Role Of Parvalbumin Research Articles

Parvalbumin neurons in the nucleus accumbens shell modulate seizure in temporal lobe epilepsy

A growing number of clinical and animal studies suggest that the nucleus accumbens (NAc), especially the shell, is involved in the pathogenesis of temporal lobe epilepsy (TLE). However, the role of parvalbumin (PV) GABAergic neurons in the NAc shell involved in TLE is still unclear. In this study, we induced a spontaneous TLE model by intrahippocampal administration of kainic acid (KA), which generally induce acute seizures in first 2 h (acute phase) and then lead to spontaneous recurrent seizures after two months (chronic phase). We found that chemogenetic activation of NAc shell PV neurons could alleviate TLE seizures by reducing the number and period of focal seizures (FSs) and secondary generalized seizures (sGSs), while selective inhibition of PV exacerbated seizure activity. Ruby-virus mapping results identified that the hippocampus (ventral and dorsal) is one of the projection targets of NAc shell PV neurons. Chemogenetic activation of the NAc-Hip PV projection fibers can mitigate seizures while inhibition has no effect on seizure ictogenesis. In summary, our findings reveal that PV neurons in the NAc shell could modulate the seizures in TLE via a long-range NAc-Hip circuit. All of these results enriched the investigation between NAc and epilepsy, offering new targets for future epileptogenesis research and precision therapy.

Parvalbumin interneuron activity drives fast inhibition-induced vasoconstriction followed by slow substance P-mediated vasodilation

The role of parvalbumin (PV) interneurons in vascular control is poorly understood. Here, we investigated the hemodynamic responses elicited by optogenetic stimulation of PV interneurons using electrophysiology, functional magnetic resonance imaging (fMRI), wide-field optical imaging (OIS), and pharmacological applications. As a control, forepaw stimulation was used. Stimulation of PV interneurons in the somatosensory cortex evoked a biphasic fMRI response in the photostimulation site and negative fMRI signals in projection regions. Activation of PV neurons engaged two separable neurovascular mechanisms in the stimulation site. First, an early vasoconstrictive response caused by the PV-driven inhibition is sensitive to the brain state affected by anesthesia or wakefulness. Second, a later ultraslow vasodilation lasting a minute is closely dependent on the sum of interneuron multiunit activities, but is not due to increased metabolism, neural or vascular rebound, or increased glial activity. The ultraslow response is mediated by neuropeptide substance P (SP) released from PV neurons under anesthesia, but disappears during wakefulness, suggesting that SP signaling is important for vascular regulation during sleep. Our findings provide a comprehensive perspective about the role of PV neurons in controlling the vascular response.

Role of parvalbumin in fatigue-induced changes in force and cytosolic calcium transients in intact single mouse myofibers.

One of the most important cytosolic Ca2+ buffers present in mouse fast-twitch myofibers, but not in human myofibers, is parvalbumin (PV). Previous work using conventional PV gene (PV) knockout (PV-KO) mice suggests that lifelong PV ablation increases fatigue resistance, possibly due to compensations in mitochondrial volume. In this work, PV ablation was induced only in adult mice (PV-KO), and contractile and cytosolic Ca2+ responses during fatigue were studied in isolated muscle and intact single myofibers. Results were compared with control littermates (PV-Ctr). We hypothesized that the reduced myofiber cytosolic Ca2+ buffering developed only in adult PV-KO mice leads to a larger cytosolic free Ca2+ concentration ([Ca2+]c) during repetitive contractions, increasing myofiber fatigue resistance. Extensor digitorum longus (EDL) muscles from PV-KO mice had higher force in unfused stimulations (∼50%, P < 0.05) and slowed relaxation (∼46% higher relaxation time, P < 0.05) versus PV-Ctr, but muscle fatigue resistance or fatigue-induced changes in relaxation were not different between genotypes (P > 0.05). In intact single myofibers from flexor digitorum brevis (FDB) muscles, basal and tetanic [Ca2+]c during fatiguing contractions were higher in PV-KO (P < 0.05), accompanied by a greater slowing in estimated sarcoplasmic reticulum (SR) Ca2+-pumping versus PV-Ctr myofibers (∼84% reduction, P < 0.05), but myofiber fatigue resistance was not different between genotypes (P > 0.05). Our results demonstrate that although the estimated SR Ca2+ uptake was accelerated in PV-KO, the total energy demand by the major energy consumers in myofibers, the cross-bridges, and SR Ca2+ ATPase were not altered enough to affect the energy supply for contractions, and therefore fatigue resistance remained unaffected.NEW & NOTEWORTHY Parvalbumin (PV) is a cytosolic Ca2+ buffer that is present in mouse myofibers but not in human muscle. We show that inducible knockout of PV leads to increases in myofiber cytosolic free Ca2+ concentrations and slowing of Ca2+ pumping during fatigue versus control mice. However, PV ablation does not interfere with fatigue-induced slowing in relaxation or fatigue resistance. These data support the use of mouse muscle as a suitable model to investigate human muscle fatigue.

