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PubMed1988

Two distinct phases and mechanisms of axonal growth shown by primary vestibular fibres in the brain, demonstrated by parvalbumin immunohistochemistry.

Antibodies to parvalbumin label intensely a small number of non-overlapping fibre systems in embryonic rat brain. All are in hindbrain--the oculomotor and trochlear motor fibres, the acoustic and vestibular fibres of the VIIIth nerve, and an unidentified group of fibres which ascend under the dorsal surface in caudal medulla. Of these, the vestibular fibres are the first to acquire parvalbumin immunoreactivity, and we have used this property to follow the growth of their axons in the brain. This occurs in two phases. In the first, occurring at embryonic days 12-14, the axons grow in small groups or fascicles under the pial surface to their most distant terminal zones rostrally in the cerebellum and caudally in the descending vestibular nuclei. This growth is directed towards the two sites where germinal neuroepithelium is expanding over the medullary velum in forming the cerebellum and lateral recess of the IVth ventricle. In a second stage, commencing at E15, individual collaterals branch from these fascicles to arborize amongst their presumptive synaptic targets (cells of the vestibular nuclei and vestibulocerebellum) located in the sub-ventricular and ventricular layers. In this phase the axons follow a radial route, at right angles to their original subpial course, possibly by growing along radial glial processes. The target cells then migrate to their final position with the vestibular axons maintaining contact with them. The vestibular fibres are the first axons to enter the cerebellum, but from E15 onwards their fascicles are joined by increasing numbers of non-vestibular fibres following the same course. These other axons, and the movement of cells to form the deep cerebellar nuclei, separate the fascicles of vestibular fibres so that their course into the cerebellum becomes very diffuse. Thus this single set of axons grow, not only in two distinct phases, but also follow distinctly different substrates for growth in each. Furthermore, they then appear to act as pioneer fibres guiding the entry or egress of later-developing axons to or from the cerebellum.

Animals↗
PubMed1991

Parvalbumin-positive neurons in rat dorsal hippocampus contain muscarinic acetylcholine receptors.

The present study describes the colocalization of muscarinic acetylcholine receptors (mAChRs) and the calcium-binding protein parvalbumin (PARV) in nonpyramidal neurons of the rat dorsal hippocampus by means of dual-label immunocytochemistry. Fifty-two percent of all muscarinic cholinoceptive nonpyramidal neurons contain parvalbumin. Conversely, the vast majority (88.4%) of all PARVergic neurons possess mAChRs. The PARVergic neurons embedded within the CA1 pyramidal cell layer are nearly all (98.9%) immunopositive for mAChRs. These results indicate that the cholinergic septohippocampal projection represents a major afferent system upon the inhibitory PARVergic neurons.

Animals↗
PubMed1994

Distribution of calretinin, calbindin-D28k, and parvalbumin in the rat thalamus.

The localization of three calcium-binding proteins, calretinin, calbindin-D28k, and parvalbumin, in the rat thalamus was immunohistochemically examined. a) Some thalamic regions revealed cells almost exclusively containing one of the calcium-binding proteins. For example, almost only calretinin-stained cells were found in the central medial and paraventricular nuclei. Calbindin-D28k-stained cells were mostly found in the centrolateral, interanteromedial, anteromedial, and posterior nuclei. Only parvalbumin-positive cells were found in the central part of the reticular nucleus. b) Other regions expressed overlap between the distributions of two cell components composed of different calcium-binding proteins. For example, both calretinin-stained cells and calbindin-D28k-labeled cells were found in the lateroposterior, intermediodorsal, rhomboid, and reuniens nuclei. c) Other regions showed no cells stained for any of the calcium-binding proteins. For example, generally no calcium-binding protein was detected in neurons of the anterodorsal, anteroventral, ventrolateral, ventral posterolateral, ventral posteromedial, or gelatinosus nuclei, or of the central part of the mediodorsal nucleus. These three proteins serve as useful marker for localizing subpopulations of neurons within the thalamus.

Animals↗
PubMed1994

AF64A affects septal choline acetyltransferase but not parvalbumin immunoreactive cells.

