REGIONAL DISTRIBUTION OF GLUTAMATE DECARBOXYLASE AND GABA WITHIN THE AMYGDALOID COMPLEX AND STRIA TERMINALIS SYSTEM OF THE RAT

The distribution of glutamate decarboxylase (GAD) and δ-amino butyric acid have been studied in the amygdaloid complex and in the stria terminalis system of the rat. The central and medial nuclei of the amygdala had significantly higher activities of GAD than the lateral olfactory tract nucleus, anterior amygdala, anterior lateral nucleus, posterior lateral nucleus, cortical nucleus, basomedial nucleus, basolateral nucleus, and pyriform cortex. The enzyme activity was about two and a half times higher in the central and medial nuclei than in the pyriform cortex. GABA was also significantly more concentratcd in these nuclei than in the pyriform cortex although this was not true for four other amino acids studied–glutamic acid, aspartic acid, taurine and glycine. GAD activity was also measured in the stria terminalis (the major afferent and efferent pathway of the amygdala) and in its bed nucleus. The enzyme activity was higher in the stria terminalis than in four other fibre tracts studied–the optic tract, anterior commissure, corpus callosum, and fimbria. GAD activity was exceptionally high in the bed nucleus of the stria terminalis particularly in its ventral part. The significance of the results are discussed in terms of what is known about the evolution and anatomy of the amygdala.     

Distribution of cholineacetyltransferase histochemistry in isthmus and medulla of Onchorynchus masu. Tract-tracing observation on the ascending meso-pontine cholinergic system

The distribution of cholinergic neurons was studies in the brain steam, medulla and rostral spinal cord of the salmon Onchorynchus masu using histochemical choline acetyltransferase (ChAT) detection. Cholinergic neurons were observed in the isthmus, cranial nerve motor nuclei and spinal cord. In order to characterize several cholinergic nuclei observed in the isthmus of O. masu, their projections were studied by application of 1,1'-dioctadecyl-3,3,3',3,'-tetramethylindocarbocyanine perchlorate (DiI) to selected structures of the brain. The secondary gustatory nucleus projected mainly to the lateral hypothalamic lobes, whereas the nucleus isthmi projected to the optic tectum and parvocellular superficial pretectal nucleus, as it was earlier described for the other teleost group. In addition, the other isthmic cholinergic nuclei in O. masu may be homologous to the meso-pontine system of mammals. We conclude that the cholinergic systems of teleosts show many primitive features that have been presented during evolution, together with exclusive to the group characteristics.     

Calcium-binding proteins and cytochrome oxidase activity in the turtle optic tectum with special reference to the tectofugal visual pathway

Using immunohistochemistry and a tracer technique we investigated the distribution in the optic tectum of turtles (Emys orbicularis and Testudo horsfieldi) of the calcium-binding proteins (CaBPr) parvalbumin (PV), calbindin (CB) and calretinin (CR) before and after labeling of the nucleus rotundus (Rot) with horseradish peroxidase. The optic tectum activity of the cytochrome oxidase (CO) was studied in parallel. In the principal link of the tectofugal visual pathway (central gray layer, SGC) in both chelonian species, the sparse PV-ir as well as CB- and CR-ir neurons were found significantly varying both in number and the intensity of immunoreactivity of their bodies and dendrites. In contrast, the superficial (SGFS) and deeper periventricular (SGP) tectal layers comprised numerous cells immunoreactive to all three CaBPr in different proportions. Only few retrogradely labeled tectorotundal SGC neurons expressed PV, CB or CR. The very large PV-ir neurons in SGC and SAC were not retrogradely labeled; morphologically they matched the efferent neurons with descending projections. SGC neurons of two chelonian species differed in the level of CO activity. Intense immunoreactivity to all three CaBPr and high CO activity were detected in both species in SGFS neuropil with some differences in sublaminar distribution patterns. The peculiarities of the CaBPr and CO activity distribution patterns in different segments of SGC neurons are discussed as related to the laminar organization of the turtle tectum and its retinal innervation. It is suggested that in the projection tectorotundal SGC neurons the CaBPr are concentrated mainly in their distal dendrites that contact retinal afferents in the superficial retinorecipient tectal layer.     