Parvalbumin-Containing Enteric Metasympathetic Neurons in Postnatal Ontogeny

The calcium-binding protein parvalbumin is selectively expressed in various subpopulations of neurons in the central and peripheral nervous system. The localization and percentage composition of parvalbumin-immunoreactive neurons were determined in the metasympathetic enteric ganglia of the small and large intestine in rats of different ages (1, 10, 20, 30, and 60 days and two years). Parvalbumin-immunoreactive neurons were present in the intermuscular plexus during ontogeny, from birth to old age, and the percentage of these cells in the intermuscular plexus in the small intestine increased from day 10 to day 20 of life and in that of the large intestine in the first 10 days of life. Parvalbumin was absent from the submucous plexus of the small and large intestine in neonatal rats, but was present from day 10. The percentage of parvalbumin-immunoreactive neurons in the submucous plexus increased between day 10 and day 20 of life. On aging, the proportions of parvalbumin-immunoreactive neurons in the intermuscular and submucous plexuses did not change significantly. Thus, the proportion of parvalbumin-immunopositive neurons in the intramural intestinal ganglia increased in early postnatal ontogeny. This increase is probably associated with the buffer role of parvalbumin in relation to Ca2+ ions.

Parvalbumin Affects Skeletal Muscle Trophism Through Modulation of Mitochondrial Calcium Uptake

Parvalbumin (PV) is a cytosolic EF-hand calcium-binding protein highly expressed in fast skeletal muscle fibers. By acting as a Ca2+ buffer, PV contributes to increase the relaxation rate in skeletal muscle fibers. More recently, PV has been described as one of the “atrogenes”, the collection of genes commonly up- and downregulated during both disuse and systemic types of muscle atrophy. Here we exploited the murine strain lacking PV (PV-/-) to explore the link between PV function as Ca2+ buffer and the control of muscle trophism. First, by using denervation as an acute model of atrophy, we found a lower muscle mass loss in PV-/- than in WT mice. Then, by acutely silencing and overexpressing PV in adult muscles of WT mice, we discovered that its downregulation was accompanied by hypertrophy and its upregulation by atrophy. Using compartment specific Ca2+ probes, we also explored three cellular compartments: cytosol, sarcoplasmic reticulum and mitochondria. The lack of PV was associated with an increased Ca2+ uptake in mitochondria, which could be supported by MCU complex components overexpression and an increased mitochondria volume. Intriguingly, MCU silencing abolished the hypertrophic effect of PV ablation on fiber size, suggesting a mitochondrial Ca2+-dependent mechanism of control of muscle trophism. This is corroborated by the induction of the expression of the mitochondrial Ca2+-dependent PGC1-a4 that promotes skeletal muscle hypertrophy. These data reveal a novel role of PV that links the control of cytosolic Ca2+ concentration to mitochondrial adaptations, ultimately leading to the control of muscle mass.

