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PubMed2003

Diffusional mobility of parvalbumin in spiny dendrites of cerebellar Purkinje neurons quantified by fluorescence recovery after photobleaching.

Ca(2+)-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca(2+) dynamics. Especially in thin extensions of nerve cells, Ca(2+) binding and buffered diffusion of Ca(2+) by CaBPs is assumed to effectively control the spatio-temporal extend of Ca(2+) signals. However, no quantitative data about the mobility of specific CaBPs in the neuronal cytosol are available. We quantified the diffusion of the endogenous CaPB parvalbumin (PV) in spiny dendrites of cerebellar Purkinje neurons with two-photon fluorescence recovery after photobleaching. Fluorescently labeled PV diffused readily between spines and dendrites with a median time constant of 49 ms (37-61 ms, interquartile range). Based on published data on spine geometry, this value corresponds to an apparent diffusion coefficient of 43 microm(2) s(-1) (34-56 microm(2) s(-1)). The absence of large or immobile binding partners for PV was confirmed in PV null-mutant mice. Our data validate the common but so far unproven assumption that PV is highly mobile in neurons and will facilitate simulations of neuronal Ca(2+) buffering. Our experimental approach represents a versatile tool for quantifying the mobility of proteins in neuronal dendrites.

Animals↗
PubMed1983

Anomalous x-ray scattering from terbium-labeled parvalbumin in solution.

We have used anomalous small-angle x-ray scattering as a structural probe for solutions of rabbit parvalbumin labeled with terbium. This technique makes use of the large changes in the terbium scattering factor that occur when the x-ray energy is tuned around an L3 absorption edge of this heavy-atom label. These changes in scattering result in changes in the small-angle scattering curve of the labeled protein as a whole, which can then be analyzed to derive structural information concerning the distribution of labels in the protein. Based on a Gaussian model for the protein electron density, the mean distance from the terbiums to the protein center of mass is determined to be 13.2 A and is consistent with crystallographic results. Our results demonstrate the usefulness of terbium as an anomalous scattering label and provide criteria to help establish anomalous scattering as a reliable structural technique for proteins in solution.

Animals↗
PubMed1999

Intracellular EDTA mimics parvalbumin in the promotion of skeletal muscle relaxation.

Parvalbumin (PA) is an intracellular Ca2+-binding protein found in some muscle and nerves. Its ability to bind Ca2+ and facilitate skeletal muscle relaxation is limited by its Mg2+ off-rate. EDTA serves as an "artificial" PA in that it exhibited similar rate constants for Mg2+ (3 s-1) and Ca2+ (0.7 s-1) dissociation at 10 degrees C. When introduced into frog skeletal muscle, EDTA increased the relaxation rate by approximately 2.7-fold, and with increasing tetanus duration, EDTA lost its ability to contribute to relaxation (and Ca2+ sequestration) at its Mg2+ off-rate. Intracellular EDTA recovered its ability to contribute to muscle relaxation and Ca2+ sequestration at its Ca2+ off-rate. Like PA, EDTA's contribution to muscle relaxation and Ca2+ sequestration was more clearly observed when the SR Ca-ATPase was inhibited. Introduction of EDTA into rat soleus muscle, which has low [PA], increased the relaxation rate in a manner that was analogous to the way in which PA facilitates relaxation of frog skeletal muscle. Thus intracellular EDTA serves as an effective mimic of PA, and its use should aid in our understanding of PA's function in muscle and nerve.

Animals↗
PubMed1999

Hydration-coupled dynamics in proteins studied by neutron scattering and NMR: the case of the typical EF-hand calcium-binding parvalbumin.

The influence of hydration on the internal dynamics of a typical EF-hand calciprotein, parvalbumin, was investigated by incoherent quasi-elastic neutron scattering (IQNS) and solid-state 13C-NMR spectroscopy using the powdered protein at different hydration levels. Both approaches establish an increase in protein dynamics upon progressive hydration above a threshold that only corresponds to partial coverage of the protein surface by the water molecules. Selective motions are apparent by NMR in the 10-ns time scale at the level of the polar lysyl side chains (externally located), as well as of more internally located side chains (from Ala and Ile), whereas IQNS monitors diffusive motions of hydrogen atoms in the protein at time scales up to 20 ps. Hydration-induced dynamics at the level of the abundant lysyl residues mainly involve the ammonium extremity of the side chain, as shown by NMR. The combined results suggest that peripheral water-protein interactions influence the protein dynamics in a global manner. There is a progressive induction of mobility at increasing hydration from the periphery toward the protein interior. This study gives a microscopic view of the structural and dynamic events following the hydration of a globular protein.