家鸽血管活性肠肽阳性神经元在顶盖和峡核中的分布及形态特征

用免疫细胞化学技术研究了血管活性肠肽(VIP)阳性神经元在家鸽(Columba livia)视顶盖和峡核中的分布,实验结果表明.在顶盖中VIP阳性神经元主要分布在Ⅱ_1层.并有少数VIP阳性神经元位于Ⅱ_1和Ⅲ层,在峡核大细胞部(Ime)和峡核小细胞部(Ipc),VIP阳性神经元基本呈均匀分布,在Imc和Ipc内分别占细胞总数的80%和50%.     

Glutamate Decarboxylase Distribution in Discrete Motor Nuclei in the Cat Brain

Abstract: The distribution of activity of glutamate decarboxylase (GAD), the enzyme synthesising γ-aminobutyric acid (GABA), was measured in the cat brain by means of microdissection of the structures from frozen slices and a radioisotopic assay for the enzyme. About 20 cerebral regions were chosen for study because of their role in sensorimotor integration. GAD presented an uneven distribution among these areas. Highest activities were found in the basal ganglia, particularly in the substantia nigra and in the globus pallidus, and to a lesser extent in the cerebellum. Relatively low levels of the enzyme were found in the thalamus and in the cerebral motor cortex. Special detailed studies were made in the caudate nucleus, the substantia nigra, and in the red nucleus for the purpose of defining the intranuclear distribution of their GABAergic innervation. There were only small differences in the rostro-caudal distribution of the enzyme in the head of the caudate nucleus but GAD activity was higher in the ventral than in the dorsal part of the structure. In the substantia nigra, GAD activity was high in both the medial and intermediate thirds of the structure. The GAD activity decreased from the caudal to the rostral part of the nucleus. GAD levels were lower in the caudal part of the red nucleus than in the rostral part. These results indicate that GABA would be present as a putative neurotransmitter in many motor nuclei of the cat brain. In view of the general inhibitory action of this amino acid, this could be related to the presence of inhibitory responses widely distributed in these nuclei as identified by mean of electrophysiological studies. The origin of these GABAergic innervations in many cases remains to be determined.     

Projections to the midbrain tectum in Salamandra salamandra L.

Following unilateral iontophoretic application of HRP into the optic tectum of Salamandra salamandra, retrogradely HRP-filled cells were found bilaterally in the pretectum, tegmentum isthmi, the reticular formation, pars medialis, and in the nucleus vestibularis magnocellularis. The area octavo-lateralis projects only to the caudal part of the tectum. Ipsilateral projections were noted from the dorsal gray columns of the cervical spinal cord, the dorsal tegmentum, the thalamus dorsalis pars medialis, thalamus dorsalis, pars anterior (to the rostral one-third of the tectum), the thalamus ventralis (in its entire rostro-caudal extent), and the preoptico-hypothalamic complex. Retrogradely filled cells were identified in deeper layers of the contralateral tectum. There are two telencephalic nuclei projecting ipsilaterally to the tectum via the lateral forebrain: the ventral part of the lateral pallium, and the posterior strioamygdalar complex.     

Primary retinal targets in the Atlantic loggerhead sea turtle,Caretta caretta

Summary Autoradiographic analysis distinguished twelve primary retinal targets in the diencephalon and the mesencephalon of the Atlantic loggerhead sea turtle, Caretta caretta. While the majority of fibers terminate contralaterally, sparse labelling is seen over ipsilateral thalamic nuclei. The dorsal optic nucleus is the most expansive retinal target in the dorsal thalamus. Four nuclei ventral and one dorsal, to the dorsal optic nucleus, receive retinal input. Before terminating in the optic tectum, labelled fibers pass through the pretectum terminating in four nuclei. Within the superficial zone of the optic tectum, three terminal zones are recognized. A distinct accessory tegmental tract separates from the main optic tract terminating in the basal optic nucleus.While such a multiplicity of retinal targets occurs among other reptiles, birds and mammals, it is presently impossible to accurately recognize visual homologies among amniotic vertebrates.     