Parvalbumin expression in oligodendrocyte-like CG4 cells causes a reduction in mitochondrial volume, attenuation in reactive oxygen species production and a decrease in cell processes\u2019 length and branching

Forebrain glial cells - ependymal cells and astrocytes -acquire upon injury- a “reactive” phenotype associated with parvalbumin (PV) upregulation. Since free radicals, e.g. reactive oxygen species (ROS) play a role in the pathogenesis of multiple sclerosis, and that PV-upregulation in glial cells is inversely correlated with the level of oxidative stress, we hypothesized that PV-upregulation might also protect oligodendrocytes by decreasing ROS production. Lentiviral transduction techniques allowed for PV overexpression in CG4 oligodendrocyte progenitor cells (OPCs). Depending on the growth medium CG4 cells can be maintained in an OPC-like state, or induced to differentiate into an oligodendrocyte (OLG)-like phenotype. While increased levels of PV had no effect on cell proliferation and invasiveness in vitro, PV decreased the mitochondria volume in CG4 cell bodies, as well as the mitochondrial density in CG4 processes in both OPC-like and OLG-like states. In line with the PV-induced global decrease in mitochondrial volume, elevated PV levels reduced transcript levels of mitochondrial transcription factors involved in mitochondria biogenesis. In differentiated PV-overexpressing CG4 cells with a decreased mitochondrial volume, UV-induced ROS production was lower than in control CG4 cells hinting towards a possible role of PV in counteracting oxidative stress. Unexpectedly, PV also decreased the length of processes in undifferentiated CG4 cells and moreover diminished branching of differentiated CG4 cell processes, strongly correlated with the decreased density of mitochondria in CG4 cell processes. Thus besides conferring a protective role against oxidative stress, PV in a cell autonomous fashion additionally affects process’ growth and branching in CG4 cells.

Late effect of developmental exposure to glycidol on hippocampal neurogenesis in mice: Loss of parvalbumin-expressing interneurons

Developmental exposure to glycidol of rats causes axonal injury targeting axon terminals in dams and transient disruption of late-stage differentiation of hippocampal neurogenesis, accompanying sustained increase in the number of reelin-producing or calretinin-expressing interneurons in offspring. The molecular mechanism of disruptive neurogenesis probably targets the newly generating nerve terminals. We previously found differences between mice and rats in the effects on hippocampal neurogenesis after developmental exposure to the same neurotoxic substances. In the present study, we examined the effects and underlying mechanisms of developmental exposure to glycidol on hippocampal neurogenesis in mice. Glycidol (800 or 1600ppm) was administered in drinking water to mated female mice from gestational day 6 to postnatal day 21. Compared to mice drinking water without glycidol (control), the exposed dams showed axon terminal injury at both concentrations of glycidol. The offspring of the dams that had received 1600ppm glycidol had fewer parvalbumin (PVALB)+ γ-aminobutyric acid (GABA)-ergic interneurons and neuron-specific nuclear protein+ postmitotic neurons in the hilus of the hippocampal dentate gyrus. Thus, exposure of glycidol to adult mice induced axonal degeneration equivalent to that seen in the rat; however, the target mechanism for the disruption of hippocampal neurogenesis by developmental exposure was different from that in rats, with the hilar neuronal population not affected until adulthood. Considering the role of PVALB+ GABAergic interneurons in the brain, developmental glycidol exposure in mice may cause a decline in cognitive function in later life, and involve a different mechanism from that targeting axon terminals in rats.

Parvalbumin is overexpressed in the late phase of pharmacological preconditioning in skeletal muscle

Pharmacological preconditioning (PPC) with mitochondrial ATP-sensitive K(+) channel openers such as diazoxide, provides protection against ischemia in cardiac muscle, skeletal muscle, and other tissues. Effects on Ca(2+) homeostasis during the late phase of PPC have been described in cardiomyocytes, but no information is available regarding intracellular Ca(2+) changes in skeletal muscle fibers during late PPC. Intracellular Ca(2+) signals were measured in single fibers of adult mouse skeletal muscle, with fluorescent probes, 48 h after the administration of diazoxide. Parvalbumin levels in the myofibers were quantitated by Western blot. Diazoxide induction of late PPC was confirmed by partial protection of muscles from peroxide-induced damage. Late PPC was associated with a significant decrease in the duration of Ca(2+) signals during single twitches and tetanus with no changes in peak values. This effect was prevented by the reactive oxygen species (ROS) scavenger tiron. Late PPC was accompanied by a 30% increase in parvalbumin levels, and this effect was also blocked by tiron. Our data show, for the first time, a role of parvalbumin in late PPC in skeletal muscle.

Expression changes of parvalbumin and microtubule-associated protein 2 induced by chronic constriction injury in rat dorsal root ganglia.

