Prevalence and risk factors for colonization of Clostridium difficile among adults living near livestock farms in the Netherlands.

A cross-sectional study was performed among 2494 adults not living or working on a farm to assess prevalence of Clostridium difficile (CD) colonization and risk factors in a livestock dense area. CD prevalence was 1·2%. Twenty-one persons were colonized with a toxigenic strain and nine with a non-toxigenic strain. CD-positive persons did not live closer to livestock farms than individuals negative for CD. Antibiotic exposure in the preceding 3 months was a risk factor for CD colonization (odds ratio 3·70; 95% confidence interval 1·25-10·95).

Extended-spectrum ß-lactamase- and pAmpC-producing Enterobacteriaceae among the general population in a livestock-dense area.

OBJECTIVES: In the Netherlands there is an ongoing debate regarding environmental health risks of livestock farming for neighbouring residents. This explorative study aims to determine the prevalence of carriage of extended-spectrum ß-lactamase and/or plasmid-mediated AmpC-producing Enterobacteriaceae (ESBL/pAmpC-E) in the general population living in a livestock-dense area, and to study associations between determinants, including exposure through contact with animals and the environment, and human carriage of ESBL/pAmpC-E. METHODS: A cross-sectional study was performed among 2432 adults (aged 20-72 years) in 12 temporary research centres in the south of the Netherlands, consisting of a questionnaire and analysis of a faecal sample to assess carriage of ESBL/pAmpC-E. Risk factors were analysed using logistic regression. RESULTS: The prevalence for carriage of ESBL/pAmpC-E was 4.5% (109/2432; 95% CI 3.7-5.4) ranging from 1.4% to 10.9% among the research centres. ESBL/pAmpC resistance genes were detected in Escherichia coli and Klebsiella pneumoniae isolates obtained from these 109 persons and the most common ESBL-resistance genes were blaCTX-M-15, blaCTX-M-14/17 and blaCTX-M-1, originating from 76 participants. Travel in the previous 12 months to Africa, Asia or Latin America (OR 2.82; 95% CI 1.71-4.63), having kept cows for a hobby in the previous 5 years (OR 3.77; 95% CI 1.22-11.64), usage of proton-pump inhibitors (OR 1.84; 95% CI 1.05-3.23), and living within 1000 m of a mink farm (OR 2.26; 95% CI 1.28-3.98) were identified as risk factors. Exposure to poultry was not identified as a risk factor. CONCLUSIONS: Overall, living in close proximity to livestock animals and farms does not seem to be a risk factor for carriage of ESBL/pAmpC-E.

The distribution of GABA-containing perikarya, fibers, and terminals in the forebrain and midbrain of pigeons, with particular reference to the basal ganglia and its projection targets.

Immunohistochemical techniques were used to study the distributions of glutamic acid decarboxylase (GAD) and gamma-aminobutyric acid (GABA) in pigeon forebrain and midbrain to determine the organization of GABAergic systems in these brain areas in birds. In the basal ganglia, numerous medium-sized neurons throughout the striatum were labeled for GABA, while pallidal neurons, as well as a small population of large, aspiny striatal neurons, labeled for GAD and GABA. GAD+ and GABA+ fibers and terminals were abundant throughout the basal ganglia, and GABAergic fibers were found in all extratelencephalic targets of the basal ganglia. Most of these targets also contained numerous GABAergic neurons. In pallial regions, approximately 10-12% of the neurons were GABAergic. The outer rind of the pallium was more intensely labeled for GABAergic fibers than the core. The olfactory tubercle region, the ventral pallidum, and the hypothalamus were extremely densely labeled for GABAergic fibers, while GABAergic neurons were unevenly distributed in the hypothalamus. GABAergic neurons and fibers were abundant in the dorsalmost part of thalamus and the dorsal geniculate region, while GABAergic neurons and fibers were sparse (or lightly labeled) in the thalamic nuclei rotundus, triangularis, and ovoidalis. Further, GABAergic neurons were abundant in the superficial tectal layers, the magnocellular isthmic nucleus, the inferior colliculus, the intercollicular region, the central gray, and the reticular formation. GABAergic fibers were particularly abundant in the superficial tectal layers, the parvocellular isthmic nucleus, the inferior colliculus, the intercollicular region, the central gray, and the interpeduncular nucleus. These results suggest that GABA plays a role as a neurotransmitter in nearly all fore- and midbrain regions of birds, and in many instances the observed distributions of GABAergic neurons and fibers closely resemble the patterns seen in mammals, as well as in other vertebrates.