Rats received bilateral intracerebroventricular (ICV) infusions of either AF64A (1.5 nmol/ventricle; n = 9) or vehicle (3.0 microliters/ventricle; n = 7). Four weeks later, the animals were anesthetized and their brains processed to visualize and quantify choline acetyltransferase (ChAT) immunoreactive (IR) and parvalbumin-IR GABAergic neurons in the septal complex by immunocytochemistry (PAP method). AF64A significantly reduced the number of ChAT-IR perikarya in the medial septum (28%), ventral limb of the diagonal band of Broca (30%), and horizontal limb of the diagonal band of Broca (20%), but did not affect the number of parvalbumin-containing GABAergic neurons in any of the septal subdivisions. These results provide further evidence that AF64A is a selective cholinotoxin.

Animals↗
PubMed1994

Partial colocalization of the GABAA receptor with parvalbumin and calbindin D-28K in neurons of the visual cortex and the dorsal lateral geniculate nucleus of the cat.

Monoclonal antibodies to a synthetic peptide fragment of the beta 1-subunit of the bovine central GABAA/benzodiazepine receptor were used to investigate immunocytochemically the distribution of this receptor in the visual system of the cat. Labeled neurons were observed in all layers of the visual cortex and the dorsal lateral geniculate nucleus. About half of the total cortical or geniculate neuronal population was found to be positive. To further identify immunocytochemically these GABAA receptor expressing cells, double stainings were undertaken with, on one hand, the monoclonal antibodies directed against the receptor complex, and on the other hand polyclonal antisera directed against cat muscle parvalbumin or chicken calbindin D-28K. A high degree of colocalization between either of the two calcium binding proteins and the GABAA receptor was found in the upper layers (I, II and III) of the visual cortex and in the A and C laminae of the dorsal lateral geniculate nucleus; all calbindin D-28K-positive cells were immunoreactive for the GABAA receptor. The parvalbumin-positive cells, scattered throughout all layers of the dorsal lateral geniculate nucleus and the visual cortex, except cortical layer I, were also all positive for the GABAA receptor. However, a large proportion of all GABAA receptor bearing cells were negative for one of the calcium binding proteins.

Animals↗
PubMed1996

Distribution of neurofilament protein and calcium-binding proteins parvalbumin, calbindin, and calretinin in the canine hippocampus.

Neurofilament protein and calcium-binding proteins parvalbumin, calbindin, and calretinin are present in morphologically distinct neuronal subpopulations in the mammalian cerebral cortex. Immunohistochemical studies of the hippocampal formation and neocortex have demonstrated that while neurofilament protein and calbindin are localized in subsets of pyramidal neurons, the three calcium-binding proteins are useful markers to differentiate non-overlapping populations of interneurons. To date, most studies have been performed in rodents and primates. In the present analysis, we analyzed the distribution of these proteins in the canine hippocampus. Neurofilament protein was present in large multipolar neurons in the hilus and in pyramidal neurons in the CA3 field, whereas pyramidal neurons in the CA1 field and subiculum were less intensely immunoreactive. Parvalbumin immunoreactivity was observed in large multipolar neurons in the hilus and throughout the CA3-CA1 fields, in a few pyramidal-shaped neurons in the CA1 field and subiculum, and had a distinct neuropil staining pattern in the granule cell layer and stratum pyramidale of the Ammon's horn. Calbindin immunoreactivity displayed a strong labeling of the granule cells and mossy fibers and was also observed in a population of moderately immunoreactive neurons in the CA1 field and subiculum. Calretinin immunoreactivity was relatively weaker overall. The inner molecular layer in the dentate gyrus had a distinct band of labeling, the stratum lacunosum/moleculare contained a punctate neuropil staining, and there were a few small multipolar neurons in the hilus, CA3-CA1 fields, and subiculum. Comparison of the staining patterns observed in the dog hippocampus with those in human, macaque monkeys and rats revealed that although there are some subregional differences among these taxa, the dog may constitute a valuable large animal model for the study of certain neurological conditions that affect humans, in spite of the phylogenetic distance between carnivores and primates.

Animals↗
PubMed2004

Neonatal neurosteroid administration alters parvalbumin expression and neuron number in medial dorsal thalamus of adult rats.