Parvalbumin (PV), as a mobile endogenous calcium buffer, plays an important role in affecting temporospatial characteristics of calcium transients and in modulating calcium homeostasis. PV is expressed in neurons in the dorsal root ganglion (DRG) and spinal dorsal horn and may be involved in synaptic transmission through regulating cytoplasm calcium concentrations. But the exact role of PV in peripheral sensory neurons remains unknown. Microtubule-associated protein 2 (MAP-2), belonging to structural microtubule-associated protein family, is especially vulnerable to acute central nervous system (CNS) injury, and there will be rapid loss of MAP-2 at the injury site. The present study investigated the changes of PV expressing neurons and the MAP-2 neurons in the DRG after an operation for chronic constriction injury to the unilateral sciatic nerve (CCI-SN), in order to demonstrate the possible roles of PV and MAP-2 in transmission and modulation of peripheral nociceptive information. Seventy-two adult male Sprague-Dawley (SD) rats, weighing 180 - 220 g, were randomly divided into two groups (36 rats in each group), the sham operation group and chronic constriction injury (CCI) group. Six rats in each group were randomly selected to receive mechanical and thermal sensitivity tests at one day before operation and 1, 3, 5, 7, and 14 days after surgery. After pain behavioral test, ipsilateral lumbar fifth DRGs were removed and double immunofluorescence staining was performed to assess the expression changes of PV and of MAP2 expressing neurons in the L5 DRG before or after surgery. The animals with CCI-SN showed obvious mechanical allodynia and thermal hyperalgesia (P < 0.05). Both the thermal and mechanical hyperalgesia decreased to their lowest degree at 7 days after surgery compared to the baseline before surgery (P < 0.01). In normal rats before surgery, a large number of neurons were MAP-2 single labeled cells, and just a small number of PV-expressed neurons were found. PV-positive neurons, PV-positive nerve fibers and PV-negative neurons, formed a direct or close contact for cross-talk. We used immunocytochemical staining to quantify the time course of changes to PV and MAP-2 expressing neurons in tissue, and found that the number of PV expressing neurons began to slightly decrease at 3 days after surgery, and had a significant reduction at CCI day 5, day 7 (P < 0.05). But MAP-2 neurons significantly decreased on just the 3rd day after CCI (P < 0.05). No changes in PV and MAP-2 expression were almost found in sham operated rats. The number of PV positive neurons, was positively correlated with the hyperalgesia threshold. A sharp decline in MAP-2 neurons may be the early response to surgical injury, and PV positive neurons were much more effective at affecting the changes of pain behaviors, indicating that the down-regulation of PV protein could participate in, at least in part, the modulation of nociceptive transmission.

Compensatory Regulation of Cav2.1 Ca2+Channels in Cerebellar Purkinje Neurons Lacking Parvalbumin and Calbindin D-28k

Ca(v)2.1 channels regulate Ca(2+) signaling and excitability of cerebellar Purkinje neurons. These channels undergo a dual feedback regulation by incoming Ca(2+) ions, Ca(2+)-dependent facilitation and inactivation. Endogenous Ca(2+)-buffering proteins, such as parvalbumin (PV) and calbindin D-28k (CB), are highly expressed in Purkinje neurons and therefore may influence Ca(v)2.1 regulation by Ca(2+). To test this, we compared Ca(v)2.1 properties in dissociated Purkinje neurons from wild-type (WT) mice and those lacking both PV and CB (PV/CB(-/-)). Unexpectedly, P-type currents in WT and PV/CB(-/-) neurons differed in a way that was inconsistent with a role of PV and CB in acute modulation of Ca(2+) feedback to Ca(v)2.1. Ca(v)2.1 currents in PV/CB(-/-) neurons exhibited increased voltage-dependent inactivation, which could be traced to decreased expression of the auxiliary Ca(v)beta(2a) subunit compared with WT neurons. Although Ca(v)2.1 channels are required for normal pacemaking of Purkinje neurons, spontaneous action potentials were not different in WT and PV/CB(-/-) neurons. Increased inactivation due to molecular switching of Ca(v)2.1 beta-subunits may preserve normal activity-dependent Ca(2+) signals in the absence of Ca(2+)-buffering proteins in PV/CB(-/-) Purkinje neurons.

