxylase ((GAD), a synthesizing enzyme for gamma-aminobutyric acid (GABA)) to localize putative GABAergic neurons in the developing C57BL/6J mouse retina. At early developmental stages (embryonic day 17 to postnatal day 3), strong GAD-like immunoreactivity is detectable in cell bodies located within the neuroblastic layer. These cells have relatively large cell bodies and extend several sturdy processes which are oriented radially at these early stages. We have identified these cells as horizontal cells. In addition, cell bodies adjacent to the inner plexiform layer and both diffuse and punctate structures within the inner plexiform layer proper have weak GAD-like immunoreactivity at this time. By postnatal day 6, GAD-positive horizontal cell processes begin to form a horizontal network in the newly formed outer plexiform layer. Immunolabeling of amacrine cell bodies and of punctate structures in the inner plexiform layer becomes much stronger at this time, reaching a maximum staining intensity during the second postnatal week. After postnatal day 12, GAD-like immunoreactivity of the horizontal cells begins to decline; in 4-week-old mice the horizontal cells are no longer detectably labeled by this GAD antiserum. At the same time, the GAD-like-immunoreactive material in the inner plexiform layer becomes stratified, forming distinct layers. Amacrine cells and the inner plexiform layer remain GAD positive into adulthood.

Interpeduncular nucleus organization in the rat: Cytoarchitecture and histochemical analysis

The organization of the interpeduncular nucleus (IPN) in the adult rat was analyzed using cytoarchitectonic, histochemical and immunohistochemical methods. Four paired and four unpaired subnuclei can be distinguished in the IPN on the basis of neuronal size, morphology, staining characteristics and packing density. The rostral portion of the IPN contains a rostral dorsal, a rostral ventral and paired rostral lateral and dorsal lateral nuclei. The dorsal lateral nuclei continue into the caudal IPN, which also contains a caudal dorsal, a caudal ventral and paired caudal lateral nuclei. The distribution of extrinsic afferents and of chemically identified intrinsic neuronal and fiber populations within subdivisions of the IPN was examined using immunohistochemistry, acetylcholinesterase histochemistry, catecholamine histofluorescence and the autoradiographic tracing method. Six immunohistochemically distinct neuronal groups are identified in the IPN. Perikarya and axons showing substance P-, leu-enkephalin-, somatostatin-, avian pancreatic polypeptide-, serotonin- and glutamic acid decarboxylase-like immunoreactivity are localized to specific IPN subnuclei. Acetylcholinesterase-positive staining, extrinsic norepinephrine-containing fibers and afferents from the dorsal tegmental nuclei are also distributed specifically to IPN subnuclei. These findings demonstrate a cytoarchitectonic and cytochemical complexity in the rat IPN that implies an important functional role for this poorly understood nuclear complex.