The Topographical Distribution of Alanine, Aspartate, γ-Aminobutyric Acid, Glutamate, Glutamine, and Glycine in the Pigeon Optic Tectum and the Effect of Retinal Ablation

Abstract: The concentrations of alanine, aspartate, γ-aminobutyric acid, glutamine, glutamate, and glycine were measured in the pigeon optic nerve and in the individual tectal layers. Characteristic topographical distribution patterns were observed for the different amino acids. After unilateral retinal ablation, the concentration of aspartate and glutamate was decreased in the nerve and contralateral tectum. The reduction was restricted to the superficial part of the tectum, which receives a direct retinal input. The maximal loss was measured in the first two layers, where aspartate was reduced by 51% and glutamate by 75% in comparison with the ipsilateral side 4 weeks after ablation. The results favor a special role for aspartate and glutamate in pigeon retino-tectal afferents.     

Retinal projections in the catfish,Mystus vittatus (Bloch) as revealed by tracer studies with horseradish peroxidase

Summary The retinal efferents of the catfish, Mystus vittatus, were investigated with the use of the horseradish peroxidase (HRP) technique. Most retinal fibres extended contralateral to the eye that had received HRP label, while a few fascicles projected to the ipsilateral side without decussation in the optic chiasma. The contralateral fibres projected to the suprachiasmatic nucleus, the nucleus opticus dorsolateralis, the nucleus of the posterior commissure, the nucleus geniculatus lateralis, pretectal nuclear complex, and to two layers of the optic tectum, i.e., stratum fibrosum et griseum superficiale and stratum griseum centrale. The accessory optic tract arose from the inner area of the optic tract and extended ventromedially to the accessory optic nucleus. The ipsilateral fascicles projected to almost all the above mentioned nuclei, but these projections were comparatively sparse. The ipsilateral retinal projection was restricted to the rostral tectum.     

DISTRIBUTION OF CHOLINE ACETYLTRANSFERASE AND GLUTAMATE DECARBOXYLASE WITHIN THE SUBSTANTIA NIGRA AND IN OTHER BRAIN REGIONS FROM CONTROL AND PARKINSONIAN PATIENTS

—The distribution of choline acetyltransferase (ChAc, EC 2.3.1.6) and l -glutamate 1-carboxylyase (glutamate decarboxylase, GAD, EC 4.1.1.15) was studied in serial frontal slices of the substantia nigra (SN) (pars compacta, PC; pars reticulata, PR; an intermediate region, IR) as well as in other brain areas from post mortem tissue of control and Parkinsonian patients. Within the SN from control brain ChAc and GAD activities showed a distinctive distribution: ChAc activity in PC was higher than in PR and IR by 427% and 253% respectively and within PC the enzyme activity in the rostral part exceeded that in the control part by 353%. The GAD activity in PC was higher by 41% than that in PR and within PC seemed to be higher in the caudal than in the rostral part. For both enzyme activities there were no significant differences between PR and IR or within these regions. In Parkinsonian brain both ChAc and GAD activities were reduced to 15-25% of controls in all 3 regions of the SN. The distinctive distribution of ChAc and GAD activity found in the SN of control brain was abolished: no difference was observed between the 3 regions. However, within PC the ChAc activity was lower in the medial than in the rostral part. Since nigral ChAc is possibly located in interneurons, the decrease in enzyme activity may be connected with the cell loss observed in the SN of Parkinsonian brain. By contrast, nigral GAD is probably contained in terminals of strio-nigral neurons and the decrease in enzyme activity in Parkinson's disease in the absence of striatal cell loss, may reflect a change in the functional state of these GABA neurons. Among various areas of control brains ChAc activity was highest in caudate nucleus and putamen while GAD was highest in SN. caudate nucleus, putamen and cerebral cortex. In Parkinsonian brain the most severe reduction in ChAc and GAD activities was found in the SN.     