Alanine↗
PubMed2000

Striatal interneurons expressing calretinin, parvalbumin or NADPH-diaphorase: a comparative study in the rat, monkey and human.

The present study is aimed at evaluating the relative number and comparing the pattern of distribution of interneurons containing calretinin (CR), parvalbumin (PV) or NADPH-d in the striatum of rats, squirrel monkeys and humans. A series of adjacent coronal sections taken at three comparable rostrocaudal levels in the three species were treated to reveal the three neuronal markers and the density of each type of chemospecific interneurons was analyzed with a computerized image analysis system. In primates, the most abundant interneurons were those expressing CR. The ratio of CR+/PV+ neurons was approximately 2-3:1 compared to a ratio of 3-4:1 for CR+/NADPH-d+ neurons. In contrast, the most frequently encountered interneurons in the rat striatum were those expressing PV. In rodents, all three interneurons were more abundant rostrally than caudally, but CR+ neurons displayed a particularly striking rostrocaudal decreasing gradient. In monkeys and humans, the three striatal interneurons were distributed rather uniformly rostrocaudally, but CR+ and PV+ interneurons were significantly more numerous in the caudate nucleus than in the putamen in humans. In monkeys, only PV+ neurons were more abundant in the caudate nucleus than in putamen. Overall, the density of the three striatal interneurons was much higher in monkeys than in rats and humans. These results reveal important species differences in respect to the relative density and pattern of distribution of striatal interneurons. These findings should be taken into account when evaluating the effect of neurodegenerative processes on cell densities in the human striatum or when studying animal models of the such diseases.

Animals↗
PubMed2000

GFR alpha-1 is expressed in parvalbumin GABAergic neurons in the hippocampus.

Glial cell line derived neurotrophic factor (GDNF) is a potent survival factor for several types of neurons. GDNF binds with high affinity to GDNF-family receptor alpha-1 (GFR alpha-1). This receptor is expressed in different areas of the brain, including the hippocampus and dentate gyrus. By using in situ hybridization and immunohistochemistry, we found that 19% to 37% of glutamic acid decarboxylase (GAD) expressing neurons co-expressed GFR alpha-1 in the hippocampus. GFR alpha-1/GAD co-expression was found mainly in the stratum (s) pyramidale (29-37%) and s. oriens (20-25%). Further characterization of GFR alpha-1 expressing interneurons, based on their calcium-binding protein immunoreactivity, demonstrated that many parvalbumin (PV) immunoreactive neurons express GFR alpha-1 in the s. pyramidale of CA1 (72%), CA2 (70%) and CA3 (70%) subfields of the hippocampus. GFR alpha-1/PV double labeled neurons were also detected in the s. oriens of CA1 (52%), CA2 (27%) and CA3 (36%) subfields. The expression of GFR alpha-1 in principal neurons and in a specific sub-population of GABAergic neurons (PV-containing neurons) suggest that GDNF might modulate, in a selective manner, functions of the entire adult hippocampus.

Animals↗
PubMed2002

The co-expression of ASIC3 with calcitonin gene-related peptide and parvalbumin in the rat trigeminal ganglion.

The co-expression of ASIC3 with calcitonin gene-related peptide (CGRP) or parvalbumin (PV) was examined in the trigeminal ganglion (TG) by a double immunofluorescence method. ASIC3-immunoreactivity (IR) was detected in 23% of TG neurons. These neurons were of various sizes (range= 43-1768 microm(2), mean+/-S.D.=651+/-356 microm(2)); 26% and 14% of ASIC3-immunoreactive (IR) neurons co-expressed CGRP- and PV-immunoreactivity (IR), respectively; 33% and 13% of the TG neurons retrogradely labeled from the tooth pulp and facial skin, respectively, exhibited ASIC3-IR; 36% of CGRP-IR TG neurons which innervate these tissues co-expressed ASIC3-IR. Only 4% of ASIC3-IR cutaneous TG neurons showed PV-IR, while 25% of ASIC3-IR tooth pulp neurons were also immunoreactive for PV. The present study suggests that ASIC3-IR TG neurons supply the tooth pulp and facial skin with unmyelinated or finely myelinated axons. ASIC3-IR neurons which have large myelinated axons may be common in the tooth pulp but not the facial skin. The axonal morphology of ASIC3-IR TG neurons may depend on the variety of their receptive fields.

