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Coexistence of parvalbumin and GABA in nonpyramidal neurons of the rat entorhinal cortex.

The possible coexistence of the calcium-binding protein, parvalbumin, with the major inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), and its synthesizing enzyme, glutamate decarboxylase (GAD), was studied in nonpyramidal cells of the rat medial and lateral entorhinal cortex. The material was analyzed by two different methods, the first of which was a mirror techniques where the possible coexistence of two different antigens was analyzed from cells cut in half at the surface of the adjacent section. The other method consisted of analyzing double immunofluorescent-stained sections with a confocal microscope. The colocalization analysis revealed that all parvalbumin-immunoreactive neurons (mirror technique n = 688 and confocal microscopy n = 644) in all layers of the medial and lateral entorhinal cortex were also immunopositive for GABA or GAD. Parvalbumin-cells made up 52% of the GABA cells in most of the layers in the medial and lateral entorhinal cortex. In layer III of the entorhinal cortex, the proportion was about 40%. Thus, parvalbumin-containing neurons in the entorhinal cortex represent a large GABAergic cell population, which is likely to play an important role in controlling both the input and the output of the entorhinal cortex.

Animals↗

The hierarchical development of monkey visual cortical regions as revealed by the maturation of parvalbumin-immunoreactive neurons.

The prefrontal cortex is known to be involved in behavioral paradigms requiring decisions based on short-term working memory, and visually related areas of prefrontal cortex represent the final point in a proposed hierarchical sequence of visual signal processing that begins in primary visual cortex. This study asks if the development of at least certain aspects of the circuitry of each region involved in this hierarchy proceeds in a sequential fashion from primary to higher-order areas. The timing and patterns of expression of immunoreactivity for the calcium-binding protein parvalbumin were examined in areas V1, V2, TE, 7a, and 46 in two series of macaque monkeys ranging in age from embryonic day 132 to adult. The number and laminar distribution of parvalbumin-labeled neurons reached adult levels first in area V1 (primary visual cortex), followed by the adjacent visual association area V2, and then by the higher-order regions of the inferior temporal (TE), posterior parietal (7a) and prefrontal (46) cortices. The appearance of parvalbumin immunoreactivity in the axons of the two major classes of local circuit neurons that express this protein, basket and chandelier cells, followed a similar regional pattern. Furthermore, striking differences were present between these two neuronal populations in the laminar pattern and time course of parvalbumin labeling of their axons. These findings demonstrate that at least some aspects of the intrinsic circuitry of the neocortex mature in accordance with a functional hierarchy of cortical regions. In addition, they illustrate the complexity of cortical development in terms of the different timing of expression of even a single protein in different compartments within single neurons, in different cell types, in different laminae within a region, and across different cortical regions.

Animals↗

Sodium and potassium binding to parvalbumins measured by means of intrinsic protein fluorescence.

The binding of Na+ and K+ to whiting parvalbumin (pI 4.4) and pike parvalbumins (pI 4.2 and 5.0) results in a shift of the tryptophan fluorescence spectrum towards shorter wavelengths by 2-4 nm for the whiting protein and in a rise of the tyrosine and phenylalanine fluorescence quantum yield for the pike proteins. The effective binding constants of Na+ and K+ to parvalbumins are within the range of 10 M-1 to 100 M-1. Physiological concentrations of Na+ and K+ lower the affinity of whiting parvalbumin for Ca2+ and Mg2+ by almost an order of magnitude.

Animals↗

Age-related changes in parvalbumin- and GABA-immunoreactive cells in the rat septum.