The organization of cholinergic neurons in the mesencephalon of the eel,Anguilla anguilla,as determined by choline acetyltransferase immunohistochemistry and acetylcholinesterase enzyme histochemistry

Cholinergic systems in the midbrain of the eel were identified by using histochemical procedures for the demonstration of the enzymes choline acetyltransferase (ChAT) and acetylcholinesterase. Neurons detected by both methods are located in the stratum periventriculare of the tectum, cranial motor nuclei III and IV, nucleus isthmi, nucleus gustatorius secundarius, nucleus reticularis superior, and nucleus lateralis valvulae. Some projections of these cell groups were studied by injecting horseradish peroxidase into selected brain regions. Cholinergic neurons make up about 10% of the neurons in the stratum periventriculare of the tectum and are a subset of the type-XIV neurons. Neurons in n. isthmi project primarily to the ipsilateral tectum; some cholinergic isthmal neurons project to n. pretectalis superficialis. A few ChAT-positive axons, perhaps belonging to the tectopetal system, were observed in the optic nerve. The cholinergic neurons of n. gustatorius secundarious project to the inferior lobes of the hypothalamus. The neurons of the superior reticular nucleus are a cholinergic subset of the superior reticular formation. Their axons project rostrally, probably to the thalamus and pretectum. The findings are discussed in relation to functional features of the mesencephalon, particularly in relation to locomotory control.     

GABA IN THE OLFACTORY BULB AND OLFACTORY NUCLEUS OF THE RAT: THE DISTRIBUTION OF GAMMA-AMINOBUTYRIC ACID, GLUTAMIC ACID DECARBOXYLASE, GABA TRANSAMINASE AND SUCCINATE SEMIALDEHYDE DEHYDROGENASE

Abstract— The distribution of GABA and enzymes involved in its metabolism was investigated in the different regions of the olfactory bulb and olfactory nucleus. The highest levels of GABA in the olfactory bulb were found in the layers rich in nerve terminals (31 μmol/g dry wt.). A similar distribution was found in the olfactory nucleus although the overall level of GABA was only a quarter of that measured in the bulb. Glutamic acid decarboxylase (GAD) levels in the various layers of the olfactory nucleus were similar in distribution to those of GABA. However, the correlation between GAD and GABA did not hold for the olfactory bulb, particularly in the granule cell layer and the medulla. The activities of GAD and the levels of GABA are significantly higher in the bulb than in the nucleus but succinic acid scmialdehyde dehydrogenase and GABA aminotransaminase activities are almost identical in both regions.     

Neurochemical characteristics of the turtle optic tectum: comparison with other reptilian species and birds

Data on distribution of biologically active substances in the turtle optic tectum are compared with results of similar experiments on other reptilian as well as on avian species. In two turtle species (Testudo horsfield and Emys orbicularis), immunoreactivity to monoamines (5-HT and TH), NPY, as well as NADPH-d activity were similarly distributed in neuropil of the SGFS retinorecipient part and in that of the SGP/SAP periventricular layers. Immunoreactivity to neuropeptides SP and m-Enk was maximal in neuropil of the SGFS non-retinorecipient part. The periventricular layers were characterized by the abundant radial SP- and mENK-ir as well as the NADPH-d-positive neurons. Diffusely dispersed ChAT-ir elements and many ir fibers perpenducilar to the tectal surface were observed in the SGFS retinorecipient part; the SGFS non-retinorecipient part contained a dense plexus of thick ir fibers and diffusely distributed ir terminals. The GABA ir cells were the most numerous in the tectum; they were spread in all tectal layers. Thus, various biologically active substance located in superficial retinorecipient tectal sublayers could affect processing and transmission of information via ascending dendrites of neurons in deeper layers. The cells containing SP, m-Enk, and NADPH-d had laminar organization in SGP; via the system of ascending and descending axons, they are able to affect other structures within and outside of the optic tectum. Putative sources of tectal modulatory innervation are discussed. In all studied reptilian and avian species, the principal similarity is revealed in the neurochemical organization. Some differences might be explained by the level of tectal differentiation due to factors of phylogenetic evolution and/or adaptive specialization.     