Acid Sensing Ion Channels↗
PubMed2003

Discrimination of striatopallidum and extended amygdala in the rat: a role for parvalbumin immunoreactive neurons?

Synaptic effects of parvalbumin-immunoreactive (-ir) interneurons (PVs) upon medium spiny neurons may be essential to neural processing in the striatum and, in effect, may serve as an additional feature distinguishing striatum from extended amygdala. The present immunohistochemical study in the rat was done to evaluate the distributions of PVs in the striatum and extended amygdala. Numerous PVs occupy all structures currently regarded as having a striatal composition, including the caudate-putamen, nucleus accumbens, and olfactory tubercle, as well as structures that receive outputs from these, including the globus pallidus, ventral pallidum, entopeduncular nucleus and substantia nigra reticulata. The morphologies of striatal PVs and their distribution were similar to what has been previously reported. In addition, we found that the density of larger neostriatal PVs with extensive and densely immunoreactive dendritic and local axonal arbors is greatest laterally, particularly in striatal districts with slight calbindin-ir, including the striatal patch compartment. In contrast to the situation in striatum, few PVs were observed in the central and medial divisions of the extended amygdala, including the bed nucleus of stria terminalis, interstitial nucleus of the posterior limb of the anterior commissure and central and medial nuclei of the amygdala, or in mesopontine, peribrachial and medullary structures that receive extended amygdala output. The paucity of PVs may be a characteristic feature distinguishing extended amygdala and its projection areas from striatopallidum, as well as the general character of neural processing that occurs in each.

Amygdala↗
PubMed1997

Parvalbumin disappears from GABAergic CA1 neurons of the gerbil hippocampus with seizure onset while its presence persists in the perforant path.

Mongolian gerbils are epilepsy prone animals, a trait observable at the behavioural level during the 2nd month of life. As a unique species difference, gerbils express the calcium-binding protein parvalbumin (PV) in the perforant path from the entorhinal cortex to the hippocampus. In this study, we determined the time of appearance of PV in the layer II neurons of the entorhinal cortex and the perforant path terminals in gerbils between post-natal days 30 and 50. Signs of low grade seizures were observed in few animals from P40 onward. PV stain in the entorhinal cortex and perforant path terminals was already detectable at P30, well before the onset of behavioural seizures and did not change with age. It is suggested that the presence of PV in this pathway may be related to the generation early in life of an epileptogenic focus in the limbic forebrain. Altered inhibitory hippocampal circuits have also been suggested as a cause of seizures in the gerbil. Therefore, we quantitated hippocampal GABA-immunoreactive neurons and the PV-immunoreactive subpopulation. A group of gerbils with a high density of stained pyramidal interneurons in CA1 and one lacking PV-stained perikarya could be distinguished at P40 and P50. The density of GABA-immunoreactive nerve cells however, remained the same in both groups and through the ages studied. Thus, perikaryal PV is lost from intact GABAergic nerve cells at the same time as behavioural seizures are observed. The loss of PV from GABAergic neurons may affect their functional properties and be instrumental for the maintainance of behavioural seizures.

Age of Onset↗
PubMed1999

Long-lasting c-fos and NGF mRNA expressions and loss of perikaryal parvalbumin immunoreactivity in the development of epileptogenesis after ethacrynic acid-induced seizure.