The calcium binding protein parvalbumin is present in GABAergic neurons of the medial septum-diagonal band of Broca (MS-DBB) region that project to the hippocampal formation. We examined the distribution pattern, the number, and the morphological features of the parvalbumin-containing cells (parv+) in the MS-DBB region of 2- to 3-, 8- to 9-, 15- to 16-, and 26- to 27-month-old Sprague-Dawley rats. A significant reduction in the number of parv+ cells was observed as a function of age. The mean somal area of the parv+ cells was significantly reduced in the 26- to 27-month-old rats. A significant reduction in the number of parv+ cells was also observed in the 26- to 27-month-old rats in the cingulate cortex, but not in the striatum or the hippocampal formation. No significant age-related changes were observed in the number of the GABA-immunoreactive cells in the MS-DBB region nor in the cingulate cortex. In conclusion, there is an age-related decrease in the number of parv+ cells, with no change in the number of GABA-immunoreactive cells in the MS-DBB region of the rat. Because GABA and parvalbumin are colocalized in the MS-DBB neurons, the results suggest that the level of parvalbumin is decreased, but that the cells are not lost.

Aging↗

Interaction of cupric ion with parvalbumin.

Cod parvalbumin, a calcium-binding protein, possesses a specific Zn2+ (or Cu2+) binding site per molecule. This work employed fluorescence energy transfer techniques to measure the distance between the Zn2+ (Cu2+) site and the stronger Ca(2+)-binding site in parvalbumin. Specifically, the distance between Tb3+ bound at the Ca2+ site and Co2+ bound to the Zn2+ (Cu2+) binding site was 10.3 +/- 0.9 A. Lastly, the effects of Cu2+ on the physico-chemical properties of parvalbumin were studied by measuring the accessibility of protein thiol groups to 5,5'-dithio bis(2-nitrobenzoic acid) and by its affinity for the fluorescent probe 4,4'-bis[1-(phenylamino)-8-naphthalene sulfonic acid] dipotassium salt. The thiol group accessibility decreased and the affinity to the fluorescent probe increased upon complexation of Cu2+ to the protein. It appears that the binding of Cu2+ converts parvalbumin to an apo-like state.

Animals↗

Demonstration of neurofibrillary tangles in parvalbumin-immunoreactive interneurones in the cerebral cortex of Alzheimer-type dementia brain.

The pathological changes occurring in a population of parvalbumin-like immunoreactive cerebral cortical interneurones in postmortem Alzheimer-type dementia brain tissue were determined by double staining cerebral cortical sections for parvalbumin, tau and/or calpain-like immunoreactivities. By counting the number of double immunostained cells it was determined that 4% parvalbumin and 25% of calpain-like immunoreactive cells showed evidence of tau-like immunoreactivity, indicative of neurofibrillary tangles. These results suggest that cerebral cortical parvalbumin-like immunoreactive neurones are relatively resistant to the pathological changes underlying the formation of neurofibrillary tangles associated with Alzheimer-type dementia.

Alzheimer Disease↗

Parvalbumin-immunoreactive dystrophic neurites and aberrant sprouts in the cerebral cortex of patients with Alzheimer's disease.

Parvalbumin-immunoreactive dystrophic neurites and aberrant terminal sprouts associated with senile plaques, together with preserved density of parvalbumin-immunoreactive neurons, were found in the cerebral cortex of two patients with Alzheimer's disease, one of them familiar, in which a biopsy of the left frontal lobe was carried out for diagnostic and counselling purposes. These findings suggest that, although parvalbumin-immunoreactive cells are relatively resistant to degeneration in Alzheimer's disease, parvalbumin-immunoreactive neuronal processes can degenerate in some cases of Alzheimer's disease.

Aged↗

Changes in parvalbumin-immunoreactive neurons in the rat hippocampus following a kainic acid lesion.

Changes in a sub-population of hippocampal non-pyramidal neurons following a unilateral lesion with kainic acid were examined using an antibody raised against the Ca-binding protein parvalbumin. A loss of 71-97% of the parvalbumin-immunoreactive neurons occurred at the three post-lesion times studied (1, 2 and 4 weeks) in all areas of the ipsilateral hippocampus, but no such loss was observed in the dentate gyrus. Resistant parvalbumin-immunoreactive neurons occurred principally in stratum pyramidale and displayed altered morphology from the normal with swollen dendrites and dendritic varicosities. The contralateral hippocampus exhibited losses of parvalbumin-immunoreactive cells, but this was restricted to stratum oriens of CA1. This data demonstrates the loss of a specific and important population of non-pyramidal neurons which might be responsible for the chronic loss of functional inhibition seen in this animal model of temporal lobe epilepsy.

