Calretinin-immunoreactive terminals make synapses on calbindin D28k-immunoreactive neurons in the lateral nucleus of the human amygdala.

A double-labeling immunohistochemical procedure and correlated light and electron microscopy were used to examine if calretinin-immunoreactive terminals make synapses on calbindin D28k-positive cells. In the lateral nucleus of the human amygdala, calretinin terminals make symmetric-like synapses on the somata and proximal dendrites of calbindin D28k-labeled cells. Our data provide the first evidence that neurons which contain two different calcium-binding proteins form synaptic contacts with each other in the human amygdala.

Calbindin-D28K-immunoreactive cells and fibres in the human amygdaloid complex.

The distribution of calbindin-D28k-immunoreactive cells and fibres in five human amygdalae was analysed from sections that had been stained immunohistochemically with a monoclonal antibody raised against calbindin-D28k. The highest density of calbindin-D28k-positive neurons were found in the anterior cortical, medial, posterior cortical and accessory basal nuclei, in the parvicellular division of the basal nucleus and in the amygdalohippocampal area. The lowest densities of immunopositive neurons were found in the paralaminar nucleus, in the periamygdaloid cortex (PAC1 and PACo) and in some of the intercalated nuclei. The deep nuclei (lateral, basal and accessory basal nuclei) contained a high density of calbindin-D28k-immunoreactive fibres and terminals. The cortical nuclei and the central nucleus were characterized by intense neuropil labelling. Morphologically, a large majority of the calbindin-D28k-immunoreactive neurons were aspiny or sparsely spiny and resembled inhibitory local circuit neurons. A small population of lightly-stained, pyramidal-shaped neurons was also observed. In most of the amygdaloid nuclei, calbindin-D28k-immunoreactive fibres travelled close to each other and formed bundles, which suggests that some of the immunostained neurons were double-bouquet cells. In the paralaminar nucleus, the calbindin-D28k-immunoreactive axons formed tortuous plexus (100-200 microns in diameter) that surrounded several unstained somata. This study provides baseline information on the morphology and distribution of calcium-binding protein-containing inhibitory cells and fibres immunoreactive for calbindin-D28k in the human amygdaloid complex. This information can be used in future studies on the pathogenesis of diseases known to damage the amygdala, such as Alzheimer's disease and temporal lobe epilepsy.

Parvalbumin-immunoreactive neurons make inhibitory synapses on pyramidal cells in the human amygdala: a light and electron microscopic study.

In the present study we investigate the inhibitory circuitries that regulate the neuronal activity in the lateral and basal nuclei, which are the main sensory input regions of the amygdala. Axon terminals immunoreactive for parvalbumin, a calcium-binding protein known to colocalize with GABA, were examined in these regions with electron microscopy, and their postsynaptic targets were identified and characterized. In the lateral nucleus, parvalbumin-immunoreactive (PV-ir) axons formed terminal rows which made symmetric synaptic contacts on the axon initial segments of the pyramidal cells. In the basal nucleus, pericellular baskets of PV-ir fibers established symmetric synapses on pyramidal cell somata and proximal dendrites. Our data suggest that PV-ir neurons play a crucial inhibitory role in the control of pyramidal cell activity in the human amygdala.

Calretinin-immunoreactive cells and fibers in the human amygdaloid complex.

Calretinin is a calcium-binding protein that colocalizes with GABA in the cerebral cortex and hippocampus of the rat and the monkey. In the present study, we investigated the distribution of calretinin-immunoreactive cells and fibers in the human amygdaloid complex. A conspicuous feature was the high density of calretinin neurons in the human amygdala. The highest densities of the calretinin-immunoreactive neurons were observed in the anterior cortical nucleus, accessory basal nucleus, amygdalohippocampal area, and in the nucleus of the lateral olfactory tract. The paralaminar nucleus, central nucleus, medial nucleus, and the periamygdaloid cortex contained the lowest densities of calretinin neurons. In most of the amygdaloid areas, the calretinin cells had the appearance of aspiny or sparsely spiny local circuit neurons. However, in the amygdalohippocampal area, we found also densely spined dendrites. The cortical areas and the central nucleus were characterized by intense neuropil labeling, while the deep nuclei contained a high density of calretinin-immunoreactive fibers and terminals. Calretinin immunoreactivity was also found in the intra-amygdaloid fiber bundles, stria terminalis, and in the ventral amygdalofugal pathway. This suggests that in addition to the local circuit neurons, calretinin immunoreactivity is also located in neurons that connect the amygdaloid complex with the other brain areas. The distribution and morphological characteristics of calretinin-immunoreactive neurons differed from those of another calcium-binding protein, parvalbumin, in the human amygdala (Sorvari et al. [1995] J. Comp. Neurol. 360:185-212). This suggests that these two calcium-binding proteins are located in different populations of neurons.