Organization of the avian "corticostriatal" projection system: a retrograde and anterograde pathway tracing study in pigeons.

Birds have well-developed basal ganglia within the telencephalon, including a striatum consisting of the medially located lobus parolfactorius (LPO) and the laterally located paleostriatum augmentatum (PA). Relatively little is known, however, about the extent and organization of the telencephalic "cortical" input to the avian basal ganglia (i.e., the avian "corticostriatal" projection system). Using retrograde and anterograde neuroanatomical pathway tracers to address this issue, we found that a large continuous expanse of the outer pallium projects to the striatum of the basal ganglia in pigeons. This expanse includes the Wulst and archistriatum as well as the entire outer rind of the pallium intervening between Wulst and archistriatum, termed by us the pallium externum (PE). In addition, the caudolateral neostriatum (NCL), pyriform cortex, and hippocampal complex also give rise to striatal projections in pigeon. A restricted number of these pallial regions (such as the "limbic" NCL, pyriform cortex, and ventral/caudal parts of the archistriatum) project to such ventral striatal structures as the olfactory tubercle (TO), nucleus accumbens (Ac), and bed nucleus of the stria terminalis (BNST). Such "limbic" pallial areas also project to medialmost LPO and lateralmost PA, while the hyperstriatum accessorium portion of the Wulst, the PE, and the dorsal parts of the archistriatum were found to project primarily to the remainder of LPO (the lateral two-thirds) and PA (the medial four-fifths). The available evidence indicates that the diverse pallial regions projecting to the striatum in birds, as in mammals, are parts of higher order sensory or motor systems. The extensive corticostriatal system in both birds and mammals appears to include two types of pallial neurons: 1) those that project to both striatum and brainstem (i.e., those in the Wulst and the archistriatum) and 2) those that project to striatum but not to brainstem (i.e., those in the PE). The lack of extensive corticostriatal projections from either type of neuron in anamniotes suggests that the anamniote-amniote evolutionary transition was marked by the emergence of the corticostriatal projection system as a prominent source of sensory and motor information for the striatum, possibly facilitating the role of the basal ganglia in movement control.

Evidence for a possible avian dorsal thalamic region comparable to the mammalian ventral anterior, ventral lateral, and oral ventroposterolateral nuclei.

In the present study, we investigated whether a dorsal thalamic region comparable to the motor part of the mammalian ventral tier (the ventral anterior nucleus, the ventral lateral nucleus, and the oral ventroposterolateral nucleus) exists in pigeon. With this aim, we reinvestigated the projections of the pigeon dorsal pallidum to the dorsal thalamus by using 1) injections of the anterogradely transported form of biotinylated dextran amine (BDA; 10,000 molecular weight) in the pigeon dorsal pallidum (paleostriatum primitivum) and 2) injections of the retrogradely transported form of BDA (3,000 molecular weight) in the pigeon dorsal thalamus. Our results indicate that the dorsal pallidum in pigeons projects to three areas of the dorsal thalamus: the dorsointermediate posterior nucleus, the ventrointermediate area, and the nucleus subrotundus. Only the projection to the dorsointermediate posterior nucleus was described previously (Karten and Dubbeldam [1973] J. Comp. Neurol. 148:61-90; Kitt and Brauth [1982] Neuroscience 6:1551-1566). To investigate whether any of the dorsal thalamic nuclei receiving pallidal input project to a motor cortical field, injections of the retrograde tracer Fluoro-Gold were placed into the rostral Wulst. This is an avian cortical field that appears to contain a region comparable to mammalian primary somatomotor cortex (Karten [1971] Anat. Rec. 169:353; Wild [1992] J. Comp. Neurol. 287:1-18). Our results indicate that neurons in the rostral ventrointermediate area, but not in the nucleus subrotundus, the dorsointermediate posterior nucleus, or the intermediate or caudal parts of the ventrointermediate area, project to the rostral Wulst. In addition to the input from the dorsal pallidum, the avian ventrointermediate area also receives input from the lateral substantia nigra and the lateral and internal cerebellar nuclei (present results). Our results suggest the existence in birds of a pallidothalamocortical loop similar to the pallidoventral tier-motor cortex loop of mammals and suggest that the avian ventrointermediate area is comparable to the motor part of the mammalian ventral tier in both location and connections. If this is confirmed by physiological experiments, then it would indicate that basal ganglia control of movement mediated by a pallidothalamocortical loop may have evolved with the stem reptiles.