A single cerebroventricular injection of ethacrynic acid (EA), a Cl(-)-ATPase inhibitor, induces generalized tonic-clonic convulsions in mice. To clarify whether such convulsive stimulus triggers a long-lasting rearrangement of the neural circuitry culminating in seizure susceptibility, we examined molecular, cellular and behavioral changes following the EA-induced seizure. The expression of immediate early gene c-fos mRNA as an index for cellular activation increased biphasically, with an early transient increase at 60 min and a late prolonged increase on the 10th to 14th day post-EA administration, most remarkably in the hippocampus and pyriform cortex. On the 14th day post-EA seizure, subconvulsive dose of kainic acid (5-17.5 mg/kg) caused severe (stage 5) seizure in 77% of the mice, with 70% mortality. In addition, the expression of nerve growth factor (NGF) also showed biphasic increases with close spatiotemporal correlation with c-fos expression. Moreover, the number of cell somata and the density of axon fibers of parvalbumin (PARV)-positive cells, a subpopulation of GABAergic interneurons, decreased in area dentata, CA1 and CA3 on the 7th and 14th day post-EA seizure. In area dentata and CA1, the density of glutamic acid decarboxylase (GAD)-positive cells also decreased on the 14th day. Thus, the transient EA-induced seizures appear to develop seizure susceptibility by causing damage of a subpopulation of inhibitory interneurons along with increases in the expression of c-fos and NGF in limbic structures.

Animals↗
PubMed1999

Phosphacan immunoreactivity is associated with perineuronal nets around parvalbumin-expressing neurones.

A special feature of the extracellular matrix in adult brains of various species is the concentration of certain components around different sub-populations of neurones, giving rise to net-like structures termed perineuronal nets. Recently, some of these components have been identified but the function of these nets has yet to be resolved. Using immunofluorescence microscopy, we report here that phosphacan, a chondroitin sulphate proteoglycan, is an additional component of Wisteria floribunda labelled perineuronal nets surrounding parvalbumin-expressing neurones in rat cerebral cortex. Glycoproteins such as tenascin-C and -R have been identified in perineuronal nets and the present detection of phosphacan immunoreactivity in the same entity is of potential physiological importance because of their previously described interactions.

Animals↗
PubMed1999

Calbindin 28kD and parvalbumin immunoreactive neurons receive different patterns of synaptic input in the cat superior colliculus.

Recent evidence suggests that neurons containing the calcium binding proteins calbindin 28kD (CB) and parvalbumin (PV) have differing distributions which match respectively the distribution of W and Y retinal ganglion cell inputs to the cat superior colliculus (SC). In this study we have used electron microscope immunocytochemistry to study directly the synaptic inputs to neurons containing CB and PV. Aspiration lesions of areas 17-18 of visual cortex were made 4 days prior to sacrifice in order to identify degenerating cortical terminals (CT). Retinal terminals (RTs) were identified by their characteristic morphology including large round synaptic vesicles and pale mitochondria. We photographed RTs and CTs that were in contact with immunoreactive profiles sampled in both the superficial gray and optic layers (ol) of SC. CB immunoreactive (ir) dendrites were usually of small to medium caliber and were found to receive synaptic input from RTs. These RTs were all small profiles forming a single synaptic contact with asymmetric densifications. CBir profiles also received other synaptic input, including from terminals with dark mitochondria that contained flattened synaptic vesicles (F profiles). No CBir dendrites were found to receive CT input even though degenerating CTs were found in the vicinity of CBir profiles. By contrast, both RT and CT were found to contact PVir dendrites. RT terminals contacting PVir dendrites were both small and larger profiles with round synaptic vesicles and asymmetric synaptic densifications. CT were undergoing electron dense degeneration but still sometimes formed asymmetric synaptic densifications with PV neurons. PV cells also received F profile synaptic input. We conclude that CB neurons receive small RT synapses that are probably of W origin, while PV neurons receive both RT and CT synapses which are likely related to the Y pathway.

Animals↗
PubMed2003

Age-related changes in calbindin-D28k, calretinin, and parvalbumin-immunoreactive neurons in the human cerebral cortex.