Animals↗

Parvalbumin as an anatomical marker for discrete subregions of the ambiguus complex in the rat.

The topography of parvalbumin-immunoreactive neurons within the ventrolateral medulla of rats was investigated. Parvalbumin is a member of the 'EF-hand' family of Ca-binding proteins and is present in certain cell types within the central nervous system (fast-firing neurons with high metabolic rates). Parvalbumin-immunoreactive neurons were located in discrete rostrocaudal divisions of the ambiguus complex corresponding to regions containing respiratory-related neurons. Based on the location of physiologically characterized respiratory-related neurons reported in the literature, parvalbumin immunoreactivity does not appear to distinguish inspiratory- from expiratory-related neurons.

Animals↗

Parvalbumins. Distribution and physical state inside the muscle cell.

Single skinned muscle fibres (frog) have been submitted to double Ouchterlony immunodiffusion assays with antibodies directed against the two species of frog parvalbumins. The antigenic material which diffuses out of each fibre contains the two parvalbumins. Their presence in each cell is thus demonstrated. The amount of parvalbumins having diffused out of the fibre has been quantified. It corresponds to the parvalbumin content of the cell. This implies that these proteins are freely soluble in the muscle sarcoplasm.

Animals↗

Calbindin-D-28k-like immunoreactive structures in the olfactory bulb and anterior olfactory nucleus of the human adult: distribution and cell typology--partial complementarity with parvalbumin.

Calbindin-D-28k and parvalbumin are calcium-binding proteins. The laminar distribution and morphological features of calbindin-D-28k-like immunoreactive structures were studied in 60-microns-thick sections of the human olfactory bulb. Except for the olfactory nerve layer, immunoreactive neurons were present in all layers of the olfactory bulb. They reached highest densities in the external plexiform layer and internal granule cell layer. Considerable numbers of calbindin-like nerve cells were also found in the olfactory tract and in distal portions of the anterior olfactory nucleus. When comparing the distribution of calbindin-positive structures to that of parvalbumin-positive ones a partially complementary distribution pattern was found. Calbindin-like immunoreactive portions of the anterior olfactory nucleus and olfactory tract were mirrored by immunonegative areas in adjacent sections stained for parvalbumin. Using the combined pigment-Nissl procedure we observed the presence of lipofuscin deposits in nearly 80% of all the calbindin-immunoreactive neurons analysed. Moreover, analysis of their lipofuscin deposits rendered the further differentiation of morphologically similar neuronal subpopulations possible. In contrast, all parvalbumin-like immunoreactive neurons remained free of lipofuscin granules.

Adult↗

Presence of calbindin and lack of parvalbumin in progesterone receptor-containing neurons of the monkey mediobasal hypothalamus.

All of the progesterone receptor-containing cells of the monkey hypothalamus are GABAergic. The aim of this study was to further characterize these GABAergic progesterone receptor-containing neurons based on their calbindin or parvalbumin content. These calcium-binding proteins are characteristic markers of different populations of GABAergic neurons in the central nervous system. Double-immunolabeling for progesterone receptor and either calbindin or parvalbumin was performed on hypothalamic Vibratome sections of estrogen primed African green monkeys (Cercopithecus aethiops). Progesterone receptor-containing calbindin-immunoreactive neurons were observed in the ventromedial and periventricular areas of the hypothalamus. Forty-one per cent of the progesterone receptor-containing cells in this area were calbindin immunopositive. No double-immunolabeled neurons could be detected in the infundibular (arcuate) nucleus. In tissue double-immunolabeled for progesterone receptor and parvalbumin, none of the progesterone receptor-containing neurons exhibited immunoreactivity for parvalbumin. Electron microscopic double-immunostaining for progesterone receptor and calbindin confirmed the light microscopic results. Furthermore, a large number of asymmetric synaptic contacts were observed on the calbindin-immunoreactive neurons. These observations demonstrate that progesterone receptor-containing cells in the monkey mediobasal hypothalamus consist of at least two different types of GABA neurons, and indicate that progesterone receptor-containing calbindin cells may be postsynaptic targets of excitatory fibers.