Distributions of GABAA, GABAB, and benzodiazepine receptors in the forebrain and midbrain of pigeons.

Autoradiographic and immunohistochemical methods were used to study the distributions of GABAA, GABAB and benzodiazepine (BDZ) receptors in the pigeon fore- and midbrain. GABAA, GABAB and BDZ binding sites were found to be abundant although heterogeneously distributed in the telencephalon. The primary sensory areas of the pallium of the avian telencephalon (Wulst and dorsal ventricular ridge) tended to be low in all three binding sites, while the surrounding second order belt regions of the pallium were typically high in all three. Finally, the outermost rind of the pallium (termed the pallium externum by us), which surrounds the belt regions and projects to the striatum of the basal ganglia, was intermediate in all three GABAergic receptors types. Although both GABAA and benzodiazepine receptors were abundant within the basal ganglia, GABAA binding sites were densest in the striatum and BDZ binding sites were densest in the pallidum. Among the brainstem regions receiving GABAergic basal ganglia input, the anterior and posterior nuclei of the ansa lenticularis showed very low levels of all three receptors, while the lateral spiriform nucleus and the ventral tegmental area/substantia nigra complex contained moderate abundance of the three binding sites. The dorsalmost part of the dorsal thalamus (containing nonspecific nuclei) was rich in all three binding sites, while the more ventral part of the dorsal thalamus (containing specific sensory nuclei), the ventral thalamus and the hypothalamus were poor in all three binding sites. The pretectum was also generally poor in all three, although some nuclei displayed higher levels of one or more binding sites. The optic tectum, inferior colliculus, and central gray were rich in all three sites, while among the isthmic nuclei, the parvicellular isthmic nucleus was conspicuously rich in BDZ sites. The results show a strong correlation of the regional abundance of GABA binding sites with previously described distributions of GABAergic fibers and terminals in the avian forebrain and midbrain. The regional distribution of these binding sites is also remarkably similar to that in mammals, indicating a conservative evolution of forebrain and midbrain GABA systems among amniotes.

Cellular expression of ionotropic glutamate receptor subunits on specific striatal neuron types and its implication for striatal vulnerability in glutamate receptor-mediated excitotoxicity.

Glutamate receptors are composed of subtype-specific subunits. Variation in the precise subunit composition of a receptor may result in significant functional differences. Thus, a precise knowledge of subunit composition on striatal neurons is a prerequisite for understanding the selective vulnerability of striatal neurons to excitatory amino acids. In the present study, we used an immunohistochemical double-labelling approach to localize ionotropic glutamate receptor subunits (NMDAR1, GluR1, GluR2/3, GluR4 and GluR5/6/7) on specific striatal neuron populations. Our results showed that striatal cholinergic and somatostatin interneurons were not labelled for the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate, receptor subunits GluR1, GluR2/3 and GluR4. Most cholinergic and somatostatin interneurons (83.3% to 100%), however, were double-labelled for the N-methyl-D-aspartate receptor subunit NR1 and kainic acid receptor subunits GluR5/6/7. All parvalbumin interneurons were labelled for GluR1 and GluR4, and 96% GluR1 positive and 95% GluR4 positive neurons were also double-labelled as parvalbumin interneurons. About half of all parvalbumin interneurons co-localized with GluR2/3, and over 97% were labelled for NR1 and GluR5/6/7. Among striatal projection neurons, enkephalin-positive (mainly striatopallidal) neurons, striatonigral neurons (mainly substance P-positive) and calbindin-positive matrix neurons were not immunostained for GluR1 or GluR4. In contrast, 95% to 100% of each of these types of projection neurons were double-labelled for NR1, GluR2/3 and GluR5/6/7. Our results demonstrate that striatal neuron types differ in their expression of ionotropic glutamate receptor subunits and subtypes. The clear difference between striatal interneurons and projection neurons in ionotropic glutamate receptor subtypes/subunits supports the idea that differential glutamate receptor expression mechanism may account for the selective vulnerability of striatal projection neurons to excitotoxicity, and that glutamate receptor-mediated excitotoxicity may be involved in the striatal neurodegenerative diseases.