Brain-derived neurotrophic factor (BDNF)-like immunoreactivity localization in the retina and brain of Cichlasoma dimerus (Teleostei, Perciformes)

Brain-derived neurotrophic factor (BDNF) is a neurotrophin involved in the development and maintenance of vertebrate nervous systems. Although there were several studies in classical animal models, scarce information for fish was available. The main purpose of this study was to analyze the distribution of BDNF in the brain and retina of the cichlid fish Cichlasoma dimerus. By immunohistochemistry we detected BDNF-like immunoreactive cells in the cytoplasm and the nuclei of the ganglion cell layer and the inner nuclear layer of the retina. In the optic tectum, BDNF-like immunoreactivity was detected in the nucleus of neurons of the stratum periventriculare and the stratum marginale and in neurons of the intermediate layers. In the hypothalamus we found BDNF-like immunoreactivity mainly in the cytoplasm of the nucleus lateralis tuberis and the nucleus of the lateral recess. To confirm the nuclear and cytoplasm localization of BDNF we performed subcellular fractionation, followed by Western blot, detecting a 39 kDa immunoreactive-band corresponding to a possible precursor form of BDNF in both fractions. BDNF-like immunoreactivity was distributed in areas related with photoreception (retina), the integration center of retinal projections (optic tectum) and the control center of background and stress adaptation (hypothalamus). These results provide baseline anatomical information for future research about the role of neurotrophins in the adult fish central nervous system.     

Neurochemical characteristics of the turtle optic tectum: Comparison with other reptilian species and birds

Data on distribution of biologically active substances in the turtle optic tectum are compared with results of similar experiments on other reptilian as well as on avian species. In two turtle species (Testudo horsfield and Emys orbicularis), immunoreactivity to monoamines (5-HT and TH), NPY, as well as NADPH-d activity were similarly distributed in neuropil of the SGFS retinorecipient part and in that of the SGP/SAP periventricular layers. Immunoreactivity to neuropeptides SP and m-Enk was maximal in neuropil of the SGFS non-retinorecipient part. The periventricular layers were characterized by the abundant radial SP- and mENK-ir as well as the NADPH-d-positive neurons. Diffusely dispersed ChAT-ir elements and many ir fibers perpenducilar to the tectal surface were observed in the SGFS retinorecipient part; the SGFS non-retinorecipient part contained a dense plexus of thick ir fibers and diffusely distributed ir terminals. The GABA ir cells were the most numerous in the tectum; they were spread in all tectal layers. Thus, various biologically active substances located in superficial retinorecipient tectal sublayers could affect processing and transmission of information via ascending dendrites of neurons in deeper layers. The cells containing SP, m-Enk, and NADPH-d had laminar organization in SGP; via the system of ascending and descending axons, they are able to affect other structures within and outside of the optic tectum. Putative sources of tectal modulatory innervation are discussed. In all studied reptilian and avian species, the principal similarity is revealed in the neurochemical organization. Some differences might be explained by the level of tectal differentiation due to factors of phylogenetic evolution and/or adaptive specialization.     

Choline acetyltransferase, glutamic acid decarboxylase and somatostatin in the kainic acid model for chronic temporal lobe epilepsy