Parvalbumin: a key protein in early distal tubule NaCl reabsorption

The cortical distalnephron is committed to the fine regulation of electrolytes and water balance. Several investigations have addressed the molecular mechanisms implicated in this process. The paper by Belge et al. [1] demonstrates the emerging role of parvalbumin (PV) on distal tubule NaCl reabsorption. PV is a divalent cation buffering protein, exclusively expressed in the early distal convoluted tubule (DCT1). The authors show solid data suggesting a functional relationship between PV and the thiazide-sensitive Na+-Cl− cotransporter (NCC), the main entry step for Na+ and Cl− through the apical membrane at this nephron site. PV−/− mice exhibit a salt-losing phenotype characterized by increased diuresis, kaliuresis and high aldosterone levels, a phenotype very similar, although not identical, to the NCC knock-out mice. Accordingly, PV−/− mice manifest decreased expression of NCC, paralleled by no significant diuretic response to hydrochlorothiazide. Furthermore, in vitro studies performed on DCT cells show that PV may regulate NCC expression by modulating the Ca2+-dependent signalling pathway.

ALTERED CONTRACTILE FUNCTION IN SLOW-TWITCH MUSCLE FROM TRANSGENIC MICE OVEREXPRESSING PARVALBUMIN

827 The purpose of this study was to determine if overexpression of the Ca2+ binding protein, parvalbumin (PV), would lead to alterations in muscle contractile function and induce changes in muscle fiber-type. Two independent lines of transgenic (Tg) mice were generated with a rat PV cDNA driven by the human troponin I slow promoter which directed overexpression of PV specifically in slow (type I) fibers. In non-transgenic wild-type (Wt) littermates, PV expression in slow-twitch soleus muscle was 5% of that in the fast-twitch plantaris muscle. In Tg mice, PV expression in soleus was increased to 40-60% of that in plantaris muscle. Maximum twitch and tetanic tensions did not differ in soleus from Wt vs. Tg (n=8) but there was a rightward shift of the force-frequency curve for Tg soleus. Time to peak tension (TPT) and half-relaxation time (1/2 RT) were significantly decreased in soleus from Tg vs. Wt mice (TPT: 31 ± 1 vs. 41 ± 2 ms, p<0.05; 1/2 RT: 35 ± 2 vs. 45 ± 3 ms, p<0.05, respectively; n=4). The reduction in half-relaxation time is consistent with the role of PV in buffering cytosolic Ca2+ during muscle relaxation. Muscle fiber type, as assessed by myosin ATPase staining, was not different in Tg vs. Wt soleus suggesting that myosin heavy chain expression was not altered. There were no major changes in the expression of myoglobin, GAPDH and phospholamban assessed by reverse transcriptase polymerase chain reaction and normalized to actin expression in Tg vs. Wt soleus. These data show changes in contractile function due to alterations in Ca2+ handling without changes in myosin heavy chain or other selected markers of fiber-type specific gene expression.

Chapter 3 Neurochemical microcircuitry underlying visual and oculomotor function in the cat superior colliculus

The cat superior colliculus (SC) plays an important role in visual and oculomotor functions, including the initiation of saccadic eye movements. We have studied the organization of neurochemical specific circuits in SC that underly these functions. In this chapter we have reviewed three microcircuits that can be identified by cell type, chemical content, and synaptic input from specific afferents. The first is located within the upper sgl and is related to the W retinal pathway to this region of SC. This circuit includes relay and interneurons that contain the calcium binding protein calbindin (CB), GABA containing presynaptic dendrites, and retinal terminals that have a distribution and size typical of W retinal terminals in the cat SC. This circuit is a typical synaptic triad that mediates feedforward inhibition, possibly to regulate outflow of the W pathway to the lateral geniculate nucleus. CB neurons in SC and other structures may be uniquely related to low threshold calcium currents in these neurons. The second microcircuit consists of neurons that contain parvalbumin (PV), another calcium binding protein. These neurons are located in a dense tier with the deep sgl and upper ol and they receive input from retinal terminals that are likely from 'Y' retinal ganglion cells. Some of these neurons also project to the lateral posterior nucleus and some colocalize glutamate. We speculate that these neurons also receive cortical 'Y' input although we have yet to prove this experimentally. The role of PV in these cells is unknown, but PV has been shown to be contained in fast spiking, non-accomodating neurons in visual cortex which have very rapid spike discharges that are also characteristic of SC neurons innervated by 'Y' input. The third microcircuit consists of a group of clustered neurons within the igl of the cat SC that overlaps the patch-like innervation of afferents to this region that come from the pedunculopontine tegmental and lateral dorsal tegmental nuclie, the substantia nigra, and the cortical frontal eye fields. These clustered neurons project through the tectopontobulbar pathway and terminate within the cuneiform region (CFR) of the midbrain tegmentum. They transiently express NOS during development. Ongoing studies in our laboratory suggest that these cells receive synaptic inputs directly from the PPTN and SN and may represent functional modules involved in the initiation of saccadic eye movements.