The effects of radiofrequency lesion or transection of the fimbria-fornix on latent inhibition in the rat.

Latent inhibition consists of a decrement in conditioning to a stimulus as a result of its prior non-reinforced pre-exposure. Based on evidence pointing to the involvement of the hippocampus and the nucleus accumbens in latent inhibition disruption, it has been proposed that latent inhibition depends on the integrity of the subicular input to the nucleus accumbens. Since fibers originating in the subiculum and destined for the nucleus accumbens run through the fimbria-fornix, we assessed the effects of radiofrequency lesion or transection of the fimbria-fornix, on latent inhibition. The effectiveness of both lesions was demonstrated by the total disappearance of acetylcholinesterase staining in the hippocampus and of retrogradely labeled cells in the hippocampus/subiculum following the injection of the retrograde tracer biotin-dextran amine into the shell subregion of the nucleus accumbens. Likewise, in accord with previously documented behavioral effects of lesions to the hippocampus and related structures, both lesions increased spontaneous activity and disrupted performance in Morris water maze, and the radiofrequency lesion facilitated the acquisition of two-way active avoidance. In spite of the above, latent inhibition remained unaffected by both fimbria-fornix lesions, indicating that the critical projections subserving latent inhibition are not those traversing the fimbria-fornix from the hippocampus/subiculum to the nucleus accumbens. The implications of these results for the neural circuitry of latent inhibition and the latent inhibition model of schizophrenia are discussed.

The anatomical substrate for telencephalic function.

The basic thesis for this study was that the telencephalon is needed to make decisions in new situations. Subsidiary hypotheses were that the telencephalon consists of: (a) a sensorimotor system which generates motor activity from sensory input and (b) a selection system which makes choices from possible motor programs. It was postulated that the selection system should fulfil the following requirements: be accessible for past and present events, have the capacity to process this information in a nondetermined way with a possibility for ordering, and have access to motor-affecting systems (the sensorimotor system). The ability of the selection system to correlate information in a nonpredetermined way was considered most important. In short: The selection system should be able to associate any information in any combination, and have the capability for internal control of neuronal activity and external selection of motor programs (see Fig. 1A.) Xenopus laevis was chosen as a subject, since it has a relatively simple telencephalon, with characteristics that it shares with "primitive" species of different vertebrate classes, and because it is easy to maintain as a laboratory animal. The main method used was the determination of connections with HRP. The pallium was in the focus of attention, since it was considered to be the core of the selection system. Immunohistochemistry was used as an additional parameter to compare Xenopus laevis forebrain with those of other vertebrates. The results showed that the pallium can be subdivided into a rostral (third) and a caudal (two-thirds) entity. The rostral third is the main recipient for thalamic and olfactory input. The caudal two-thirds are linked up to the rostral third and have a refined microcircuitry. Efferents from the pallium remain restricted to the forebrain. The entire pallium consists of a network of intrinsic reciprocal connections and can be considered to be positioned between the medial pallium (hippocampus), septum, and amygdaloid complex (amygdala). As a whole this system targets the hypothalamus. The hypothalamus in turn projects into the striatum complex (striatum with anterior entopeduncular nucleus). The rostral dorsal pallium and the amygdaloid complex also project into the striatum complex. The striatum is positioned between the sensory input from the thalamus and olfactory bulbs, and the motor output to the medulla. It is concluded, on the basis of its straightforward input-output relations and uniform appearance, that the striatum complex fulfils the requirements for a sensorimotor system. The pallium together with the septum, amygdaloid complex, and hypothalamus fulfils the requirements for a selection system.(ABSTRACT TRUNCATED AT 400 WORDS)

Thalamostriatal projection neurons in birds utilize LANT6 and neurotensin: a light and electron microscopic double-labeling study.