The neuroactive steroid 3alpha-hydroxy-5alpha-pregnane-20-one (allopregnanolone) is a potent endogenous modulator of GABAA receptor function. A single neonatal allopregnanolone administration (10 mg/kg, i.p.) was previously shown to alter the localization of parvalbumin-positive neurons in the prefrontal cortex at maturity. Cells in the prefrontal cortex receive the majority of their inputs from the medial dorsal nucleus of the thalamus. We investigated whether neonatal allopregnanolone administration alters the neuronal population of the medial dorsal nucleus of the thalamus. We show that the number of parvalbumin-expressing neurons is increased while the total neuron number is decreased in the medial dorsal nucleus after allopregnanolone administration. EAAT3 (excitatory amino acid transporter type 3, the neuron-specific glutamate reuptake transporter) immunoreactivity was unchanged in adjacent sections. These findings suggest that neonatal allopregnanolone administration disrupts the normal development of the prefrontal cortex and medial dorsal thalamus, indicating that neurosteroid levels are important for proper development of thalamocortical systems and may play a role in neurodevelopmental disorders such as schizophrenia.

Amino Acid Transport System X-AG↗
PubMed2006

Prenatal acoustic stimulation influences neuronal size and the expression of calcium-binding proteins (calbindin D-28K and parvalbumin) in chick hippocampus.

Prenatal auditory enrichment by species-specific sounds and sitar music enhances the expression of immediate early genes, synaptic proteins and calcium binding proteins (CaBPs) as well as modifies the structural components of the brainstem auditory nuclei and auditory imprinting area in chicks. There is also facilitation of postnatal auditory preference of the chicks to maternal calls following both types of sound stimulation indicating prenatal perceptual learning. To examine whether the sound enrichment protocol also affects the areas related to learning and memory, we assessed morphological changes in the hippocampus at post-hatch day 1 of control and prenatally sound-stimulated chicks. Additionally, the proportions of neurons containing calbindin D-28K and parvalbumin immunoreactivity as well as their protein levels were determined. Fertilized eggs of domestic chick were incubated under normal conditions of temperature, humidity, forced draft of air as well as light and dark (12:12h) photoperiods. They were exposed to patterned sounds of species-specific and sitar music at 65 dB for 15 min per hour over a day/night cycle from day 10 of incubation till hatching. The hippocampal volume, neuronal nuclear size and total number of neurons showed a significant increase in the music-stimulated group as compared to the species-specific sound-stimulated and control groups. However, in both the auditory-stimulated groups the protein levels of calbindin and parvalbumin as well as the percentage of the immunopositive neurons were increased. The enhanced proportion of CaBPs in the sound-enriched groups suggests greater Ca(2+) influx, which may influence long-term potentiation and short-term memory.

Acoustic Stimulation↗
PubMed2006

Potential influence of Asp in the Ca2+ coordination position 5 of parvalbumin on the calcium-binding affinity: a computational study.

Parvalbumins (PV) are calcium-binding proteins, all sharing the common helix-loop-helix (EF-hand) motif. This motif contains a central twelve-residue Ca(2+)-binding loop with the flanking helices positioned roughly perpendicular to each other. The precise role of these coordination residues has been the subject of intense studies. In this work, we focus on the coordination position 5 in the CD Ca(2+)-binding site of silver hake parvalbumin isoform B (SHPV-B). The most common residue at site 5 of calcium-binding loop in canonical EF-hands is Asp [B.J. Marsden, G.S. Shaw, B.D. Sykes, Biochem. Cell Biol. 68 (1990) 587-601], but in the CD site of PV, this position is almost always serine (Ser). The substitution of Ser with Asp will add the 5th carboxylate residue in the CD coordination sphere. However, as predicted by the acid pair hypothesis, the Ca(2+)-binding affinity would be maximized in an EF-hand motif that has four carboxylate ligands paired along the +/-x, and +/-z-axes [R.E. Reid, R.S. Hodges, J. Theor. Biol. 84 (1980) 401-444]. Molecular dynamics simulations and free energy calculations were employed to investigate the influence of Ser to Asp mutation at position 5 on calcium-binding affinity. We found that the Asp variant exhibited remarkable stability during the entire molecular dynamics simulation, with not only the retention of the Ca(2+)-binding site, but also increased compactness in the coordination sphere. The S55D fragment also accommodated Ca(2+) well. We conclude that the reason why Asp which is the most common residue at site 5 of calcium-binding loop in canonical EF-hands has never been identified at this position experimentally for PVs might be related to its physiological functions.