Parvalbumin, labile heat and slowing of relaxation in mouse soleus and extensor digitorum longus muscles.

1. Parvalbumin content, heat rate and rate of relaxation were measured in two mouse muscles: the slow-twitch soleus and the fast-twitch extensor digitorum longus (EDL). 2. No trace of parvalbumin was found in the soleus; EDL contained a mean of 4.86 mg of this protein per gram of fresh muscle (S.D. = 1.25). 3. Heat rate during 7 s isometric tetani in isolated soleus muscle at 20 degrees C can be described by the sum of an exponentially decaying term and a constant term. The exponential term is reduced by 67% in a second tetanus performed 1 s after a first one; its repriming is complete after a resting period of about 1 min. The exponential term has therefore the properties of labile heat. 4. Relaxation rate measured during 15 s of isometric interrupted tetani at 20 degrees C is nearly constant in the soleus, but decreases continuously with increasing tetanus duration in the EDL. In the latter, isometric tension also decreases continuously. 5. Therefore, parvalbumin can account neither for the labile heat production in mouse soleus nor for the slowing of relaxation associated with muscle fatigue observed after a few seconds of tetanus in EDL. The role of parvalbumin in striated muscles is thus reassessed, and other possible causes of labile heat production and slowing of relaxation are discussed.

Influence of temperature on mechanics and energetics of muscle contraction

Results gleaned from use of temperature as a probe to study skeletal muscle performance and mechanisms of activation and contraction are reviewed. Steady-state and non-steady-state responses to changes in temperature are considered. Temperature sensitivities, Q10 values, of mechanical and energetic parameters range from nearly 1 to greater than 5 in frog skeletal muscle. Factors that are less temperature sensitive (Q10 less than or equal to 1.5) are peak tetanic force, instantaneous stiffness, curvature of force-velocity relation, magnitude of labile heat, and mechanical efficiency. Rates with intermediate temperature sensitivities (Q10 greater than 2 but less than 3) include rate of isometric force development, maximum shortening velocity, and relaxation from a brief tetanus. Rates with high temperature sensitivities (Q10 greater than 3) include cross-bridge turnover during an isometric tetanus, isometric economy, maximum power output, Ca2+ sequestration by sarcoplasmic reticulum, relaxation from a prolonged tetanus, and recovery metabolism. The observation that the Q10 for relaxation rate depends on tetanic duration can be explained in terms of the possible role of parvalbumin as a soluble relaxing factor.

The effects of temperature on relaxation in frog skeletal muscle: the role of parvalbumin.

Isolated muscle fibers from Rana temporaria tibialis anterior muscles were microinjected with aequorin. The force responses and the Ca2+ transients associated with twitch and tetanic contractions were studied at several temperatures. The declines of the Ca2+ transients were well described by single exponential equations and the effects of temperature were complex (multi-exponential). To determine if these temperature effects on the Ca2+ transients were influenced by the Ca2+ indicator itself, samples of the injected aequorin were studied in vitro using a Gibson stopped-flow apparatus. The quenching of aequorin luminescence with either EGTA or de-calcified Rana temporaria parvalbumin were mono-exponential. These overall quenching reactions had single exponential temperature dependencies. The effects of temperature on the declines of the single fiber Ca2+ transients did not appear to be influenced by the kinetics of the aequorin reaction. The disparity in the effects of temperature on the single fiber Ca2+ transients versus the in vitro quenching of aequorin luminescence with parvalbumin, were interpreted to indicate that in twitch and tetanic contractions of these fibers, it was unlikely that soluble Ca2+ binding proteins played a major role in the regulation of myoplasmic Ca2+.