Based on its location, connectivity and neurotransmitter content, the dorsal thalamic zone in birds appears to be homologous to the intralaminar, midline, and mediodorsal nuclear complex in the thalamus of mammals. We investigated the neuroactive substances used by thalamostriatal projection neurons of the dorsal thalamic zone in the pigeon. Single-labeling experiments showed that many neurons in the dorsal thalamic zone are immunoreactive for neurotensin and the neurotensin-related hexapeptide, (Lys8,Asn9)NT(8-13) (LANT6). Double-labeling experiments, using the retrograde fluorescent tracer, FluoroGold, combined with fluorescence immunocytochemistry for either LANT6 or neurotensin, showed that neurotensin- and LANT6-containing neurons in the dorsal thalamic zone project to the striatum of the basal ganglia. Immunofluorescence double-labeling experiments showed that neurotensin and LANT6 are often (possibly always) co-expressed in neurons in the dorsal thalamic zone. Electron microscopic immunohistochemical double-labeling showed that LANT6 terminals in the striatum make asymmetric contacts with heads of spines labeled for substance P and heads of spines not labeled for substance P, suggesting that these terminals synapse with both substance P-containing and non-substance P-containing medium spiny striatal projection neurons. These findings indicate that LANT6 and neurotensin may be utilized as neurotransmitters in thalamostriatal projections in birds and raise the possibility that this may also be the case in other amniotes.

The effects of hippocampal and fimbria-fornix lesions on prepulse inhibition.

The present experiments tested the effects of conventional (dorsal aspiration and electrolytic) and excitotoxic (N-methyl-D-aspartate [NMDA]) hippocampal lesions and fimbria-fornix (FF) transection on prepulse inhibition (PPI) of startle response and on open-field activity. Activity was increased by FF transection and by conventional but not excitotoxic hippocampal lesions; complete NMDA lesion increased amphetamine-induced activity. Whereas dorsal hippocampal aspiration lesion disrupted PPI, the phenomenon was not affected by dorsal hippocampal electrolytic lesion, partial or complete excitotoxic (NMDA) hippocampal lesions, or complete FF transection, which interrupted the cholinergic input to the hippocampus as well as the hippocampal-subicular input to the nucleus accumbens. Systemic apomorphine disrupted PPI in both FF-transected rats and their controls. It is suggested that the hippocampus is essential for PPI disruption rather than for PPI expression.

Biotinylated dextran amine as an anterograde tracer for single- and double-labeling studies.

Fluorescent dextran amines have recently been reported to be useful for anterograde pathway tracing. However, fluorescent markers are not always ideal for detailed mapping studies. We therefore evaluated the efficacy of a biotinylated dextran amine (BDA) for anterograde labeling in several different preparations. BDA was visualized with an avidin-biotinylated HRP (ABC) procedure followed by a standard or metal-enhanced diaminobenzidine (DAB) reaction. After iontophoretic injections of BDA into neocortex-like telencephalic regions in pigeons or into visual or somatosensory cortex in rats, there was excellent and abundant labeling of axons and terminals in forebrain, midbrain and hindbrain target areas with 1-week survival times. Large pressure injections of BDA into the avian telencephalon were also found to result in extensive anterograde labeling. We then carried out a series of studies using 2-color DAB double-labeling to determine effective approaches for combining BDA labeling with other labeling methods. Using an isolated embryonic chick spinal cord-hindlimb preparation, we combined BDA labeling with another anterograde labeling method to differentially label two sets of projections. In these studies, sensory neuron and motoneuron projections into the limb from the same segmental level, or motoneuron projections into the limb from two separate segments were differentially labeled by using HRP (visualized first with a blue/black metal-DAB reaction) and BDA (visualized second with a brown DAB reaction). In other double-labeling studies, we combined BDA labeling of axons and terminals with immunohistochemical labeling of neurons. In these experiments, telencephalic neurons in pigeons or rats were labeled immunohistochemically for parvalbumin or substance P (using a brown DAB reaction) and BDA-labeled axons were labeled blue/black (using a metal-intensified DAB reaction). Double-labeling was successful regardless of whether the entire immunohistochemical labeling procedure preceded or followed the BDA labeling procedure. Together, these studies show that BDA is effective for anterograde pathway tracing and can be used in double-label studies with other labeling methods.