Calbindin-D(28k) (CB), calretinin (CRT), and parvalbumin (PV) are high-affinity cytosolic calcium (Ca(2+)) binding proteins (CBP) that have been found to regulate intracellular calcium concentrations in neurons through their buffering capacity and to protect neurons from insults that induce elevations of intracellular Ca(2+). In earlier studies we observed a substantial and neurochemically specific loss of CB from the human basal forebrain cholinergic neurons (BFCN) in the course of normal aging. In the present experiments we expanded our investigation of age-related changes in calcium binding proteins in the human brain by investigating the status of CB-, CRT-, and PV-positive neurons in 17 cortical areas. There was a trend toward a decrease in the number of CB-immunoreactive neurons in all areas studied. However, this trend reached significance in only 4 areas in which the loss of CB-positive neurons ranged between 20 and 46%. Immunoreactivity for CRT was also decreased in many areas and this difference reached significance in three regions (26-37%). Cortical neurons displaying PV immunoreactivity did not show an age-related change. Comparison with other neurochemically specific cortical neurons indicated a similar age-related loss of nonphosphorylated neurofilament and NADPH-d activity in only a few cortical areas. In contrast, neuronal acetylcholinesterase activity was increased in a few cortical areas. These observations indicate that loss of CBP-positive neurons occurs in restricted cortical regions and is not a specific change as other neurochemically specific neurons also display restricted age-related changes. Furthermore, the age-related changes in cortical CBP-positive neurons appear to be considerably smaller than similar changes in the BFCN. The age-related depletion of CBPs is likely to deprive neurons from the capacity to buffer intracellular calcium and thus to leave them vulnerable to pathological processes that can cause increased intracellular calcium and lead to their degeneration.

Acetylcholinesterase↗
PubMed1983

X-ray absorption near edge structure (XANES) determination of calcium sites of troponin C and parvalbumin.

Using synchrotron radiation at the Frascati storage ring ADONE, the X-ray Absorption Near Edge Structure (XANES) has been applied to determine homologies and modifications of the local structure of the calcium binding sites of troponin C. In all four calcium binding sites, Ca2+ appears to be co-ordinated to carboxyl and carbonyl groups in a characteristic configuration. No structural difference has been found between high and low-affinity sites. A distortion of the Ca2+ site geometry by binding of Mg2+ has been observed. The XANES of parvalbumin has been measured and found to be different from troponin C. A tentative identification of the characteristic XANES spectra of the two different Ca2+ sites in this protein is reported.

Amino Acid Sequence↗
PubMed1985

The functional nature of calcium binding units in calmodulin, troponin C and parvalbumin.

Examination of the interaction of major tranquilizers with calmodulin results in the generalization that the functional nature of calcium binding helix-loop-helix regions found in several calcium binding proteins including calmodulin, troponin C and parvalbumin is dependent upon the topography of the hydrophobic and hydrophilic regions on the amphiphilic N-terminal alpha-helix of the helix-loop-helix conformation formed by the binding of the calcium cation to these proteins. The relation of the topography of this amphiphilic alpha-helix to drug binding is delineated at the molecular level and the results obtained are used to describe the interaction of beta-endorphin, dynorphin, alpha-MSH and other peptides with calmodulin. The utility of this hypothesis is further demonstrated by the description of a possible interaction between troponin C, troponin I and troponin T of the troponin complex in skeletal muscle.

Amino Acid Sequence↗
PubMed1990

Immunocytochemical demonstration of the calcium-binding proteins calbindin-D 28k and parvalbumin in the subiculum, hippocampus and dentate area of the domestic pig.

The distribution of the calcium-binding proteins calbindin-D 28k (CaBP) and parvalbumin (PV) in the hippocampal region of the domestic pig was demonstrated by immunocytochemistry. Scattered CaBP-immunoreactive cell bodies were present in the subiculum, stratum oriens, pyramidal cell layer and stratum radiatum of the hippocampal regio superior and inferior, and the outer plexiform layer and outer hilar cell layer of the dentate hilus. Other cell bodies and bundles of stained fibers were present in stratum moleculare of regio superior and inferior, and in the outer third of the molecular layer of the fascia dentata. Terminal-like CaBP-immunoreactivity was seen in the subiculum and around cell bodies in the pyramidal cell layer of regio superior and inferior and the dentate granular cell layer. Scattered PV-immunoreactive cell bodies were present in stratum oriens and the pyramidal cell layer of regio superior and inferior, and in the outer plexiform layer and outer hilar cell layer of the dentate hilus. Terminal-like PV-immunoreactivity surrounded the cell bodies in the pyramidal cell layer of regio superior and inferior and in the dentate granular cell layer. The distribution of CaBP and PV in the pig hippocampus is compared to that of other more commonly used experimental animals. Whereas the distribution of PV-immunoreactivity in the pig hippocampus appears identical to that of the rat hippocampus, the distribution of CaBP-immunoreactivity in the pig hippocampus differs markedly from that of the rat hippocampus, the most prominent feature being a lack of CaBP-immunoreactivity in the granule cells, mossy fibers and pyramidal cells in the pig. The functional implications of calcium-binding proteins in the brain are discussed.