Animals↗

Neocortical, hippocampal and septal parvalbumin- and somatostatin-containing neurons in young and aged rats: correlation with passive avoidance and water maze performance.

Aged (26-month-old) rats were impaired compared with young (three-month-old) rats in passive avoidance and water maze tasks. In order to study whether changes in inhibitory circuits are involved in these age-related cognitive impairments, the number of two different subpopulations of GABAergic neurons, i.e. somatostatin- and parvalbumin-containing neurons, were counted in the hippocampal formation, septum and neocortex. We found that the number of parvalbumin-containing neurons was decreased in the entorhinal, somatosensory and motor cortex as well as in the medial septum and vertical limb of the diagonal band of Broca, but not in the hippocampus of aged rats. Somatostatin-containing neurons were affected in the somatosensory and motor cortex, and in the dorsolateral septum, but not in the hippocampus or in the entorhinal cortex. The decreased number of parvalbumin-containing neurons in the entorhinal cortex of the aged rats correlated with their performance deficits in passive avoidance and spatial learning. We propose that impaired functioning of the entorhinal cortex parvalbumin-containing inhibitory neurons may, to some extent, be responsible for the learning and memory defects found in aged rats.

Aging↗

Subpopulations of GABAergic neurons in laminae I-III of rat spinal dorsal horn defined by coexistence with classical transmitters, peptides, nitric oxide synthase or parvalbumin.

GABAergic neurons in laminae I-III of the spinal dorsal horn may contain one or more of the following compounds: glycine, acetylcholine, neuropeptide Y, enkephalin, nitric oxide synthase or parvalbumin. Although the pattern of co-localization of some of these compounds is understood, it is not known which types of GABAergic neurons contain parvalbumin, or whether nitric oxide synthase coexists with peptides, acetylcholine or parvalbumin in any of these neurons, and in this study we have used immunocytochemistry and enzyme histochemistry to resolve these issues. Parvalbumin-immunoreactivity was restricted to those GABA-immunoreactive neurons that also showed glycine-immunoreactivity and was not co-localized with neuropeptide Y-immunoreactivity or NADPH diaphorase activity. By combining NADPH diaphorase histochemistry with immunocytochemistry with an antiserum to nitric oxide synthase, we were able to show that NADPH diaphorase activity was a reliable marker for nitric oxide synthase in the spinal cord. Neurons that possess GABA- but not glycine-immunoreactivity may contain neuropeptide Y, enkephalin, acetylcholine or NADPH diaphorase, and all of the cholinergic neurons appear to contain NADPH diaphorase. By combining immunofluorescent detection of neuropeptide Y or enkephalin with NADPH diaphorase histochemistry, we showed that peptide-immunoreactivity did not coexist with NADPH diaphorase. This suggests that neither of these peptides coexists with nitric oxide synthase or with acetylcholine in neurons in the superficial dorsal horn. Several phenotypically distinct groups of GABA-immunoreactive neuron can therefore be identified in laminae I-III of the dorsal horn, and these may represent different functional types of inhibitory neuron.

Acetylcholine↗

Chronic intrastriatal quinolinic acid produces reversible changes in perikaryal calbindin and parvalbumin immunoreactivity.

We recently reported the use of a chronic dialytic delivery system for intrastriatal administration of quinolinic acid in the rat. This system produces neurodegeneration with some characteristics similar to post mortem brain tissue from Huntington's disease patients, including reduced cytochrome oxidase staining, a decreased number of Nissl-stained neurons, and relative sparing of striatal NADPH-diaphorase containing neurons. The present findings show that chronic dialytic delivery of quinolinic acid also produces a Huntington's disease-like pattern of reduced calbindin and parvalbumin perikaryal immunoreactivity that is reversed in rats allowed four to eight weeks' recovery after cessation of quinolinic acid. Furthermore, cytochrome oxidase staining and the number of Nissl-stained cells were unchanged in the region of transient calbindin and parvalbumin immunoreactive perikaryal staining alterations. These results suggest that changes in calbindin and parvalbumin perikaryal immunoreactivity provide a relatively sensitive measure of quinolinic acid induced neurotoxicity. The reversible nature of reduced perikaryal immunoreactivity suggests a premorbid state of neurotoxicity, possibly marked by cellular redistribution of calbindin and parvalbumin.