The aim of the study was to investigate neurochemical changes in a kainic acid (KA; 10 mg/kg, s.c.)-induced spontaneous recurrent seizure model of epilepsy, 6 months after the initial KA-induced seizures. The neuronal markers of cholinergic and gamma-aminobutyric acid (GABA)ergic systems, i.e. choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) activities, and a marker for neuropeptide, i.e. level of somatostatin, have been investigated. The brain regions investigated were the hippocampus, amygdala/piriform cortex, caudate nucleus, substantia nigra and the frontal, parietal, temporal and occipital cortices. Six months after KA injection, reduced ChAT activity was observed in the amygdala/piriform cortex (47% of control; p<0.001), increased ChAT activity in the hippocampus (119% of control; p<0.01) and normal ChAT activity in the other brain regions. The activity of GAD was significantly increased in all analysed cortical regions (between 146 and 171% of control), in the caudate nucleus (144% of control; p<0.01) and in the substantia nigra (126% of control; p<0.01), whereas in the amygdala/piriform cortex, the GAD activity was moderately lowered. The somatostatin level was significantly increased in all cortical regions (between 162 and 221% of control) as well as in the hippocampus (119% of control), but reduced in the amygdala/piriform cortex (45% of control; p<0.01). Six months after KA injection, the somatostatin:GAD ratio was lowered in the amygdala/piriform cortex (49% of control) and in the caudate nucleus (41% of control), whereas it was normal in the hippocampus and moderately increased in the cortical brain regions. A positive correlation was found between seizure severity and the reduction of both ChAT activities and somatostatin levels in the amygdala/piriform cortex. The results show a specific pattern of changes for cholinergic, GABAergic and somatostatinergic activities in the chronic KA model for epilepsy. The revealed data suggest a functional role for them in the new network that follows spontaneous repetitive seizures.     

A crossed projection from the optic tectum to craniocervical premotor areas in the brainstem reticular formation. An anterograde and retrograde tracing study in the mallard (Anas platyrhynchos L.)

The optic tectum in birds receives visual information from the contralateral retina. This information is passed through to other brain areas via the deep layers of the optic tectum. In the present study the crossed tectobulbar pathway is described in detail. This pathway forms the connection between the optic tectum and the premotor area of craniocervical muscles in the contralateral paramedian reticular formation. It originates predominantly from neurons in the ventromedial part of stratum griseum centrale and to a lesser extent from stratum album centrale. The fibers leave the tectum as a horizontal fiber bundle, and cross the midline through the caudal radix oculomotorius and rostral nucleus oculomotorius. On the contralateral side fibers turn to ventral and descend caudally in the contralateral paramedian reticular formation to the level of the obex. Labeled terminals are found in the ipsilateral medial mesencephalic reticular formation lateral to the radix and motor nucleus of the oculomotor nerve, and in the contralateral paramedian reticular formation, along the descending tract. Neurons in the medial mesencephalic reticular formation in turn project to the paramedian reticular formation. Through the crossed tectobulbar pathway visual information can influence the activity of craniocervical muscles via reticular premotor neurons.     

北草蜥中脑视叶组织学结构观察

用光学显微镜对北草蜥(Takydromus septentrionalis)中脑视叶组织学结构进行了观察。视叶分为背侧的顶盖和腹侧的被盖,两者无明显界限。顶盖处灰质和白质交替排列。由表及里,可分为分子层、外灰质层、浅白质层、中灰质层、中白质层、深灰质层、深白质层和室管膜层。被盖处细胞层次不明显。在视叶的前部有横行的纤维将左右视叶联系起来。左右视叶室与中脑水管以及两视叶间的纵沟在视叶的中、后部相通。同时将北草蜥的中脑与其它低等脊椎动物进行了比较,推测北草蜥在进化上处于较低等地位。     

Differential sensitivity of cholinergic and GABAergic neurons in chick embryos treated intracerebrally with ethanol at 8 days of embryonic age

We have shown that in embryos treated with ethanol in ovo during days 1–3, a critical period of neuroembryogenesis, cholinergic neuronal phenotypic expression is decreased whereas GABAergic and catecholaminergic neuronal populations are increased as assessed by neuronal markers choline acetyltransferse (ChAT), glutamic acid decarboxylase (GAD) and tyrosine hydroxylase (TH) respectively. In this study, ethanol was administered intracerebrally to embryos at embryonic day 8, embryos were sacrificed at day 9 and ChAT and GAD activities assayed separately in cerebral hemispheres and remaining brain (diencephalon-midbrain and optic lobes). We found that ChAT activity was enhanced in the cerebral hemispheres only, whereas GAD activity was decreased in both cerebral hemispheres and remaining brain. We have concluded that the differential responses of neuronal phenotypes to ethanol may reflect compensatory mechanisms to ethanol insult. Moreover, these findings emphasize the vulnerability of the GABAergic neuronal phenotypes to ethanol neurotoxicity during early brain development in the chick.