Amino Acid Sequence↗
PubMed2004

Perinatal flunitrazepam exposure causes persistent alteration of parvalbumin-immunoreactive interneuron localization in rat prefrontal cortex.

GABA regulates proliferation via GABAA receptors during development of the neocortex. We recently demonstrated that the endogenous GABAA receptor modulator allopregnanolone plays a role in regulating normal neurodevelopment in prefrontal cortex. Benzodiazepine exposure during early development produces marked behavioral changes in adult rats. To determine if exposure to benzodiazepines during development alters GABAergic interneurons in prefrontal cortex (PFC), rat pups were exposed to flunitrazepam (2.5 mg/kg) on postnatal day (P) 2 and assayed for parvalbumin- and calbindin-immunoreactivity on P80. The ratio of parvalbumin labeled cells in deep vs. superficial layers increased five-fold; calbindin-immunoreactivity and total cell number were not altered. These data are consistent with altered distribution of a subset of interneurons after benzodiazepine exposure and suggest a role for GABAA receptor modulation in normal development of GABAergic systems in PFC.

Animals↗
PubMed2003

Estimation of parvalbumin Ca(2+)- and Mg(2+)-binding constants by global least-squares analysis of isothermal titration calorimetry data.

The use of competitive isothermal titration calorimetry (ITC) to measure high-affinity binding constants has been largely restricted to systems with a single binding site or multiple identical sites. This study demonstrates the extension of this approach to proteins with two nonequivalent EF-hand Ca(2+)-binding sites--rat beta parvalbumin and the S55D/E59D variant of rat alpha parvalbumin. The method involves simultaneous (global) least-squares analysis of titrations with Ca(2+), with Mg(2+), with Ca(2+) in the presence of Mg(2+), and with Ca(2+) or Mg(2+) in the presence of a competitive chelator (EDTA or EGTA). The Ca(2+) and Mg(2+) binding constants obtained for rat beta agree well with estimates obtained by flow dialysis. Although the Ca(2+) affinity of alpha S55D/E59D is too high to measure by flow dialysis, it was amenable to analysis using the ITC-based approach. The combined S55D and E59D mutations increase the Ca(2+) and Mg(2+) affinities of the mutated binding site by factors of 14 and 26, respectively. This behavior is consistent with that seen previously for the rat beta S55D variant.

Amino Acid Substitution↗
PubMed1989

Avian thymic hormone (ATH) is a parvalbumin.

Amino acid sequence analysis of a protein from chicken thymus tissue which promotes immunological maturity in chicken bone marrow cells in culture has established sequences of a 45-residue fragment, a 24-residue fragment and a 9-residue and an 8-residue peptide. Independent comparison of the 45- and 24-residue fragments with known amino acid sequences by computer search has unequivocally identified avian thymic hormone as a parvalbumin. This is the first demonstration that a protein previously identified by a biological function is a parvalbumin.

Amino Acid Sequence↗
PubMed2002

Molecular mechanisms of calcium and magnesium binding to parvalbumin.

Molecular dynamics simulations have been used to investigate the relationship between the coordinating residues of the EF-hand calcium binding loop of parvalbumin and the overall plasticity and flexibility of the protein. The first simulation modeled the transition from Ca(2+) to Mg(2+) coordination by varying the van der Waals parameters for the bound metal ions. The glutamate at position 12 could be accurately and reversibly seen to be a source of selective bidentate ligation of Ca(2+) in the simulations. A second simulation correlated well with the experimental observation that an E101D substitution at EF loop position 12 results in a dramatically less tightly bound monodentate Ca(2+) coordination by aspartate. A final set of simulations investigated Ca(2+) binding in the E101D mutant loop in the presence of applied external forces designed to impose bidentate coordination. The results of these simulations illustrate that the aspartate is capable of attaining a suitable orientation for bidentate coordination, thus implying that it is the inherent rigidity of the loop that prevents bidentate coordination in the parvalbumin E101D mutant.