Animals↗

The chemical heterogeneity of cortical interneurons: nitric oxide synthase vs. calbindin and parvalbumin immunoreactivity in the rat.

Neurons that contain nitric oxide synthase (NOS) type I and the calcium binding proteins calbindin D28k or parvalbumin were simultaneously visualized by means of double immunohistofluorescence in the cerebral cortex of Wistar and Sprague-Dawley rats. All the three immunoreactive cell populations were primarily represented by nonpyramidal neurons. NOS-immunoreactive cells were less numerous than the calbindin- or parvalbumin-immunoreactive ones, and were intermingled with the neurons containing these calcium binding proteins. NOS-immunoreactive cells were separate from the parvalbumin-immunoreactive ones, whereas a minor proportion of them was found to be colocalized with calbindin. The cortical neurons in which NOS and calbindin coexisted were more numerous in the Sprague-Dawley than in the Wistar rats, and displayed an anteroposterior gradient of density, with the highest concentration in the medial prefrontal, frontal, and cingulate cortices. Double NOS-calbindin-immunoreactive neurons prevailed in the deep cortical layers and they were relatively numerous in the cingulate cortex. The present data indicate a selectivity in the expression of NOS vs. calbindin and parvalbumin in cortical cells, and further support the chemical heterogeneity of GABAergic interneurons in the cerebral cortex.

Animals↗

The genes for the highly homologous Ca(2+)-binding proteins oncomodulin and parvalbumin are not linked in the human genome.

The chromosomal loci of the human parvalbumin and oncomodulin single-copy genes that encode structurally and evolutionarily closely related Ca(2+)-binding proteins were determined by somatic cell hybrid analysis. Southern blot analysis of genomic DNA from 25 human-hamster somatic cell hybrids showed that the human gene for oncomodulin resides on chromosome 7. Analysis of human-mouse hybrids selectively retaining human chromosome 7 or a portion of it allowed specific assignment of the gene locus to the p11-p13 region of chromosome 7 known to be mutated or deleted in patients with the Greig cephalopolysyndactyly syndrome. By gene dosage analysis on Southern blots, we showed that the gene for human parvalbumin maps distally to the cat eye syndrome marker D22S9 on chromosome 22q. Using somatic cell hybrids containing parts of human chromosome 22, the parvalbumin gene was sublocalized to the region 22q12-q13.1. This region contains a linkage group that maps to mouse chromosome 15, region E, and includes the SIS, ARSA, and DIA 1 genes. Our findings are consistent with the recent localization of the mouse parvalbumin gene to this region by two independent methods (C. H. Zühlke et al., 1989, Genet. Res. 54:37-43; S. Adolph et al., 1989, Cytogenet. Cell Genet. 52:177-179).

Animals↗

Effect of Brn-3a deficiency on parvalbumin-immunoreactive primary sensory neurons in the dorsal root ganglion.

Immunohistochemistry for parvalbumin, a marker for primary proprioceptors, was performed on the dorsal root ganglion (DRG) of wildtype and knockout mice for Brn-3a at postnatal day 0 and embryonic day 18.5. The DRG contained many parvalbumin-immunoreactive (ir) neurons in wildtype (5.4%) and knockout mice (5.6%). Cell size analysis demonstrated that such neurons were mostly medium-sized to large in these mice. Therefore, it is unlikely that the survival of proprioceptors is dependent upon Brn-3a in the DRG. In the dorsal column and gray matter of the spinal cord of knockout mice, however, parvalbumin-ir nerve fibers were sparse compared to wildtype mice. The number of parvalbumin-ir varicosities around motoneurons decreased in the mutant. Thus, our data suggest that Brn-3a may play an important role in the central projection and terminal formation of DRG proprioceptors in the spinal cord.

Afferent Pathways↗