Distribution of parvalbumin-immunoreactive cells and fibers in the human amygdaloid complex.

The calcium-binding protein, parvalbumin, was localized immunohistochemically in the human amygdaloid complex. Neuronal cell bodies and fibers that are immunoreactive to parvalbumin were observed in most of the amygdaloid nuclei and cortical areas. Three types of immunoreactive aspiny neurons, ranging from small spherical cells (type 1) to large multipolar cells (type 2) and fusiform cells (type 3), were observed. The densities of the types of neurons that were parvalbumin-immunoreactive varied in the different regions of the amygdala. The highest densities of parvalbumin-immunoreactive neurons were observed in the lateral nucleus, in the magnocellular and intermediate divisions of the basal nucleus, in the magnocellular division of the accessory basal nucleus and in the amygdalohippocampal area. The regions containing the lowest density of parvalbumin-immunoreactive cells were the paralaminar nucleus, the parvicellular division of the basal nucleus, the central nucleus, the medial nucleus and the anterior cortical nucleus. In general, the distribution of immunoreactive fibers and terminals paralleled that of immunoreactive cells. Parvalbumin-immunoreactive varicose fibers formed basket-like plexi and cartridges around the unstained neurons, which suggests that parvalbumin is located in GABAergic basket cells and chandelier cells, respectively. The distribution of parvalbumin-immunoreactive profiles in the human amygdaloid complex was similar to, rather than different from that previously reported in the monkey amygdala (Pitkänen and Amaral [1993] J. Comp. Neurol. 331:14-36). This study provides baseline information about the organization of GABAergic inhibitory circuitries in the human amygdaloid complex.

Alzheimer pathology of patients carrying apolipoprotein E epsilon 4 allele.

A recent report suggested that brains of Alzheimer patients homozygous for APOE epsilon 4 show increased amyloid pathology compared to APOE epsilon 3 homozygotes. We studied APOE allele frequencies in 73 AD patients and 38 controls. We also investigated relation of APOE genotypes to beta/A4 immunopositive plaques, cerebrovascular beta/A4 deposition, neurons expressing paired helical filaments (PHFs), and synaptophysin-like immunopositivity in 22 neuropathologically verified AD patients. We also correlated APOE genotypes of definite AD patients to beta/A4 immunoreactivity in dermal vessel walls detected in lifetime skin biopsy samples. APOE allele epsilon 4 frequency was increased in AD compared to nondemented controls (0.37 vs. 0.11; p = 0.006). The number of beta/A4 immunoreactive plaques, PHFs-containing neurons, the degree of cerebrovascular beta/A4 deposition or synaptophysin-like immunoreactivity did not differ significantly in AD patients with or without epsilon 4. beta/A4 deposition in dermal vessel walls was more frequent in definite AD patients with epsilon 4 (43%) than in patients without epsilon 4 (22%). However, the difference did not reach the statistical significance.

Loss of synaptophysin-like immunoreactivity in the hippocampal formation is an early phenomenon in Alzheimer's disease.

We studied a synatophysin-like immunoreactivity in the hippocampal formation of patients with definite Alzheimer's disease, multi-infarct dementia, patients with no evidence of clinical dementia with neuropathological findings fulfilling the criteria of possible Alzheimer's disease, and age-matched nondemented controls. Possible Alzheimer's disease cases were of special interest because they were considered to represent early Alzheimer's disease. We also studied the spatial relationship of synaptophysin-like immunopositivity with amyloid-beta-protein immunopositive senile plaques and anti-paired helical filament immunopositive degenerating neurons locally as well as considering the intrinsic circuits in the hippocampal formation. The synaptophysin-like immunoreactivity was decreased in the hippocampus and the entorhinal cortex in patients with definite and possible Alzheimer's disease but not in multi-infarct dementia patients compared to controls. Equal loss of synapses in possible and definite Alzheimer's disease patients supports the hypothesis that synaptic loss is an early phenomenon in Alzheimer's disease. Unchanged synaptophysin-like immunopositivity in patients with multi-infarct dementia suggests that the loss of synapses is centrally involved in the pathogenesis of Alzheimer's disease and not dementia per se. There was no spatial correlation between loss of synapses and amyloid-beta-protein positive senile plaques. Moreover, we could not find a strict spatial relationship between senile plaques and degenerating neurons. Our results do not support the amyloid cascade hypothesis of Alzheimer's disease that local accumulation of amyloid-beta-protein leads to the loss of synapses.