Amino Acid Sequence↗
PubMed1981

The time-course of Ca2+ exchange with calmodulin, troponin, parvalbumin, and myosin in response to transient increases in Ca2+.

We have modeled the time-course of Ca2+ binding to calmodulin, troponin, parvalbumin, and myosin in response to trains of transient increases in the free myoplasmic calcium ion concentration (pCa). A simple mathematical expression was used to describe each pCa transient, the shape and duration of which is qualitatively similar to those thought to occur in vivo. These calculations assumed that all individual metal binding sites are noninteracting and that Ca2+ bind competitively to the Ca2+-Mg2+ sites of troponin, parvalbumin, and myosin. All the on-and-off rate constants for both Ca2+ and Mg2+ were obtained either from the literature or from our own research. The percent saturation of the Ca2+-Mg2+ sites with Ca2+ was found to change very little in response to each pCa transient in the presence of 2.5 X 10(-3)M Mg2+. Our analysis suggests that the Ca2+ content of these sites is a measure of the intensity and frequency of recent muscle activity because large changes in the Ca2+ occupancy of these sites can occur with repeated stimulation. In contrast, large rapid changes in the amount of Ca2+ bound to the Ca2+-specific sites of troponin and calmodulin are induced by each pCa transient. Thus, only sites of the "Ca2+-specific" type can act as rapid Ca2+-regulatory sites in muscle. Fluctuation in the total amount of Ca2+ bound to these sites in response to various types of pCa transients further suggests that in vivo only about one-half to one-third of the total steady-state myofibrillar Ca2+-binding capacity exchanges Ca2+ during any single transient.

Animals↗
PubMed2001

Parvalbumin-positive projection neurons characterise the vocal premotor pathway in male, but not female, zebra finches.

Parvalbumin (PV) and calbindin (CB) immunoreactivities were assessed in nucleus robustus archistriatalis (RA) of male and female zebra finches, together with retrograde labelling of RA neurons. The results of double and triple labelling experiments suggested that, in males, moderately and faintly PV-positive neurons were projection neurons, but that all intensely PV-positive cells were not. The latter, which are presumably interneurons, were also intensely CB-positive, and may correspond to the GABAergic inhibitory interneurons identified by others. In addition, the complete RA pathway and its terminal fields in the respiratory-vocal nuclei of the brainstem were strongly PV-positive. In female zebra finches, which do not sing, no evidence was found that PV-positive RA cells were projection neurons, yet the pattern of projections of RA neurons, as determined by anterograde transport of biotinylated dextran amine, was very similar to that of RA in males. Moreover, in females, RA neurons retrogradely labelled from injections of cholera toxin B-chain into the tracheosyringeal nucleus (XIIts) were abundant and included, in the lateral part of the nucleus, a population of cells that were as large as those in the male RA. Parvalbumin immunoreactivity was also present in RA and its projections in males of several other songbird species (northern cardinal, brown headed cowbird, canary) and in the female cardinal, which sings to some extent, but the labelling was not as intense as that in male zebra finches.

Animals↗
PubMed2002

Parvalbumin neuron circuits and microglia in three dopamine-poor cortical regions remain sensitive to amphetamine exposure in the absence of hyperthermia, seizure and stroke.