Pathway tracing using biotinylated dextran amines.

Biotinylated dextran amines (BDA) are highly sensitive tools for anterograde and retrograde pathway tracing studies of the nervous system. BDA can be reliably delivered into the nervous system by iontophoretic or pressure injection and visualized with an avidin-biotinylated HRP (ABC) procedure, followed by a standard or metal-enhanced diaminobenzidine (DAB) reaction. High molecular weight BDA (10 k) yields sensitive and exquisitely detailed labeling of axons and terminals, while low molecular weight BDA (3 k) yields sensitive and detailed retrograde labeling of neuronal cell bodies. The detail of neuronal cell body labeling can be Golgi-like. BDA tolerates EM fixation and processing well and can, therefore, be readily used in ultrastructural studies. Additionally, BDA can be combined with other anterograde or retrograde tracers (e.g. PHA-L or cholera toxin B fragment) and visualized either by multi-color DAB multiple-labeling - if permanent labels are desired, or by using multiple simultaneous immunofluorescence - if fluorescence viewing is desired. In the same manner, BDA pathway tracing and neurotransmitter immunolabeling can be combined. Note that BDA pathway tracing can also be combined with anterograde or retrograde labeling with fluorescent dextran amines, if one wishes to exclusively use tracers with the favorable transport properties and sensitivities of dextran amines. In this case, the BDA can be visualized together with the fluorescent dextran amines using fluorescence labeling for the BDA, or the fluorescent dextran amines can be visualized together with the BDA by multicolor DAB labeling via immunolabeling of the fluorescent dextran amines using anti-fluorophore antisera. BDA is, thus, a flexible and valuable pathway tracing tool that has gained widespread popularity in recent years.

Ultrastructural morphology of synapses formed by corticostriatal terminals in the avian striatum.

We studied the ultrastructural morphology of corticostriatal projections from two different avian 'neocortical' regions, namely, the hyperstriatum accessorium (HA) and the pallium externum (PE). Biotinylated dextran amine (BDA) was used to label the corticostriatal projection from either HA or PE to the striatum. The corticostriatal axons from both the PE and HA possessed numerous beaded varicosities with the striatum. These varicosities were filled with numerous round vesicles characterizing them as terminals. These terminals formed asymmetric synapses with spine heads and with dendrites of striatal neurons. The axospinous synapses outnumbered the axodendritic synapses by more than two to one. The diameters of labeled axons were typically 250-500 nm. The labeled terminals were typically 400-750 nm in diameter. No obvious differences between the ultrastructural morphology of the HA and the PE corticostriatal projections were observed. These data show that corticostriatal terminals and their synaptic contacts in birds are similar to those described in mammals.

Interspecific differences in the expression of the AMPA-type glutamate receptors and parvalbumin in the nucleus of Edinger-Westphal of chicks and pigeons.

The distribution of AMPA-type glutamate receptor (GluR) subunits was studied in the Edinger-Westphal nucleus (EW) of chicks and pigeons. GluR1, GluR2, GluR3 and GluR4 subunits appeared to be present in EW neurons of both species, but interspecific differences were observed in the abundance of the different types of subunits found in EW neurons. Of particular note, GluR2 immunoreactivity was present in the vast majority (ca. 80%) of neurons of pigeon EW but was found in only a small fraction (ca. 15%) of chick EW neurons. Scarcity of the GluR2 subunit in chick EW was confirmed by in situ hybridization. Because of the tendency for parvalbumin to be localized to neurons that are selectively deficient in GluR2, we also studied the localization of parvalbumin, as well as other calcium-binding proteins, in EW of chick and pigeon. Parvalbumin was found in more than 50% of chick EW neurons but was not detected in pigeon EW neurons. Our results suggest that there are major glutamatergic inputs to EW neurons in both pigeons and chicks. Furthermore, there are likely to be more AMPA-type calcium-permeable glutamate receptors in EW neurons of chick than in pigeon, since it is known that the subtype containing the edited GluR2 subunit is not calcium permeable.

Comparisons of the densities of NADPHd reactive and nNOS immunopositive neurons in the hippocampus of three age groups of young nonhandled and handled rats.