Animals↗
PubMed2002

The thalamic reticular and perireticular nuclei in developing rabbits: patterns of parvalbumin expression.

Due to its strategic position, the thalamic reticular nucleus (TRN) plays an important role within the thalamo-cortical circuits. The perireticular thalamic nucleus (PRN) is a smaller group of cells, which is associated with the TRN and lies among the fibres of the internal capsule (IC). Studies of nuclei in rodents and carnivores have been conducted employing a number of different tools. The use of calcium-binding proteins is one example. It needs to be noted that rabbits have been regarded as intermediate between rodents and carnivores in relation to local GABAergic circuits. In the present study, sections from rabbits at different ages (prenatal, postnatal and adult) were examined to determine the parvalbumin (PV) expression in the developing TRN and PRN. In the TRN, there is one wave of PV expression during development, from caudal parts of the nucleus towards the rostral pole. At E22 there is already an incipient PV expression. In the adult stage, the TRN is completely positive to PV. The present study clearly indicates the presence of the PRN in the developing rabbit. The first PV positive cells were visible at E24, meanwhile the immunoreactivity was at its maximum at early postnatal stages (P0-P8). Two different types of perireticular cells in the IC were identified and the changes concerning neuronal morphology and orientation were described. The comparison between these results and previous data obtained in rats, ferrets or cats suggest that rabbits could represent an intermediate stage in the evolution of thalamic circuits and could be considered as useful neurobiological model.

Animals↗
PubMed1997

Retrosplenial/presubicular continuum in primates: a developmental approach in fetal macaques using neurotensin and parvalbumin as markers.

In spite of numerous hodological and neuropsychological studies emphasizing the multimodal connections and integrative functions of the retrosplenial cortex in primates, the precise fate of its caudoventral extent and the composition of the merging area with the hippocampal formation remain a matter of debate. We reported previously how the anlage of the retrosplenial cortex merges with the immature presubicular zone in the fetal rhesus monkey at the end of the first trimester of gestation. In the present study, this caudal area was further defined on a chemoarchitectonic basis, particularly during the late prenatal and perinatal stages, which correspond to the development of the cingulate sulcus and temporal gyri, and the differentiation of the retrosplenial/subicular complex. Neurotensin (NT), a pyramidal cell marker in the limbic cortex, and parvalbumin (PV), a marker of a subset of inhibitory local circuit neurons in the hippocampal formation, were used as immunocytochemical markers. According to distinct chemoarchitectural patterns, (1) areas 29 l and 29 m of the retrosplenial cortex formed a triangle-shaped ventral expansion which merged with a similar but dorsal expansion of the pre/parasubicular fields. A temporal extension of area 29 m down to area TH could not be detected. The pre/parasubiculum contributed with area 29 m to the lateral bank of the calcarine sulcus as far as the most caudal extent of the hippocampal formation. (2) The lamina principalis interna of the presubiculum was well individualized and did not appear as a simple horizontal shift of adjoining fields. (3) NT and PV displayed a distinct temporal profile of development. NT was already expressed in the pyramidal cells of the prospective retrosplenial cortex and ventral hippocampal formation at E47 (term 165 days). Major pathways of the hippocampal formation and retrosplenial cortex (fimbria, fornix, angular and cingulum bundles) were progressively labeled indicating early developing projections. A large set of NT-positive afferents reached the retrosplenial cortex between E114 and E120. Their laminar distribution was compatible with a thalamic or a subicular origin. (4) The development of PV expression was delayed until the last quarter of gestation, supporting its proposal as a signal of functional onset. The developmental fate and the particular connections of the presubiculum suggest that its functional importance should be further investigated during infancy and adulthood.

Animals↗