The dopamine-releasing and depleting substance amphetamine (AMPH) can make cortical neurons susceptible to damage, and the prevention of hyperthermia, seizures and stroke is thought to block these effects. Here we report a 2-day AMPH treatment paradigm which affected only interneurons in three cortical regions with average or below-average dopamine input. AMPH (six escalating doses/day ranging from 5 to 30 mg/kg for 2 days) was given at 17-18 degrees C ambient temperature (T) to adult male rats. During the 2-day AMPH treatment, peak body T stayed below 38.9 degrees C in 40% of the AMPH treated rats. In 60% of the rats, deliberate cooling suppressed (<39.5 degrees C) or minimized (<40.0 degrees C) hyperthermia. Escalation of stereotypes to seizure-like behaviors was rare and post-mortem morphological signs of stroke were absent. Neurons labeled with the anionic, neurodegeneration-marker dye Fluoro-Jade (F-J) were seen 1 day after dosing, peaked 3 days later, but were barely detectable 14 days after dosing. Only nonpyramidal neurons in layer IV of the somatosensory barrel cortex and in layer II of the piriform cortex and posterolateral cortical amygdaloid nucleus were labeled with Fluoro-Jade. Isolectin B-labeled activated microglia were only detected in their neighborhood. F-J labeled neurons were extremely rare in cortical regions rich in dopamine (e.g. cingulate cortex), and were absent in cortical regions with no dopamine (e.g. visual cortex). Parvalbumin was seen in some Fluoro-Jade-labeled neurons and parvalbumin immunostaining in local axon plexuses intensified. This AMPH paradigm affected fewer cortical regions, and caused smaller reduction in striatal tyrosine hydroxylase (TH) immunoreactivity than previous 1-day AMPH regimens generating seizures or severe (above 40 degrees C) hyperthermia. Correlation between peak or mean body T and the extent of neurodegeneration or microgliosis was below statistical significance. Astrogliosis (elevated levels of the astroglia-marker, glial fibrillary acidic protein (GFAP)) was detected in many brain regions. In the striatum and midbrain, F-J labeled neurons and activated microglia were absent, but astrogliosis, decreased TH immunolabel, and swollen TH fibers were detected. In sum, after this AMPH treatment, cortical pyramidal neurons were spared, but astrogliosis was brain-wide and some interneurons and microglia in three cortical regions with average or below-average dopamine input remained sensitive to AMPH exposure.

Amphetamine↗
PubMed1994

Cortical areas are revealed by distribution patterns of proteoglycan components and parvalbumin in the Mongolian gerbil and rat.

Cortical areas in rodents have been basically characterized by its cytoarchitecture, connectivity or by physiological parameters. In this study we show that they are revealed by distribution patterns of proteoglycans and parvalbumin-immunoreactivity. Brains of young adult Mongolian gerbils (Meriones unguiculatus) and Wistar rats were cut into series of transversal sections. Proteoglycan components were detected using the N-acetylgalactosamine binding Wisteria floribunda agglutinin (WFA) and antibodies against chondroitin sulphate proteoglycan (CSPG). Differences between cortical areas were found to exist with regard to the occurrence and the density of perineuronal nets, but were also expressed in varying staining intensities for WFA and CSPG of the neuropil. Primary neocortical areas (somatosensory, auditory, visual cortex) were characterized by an intense neuropil staining in layer IV and the upper part of layer VI. Using the same methods strong labelling was also typical of the neuropil in the retrosplenial cortex, of layer Ia in the prepiriform cortex and the hippocampal CA3 field. In tangential sections cut from gerbil cortical hemispheres, some of the heavily lectin-stained cortical areas were sharply delineated from adjacent faintly labelled regions, others showed more diffuse borders. In the rat, the area-specific staining for WFA was less clearly expressed than in the gerbil. Immunocytochemistry of the calcium-binding protein parvalbumin in alternate sections showed labelling patterns of neuropil which resembled those of WFA-binding and CSPG-immunoreactivity in the entire neocortex and hippocampus. From these results it can be concluded that functional peculiarities of cortical fields may not only be determined by neuronal network parameters but also by the spatial arrangement of extracellular matrix proteoglycans.

Animals↗
PubMed1997

Parvalbumin-containing neurons in the basal forebrain receive direct input from the substantia nigra-ventral tegmental area.

By means of anterograde tracing of Phaseolus vulgaris-leucoagglutinin (PHA-L) it was determined if parvalbumin-immunoreactive neurons in the basal forebrain receive a direct synaptic input from the A9-A10 dopaminergic nuclei of the substantia nigra and ventral tegmental area. Forebrain sections were processed for immunocytochemical detection of PHA-L and parvalbumin (PV) at light and electron microscopic levels. At the ultrastructural level, PHA-L-labeled terminals were found to establish synaptic contacts with PV-immunoreactive neuronal somata in the ventromedial globus pallidus, the ventral pallidum, the internal capsule, and the substantia innominata. PV-containing neurons in pallidal and adjacent basal forebrain territories are thus directly targeted by presumably A9-A10 dopaminergic neurons and represent a novel aspect of midbrain dopaminergic control of basal forebrain neuronal activity.

Animals↗