The complete absence of handling of male rats during neonatal development (from birth to postnatal day 21) correlates with an impairment of latent inhibition [J. Feldon, I. Weiner, From an animal model of an attentional deficit towards new insights into the pathophysiology of schizophrenia, J. Psychiatr. Res. 26 (1992) 345-366.]. Such nonhandling of rats reportedly also correlates with a decreased expression of reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd) reactivity in the hippocampus in adult rats (6 months of age) when compared with rats of the same age that were handled during the same neonatal period [R.R. Vaid, B.K. Yee, U. Shalev, J.N. Rawlins, I. Weiner, J. Feldon, S. Totterdell, Neonatal nonhandling and in utero prenatal stress reduce the density of NADPH-diaphorase-reactive neurons in the fascia dentata and Ammon's horn of rats, J. Neurosci. 17 (1997) 5599-5609.]. The present study investigated whether such a decrease in NADPHd activity would be detectable at earlier ages. Therefore, the present study assessed the density of nitric oxide (NO) producing neurons in the fascia dentata and Ammon's horn in 28-, 54-, and 118-day-old nonhandled and handled male rats using NADPHd histochemistry and immunohistochemical localization of neuronal isoform of nitric oxide synthase (nNOS), a NADPHd. This showed that in these three age groups, the numbers of NADPHd positive neurons per unit area throughout the hippocampus of rats that received no handling during neonatal development did not differ significantly from those of rats that received regular daily handling. In addition, we found in the rats of 118 days of age that the areal density of nNOS immunopositive neurons in the hippocampus also did not differ significantly between nonhandled and handled rats. Nevertheless, in a parallel study, rats from the same experimental group receiving identical treatments showed the expected impairment of latent inhibition at 4 months of age [R. Weizman, J. Lehmann, S. Leschiner, I. Allmann, T. Stoehr, C. Heidbreder, A. Domeney, J. Feldon, M. Gavish, Long-lasting effect of early handling on the peripheral-type benzodiazepine receptor, Pharmacol. Biochem. Behav. in press.]. These results suggest that nonhandling of rats during the early neonatal period, that does result in impairment in latent inhibition, does not affect the numbers of NO producing neurons in the hippocampus in rats of young ages, including the age of observed impairment of latent inhibition.

Pigeon basal ganglia: insights into the neuroanatomy underlying telencephalic sensorimotor processes in birds.

This paper reviews the organization of the avian and mammalian striatum. The striatum receives input from virtually the entire rostrocaudal and mediolateral expanse of the cerebral cortex. The corticostriatal projections appear to be glutamatergic, forming excitatory synapses in the striatum. Another major projection to the avian striatum that also appears to be glutamatergic stems from a set of nuclei in the dorsal zone of the avian thalamus that are comparable to the mammalian intralaminar, mediodorsal, and midline nuclei. Furthermore, the striatum receives a massive projection from dopaminergic neurons of the ventral tegmental area and substantia nigra in the midbrain tegmentum. In return, the midbrain tegmentum receives a direct GABAergic/substance P-ergic/ dynorphinergic projection from the striatum, as well as an indirect one formed by GABAergic/substance P-ergic/ dynorphinergic and GABA-ergic/enkephalinergic striatal neurons projecting to the pallidum in the first step, and pallidal GABAergic/LANT6/parvalbumin neurons projecting to the midbrain tegmentum in the second step. In addition to its projection neurons, the striatum possesses GABAergic and cholinergic interneurons. One motor output pathway of the striatum runs via the pallidum and dorsal thalamic ventral tier nulei to the motor cortex. In addition to this pathway, birds possess a major descending pathway from the basal ganglia to the tectum via the GABAergic nucleus spiriformis lateralis in the pretectum. On hodological and topological grounds, similar nuclei, although not GABAergic, can be found in mammals. Finally, an other striatal motor output is formed by a sequential GABAergic pathway from the basal ganglia via the substantia nigra to the tectum. In conclusion, it appears that the organization of the avian and mammalian basal ganglia is similar rather than different.

Cellular localization of AMPA type glutamate receptor subunits in the basal ganglia of pigeons (Columba livia).

Corticostriatal and thalamostriatal projections utilize glutamate as a neurotransmitter in mammals and birds. The influence on striatum is mediated, in part, by ionotropic AMPA-type glutamate receptors, which are heteromers composed of GluR1-4 subunits. Although the cellular localization of AMPA-type subunits has been well characterized in mammalian basal ganglia, their localization in avian basal ganglia has not. We thus carried out light microscopic single- and double-label and electron microscopic single-label immunohistochemical studies of GluR1-4 distribution and cellular localization in pigeon basal ganglia. Single-label studies showed that the striatal neuropil is rich in GluR1, GluR2, and GluR2/3 immunolabeling, suggesting the localization of GluR1, GluR2 and/or GluR3 to the dendrites and spines of striatal projection neurons. Double-label studies and perikaryal size distribution determined from single-label material indicated that about 25% of enkephalinergic and 25% of substance P-containing striatal projection neuron perikarya contained GluR1, whereas GluR2 was present in about 75% of enkephalinergic neurons and all substance-P -containing neurons. The perikaryal size distribution for GluR2 compared to GluR2/3 suggested that enkephalinergic neurons might more commonly contain GluR3 than do substance P neurons. Parvalbuminergic and calretininergic striatal interneurons were rich in GluR1 and GluR4, a few cholinergic striatal interneurons possessed GluR2, but somatostatinergic striatal interneurons were devoid of all subunits. The projection neurons of globus pallidus all possessed GluR1, GluR2, GluR2/3 and GluR4 immunolabeling. Ultrastructural analysis of striatum revealed that GluR1 was preferentially localized to dendritic spines, whereas GluR2/3 was found in spines, dendrites, and perikarya. GluR2/3-rich spines were generally larger than GluR1 spines and more frequently possessed perforated post-synaptic densities. These results show that the diverse basal ganglia neuron types each display different combinations of AMPA subunit localization that shape their responses to excitatory input. For striatal projection neurons and parvalbuminergic interneurons, the combinations resemble those for the corresponding cell types in mammals, and thus their AMPA responses to glutamate are likely to be similar.

Avian homologues of mammalian intralaminar, mediodorsal and midline thalamic nuclei: immunohistochemical and hodological evidence.

This paper presents and reviews data suggesting that the dorsal thalamic zone (abbreviated DTZ) in birds is homologous to the intralaminar, midline, and mediodorsal thalamic nuclear complex (abbreviated IMMC) in mammals. The DTZ is located dorsomedially in the diencephalon of birds and consists of several subnuclei: nucleus dorsomedialis anterior thalami (DMA), nucleus dorsomedialis posterior thalami (DMP), nucleus dorsolateralis anterior thalami, pars medialis (DLM), nucleus dorsointermedius posterior thalami (DIP), nucleus dorsolateralis posterior thalami (DLP), and nucleus subhabenularis lateralis (SHL). Our immunohistochemical studies show that: (1) SHL and medial and dorsal parts of DMA and DMP are relatively rich in GABAergic, enkephalin-containing, substance P-containing, and cholinergic fibers; (2) lateral parts of DMA and DMP are relatively poor in these neurotransmitters; and (3) DIP, DLP, and DLM are moderately rich in cholinergic and substance P-containing fibers. Our retrograde pathway tracing studies indicate that the DIP and DLP in the more lateral parts of DTZ project to somatic striatum, while the DMA, DMP, and SHL located more medially in the DTZ project to visceral/limbic striatum. Our anterograde tracing studies indicate that DIP receives afferents from the dorsal pallidum, whereas DMA and DMP appear to receive afferents from both the ventral striatum and ventral pallidum. Diverse prior studies have shown that in general medial and lateral components of DTZ are connected with visceral/ limbic and somatic brain regions, respectively. These characteristics indicate that: (1) SHL and medial and dorsal parts of DMA and DMP are comparable to mammalian midline thalamic nuclei, including the medial components of the intralaminar nuclei; (2) lateral parts of DMA and DMP are comparable to the mediodorsal nucleus in mammals; (3) DIP is comparable to the parafascicular nucleus in mammals; and (4) DLM and DLP are comparable to the laterally located intralaminar nuclei in mammals. The comparability of avian DTZ and mammalian IMMC suggests that they evolved from thalamic precursor nuclei present in the common reptilian ancestors and that they may perform similar roles in the movement control function of the basal ganglia.