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10.1126/science.288.5463.143
62,577,968
Atomic force microscopy and single-molecule force spectroscopy were combined to image and manipulate purple membrane patches from Halobacterium salinarum . Individual bacteriorhodopsin molecules were first localized and then extracted from the membrane; the remaining vacancies were imaged again. Anchoring forces betwee...
10.1038/nrmicro905
References 49 and 50 are outstanding papers showing how the combination of AFM imaging and single-molecule force spectroscopy allows researchers to subject membrane proteins to controlled manipulation, thereby providing details of their unfolding pathways and of the forces that anchor them to the membrane.
10.1099/mic.0.26431-0
66,244,564
Aggregation of microbial cells mediated by specific interactions plays a pivotal role in the natural environment, in medicine and in biotechnological processes. Here we used atomic force microscopy (AFM) to measure individual lectin–carbohydrate interactions involved in the flocculation of yeast cells, an aggregation e...
10.1038/nrmicro905
Biologically modified tips are used to measure the receptor–ligand interaction forces that mediate the aggregation of yeast cells.
10.1126/science.1073733
20,048,126
Constitutive Hedgehog (Hh) pathway activity is associated with initiation of neoplasia, but its role in the continued growth of established tumors is unclear. Here, we investigate the therapeutic efficacy of the Hh pathway antagonist cyclopamine in preclinical models of medulloblastoma, the most common malignant brain ...
10.1038/nrn1518
This paper was crucial in defining the role of the hedgehog pathway in the genesis of medulloblastoma, and in identifying inhibition of the hedgehog pathway as a potential therapy.
10.1146/annurev.neuro.24.1.385
42,570,933
▪ Abstract Tumors of the central nervous system (CNS) can be devastating because they often affect children, are difficult to treat, and frequently cause mental impairment or death. New insights into the causes and potential treatment of CNS tumors have come from discovering connections with genes that control cell gro...
10.1038/nrn1518
This outstanding review highlights the genetic mechanisms that are known to be involved in medulloblastoma, as well as other paediatric brain tumours.
10.1111/j.1750-3639.2002.tb00427.x
100,710,938
There are several currently employed classification systems for diffuse gliomas that sort tumors based on histological features. Contemporary molecular techniques, however, offer the promise of improved tumor classification and resultant patient stratification for treatment and prognosis. In particular, gene expression...
10.1038/nrn1518
This paper showed that cDNA-microarray technology could potentially be used to address diagnostically confusing gliomas, and that transcriptional information contains more data about outcome than does pathological examination.
10.1158/0008-5472.c.6493919.v1
136,597,145
<div>Abstract<p>In current clinical practice, histology-based grading of diffuse infiltrative gliomas is the best predictor of patient survival time. Yet histology provides little insight into the underlying biology of gliomas and is limited in its ability to identify and guide new molecularly targeted ther...
10.1038/nrn1518
In this paper, the authors demonstrate that gene-expression-based grouping of malignant gliomas is a more powerful predictor of survival than pathological type, grade or age.
10.1101/gad.1153603
38,968,679
How do very diverse signaling pathways induce neural differentiation in Xenopus ? Anti-BMP (Chordin), FGF8, and IGF2 signals are integrated in the embryo via the regulation of Smad1 phosphorylation. Neural induction results from the combined inhibition of BMP receptor serine/threonine kinases and activation of receptor...
10.1038/nrn1805
Shows that Smad1 phosphorylation by MAPK, which is activated by the IGF and FGF pathways, is crucial for the neural inhibiting activity of BMP, providing the molecular basis for the crosstalk between these signalling pathways.
10.1242/dev.127.6.1209
62,660,216
We have studied the role of Bmp signaling in patterning neural tissue through the use of mutants in the zebrafish that disrupt three different components of a Bmp signaling pathway: swirl/bmp2b, snailhouse/bmp7 and somitabun/smad5. We demonstrate that Bmp signaling is essential for the establishment of the prospective ...
10.1038/nrn1805
Describes the analysis of three zebrafish mutants ( swr snh and sbn ) that have defective BMP signalling and shows that BMP signalling is essential for spinal cord patterning and neural crest induction. The authors also show that cell fates in the spinal cord are determined by a BMP gradient.
10.1242/dev.129.10.2459
125,328,947
In the spinal neural tube, populations of neuronal precursors that express a unique combination of transcription factors give rise to specific classes of neurons at precise locations along the dorsoventral axis. Understanding the patterning mechanisms that generate restricted gene expression along the dorsoventral axis...
10.1038/nrn1805
Provides in vivo evidence that BMP signalling is essential for patterning the dorsal and intermediate spinal cord in the chick embryo.
10.2337/diabetes.54.12.3435
80,392,370
Poly(ADP-ribose) polymerase (PARP) activation, an important factor in the pathogenesis of diabetes complications, is considered a downstream effector of oxidative-nitrosative stress. However, some recent findings suggest that it is not necessarily the case and that PARP activation may precede and contribute to free rad...
10.1038/nrn2294
Together with references 22 and 23, this paper provided a definitive elucidation of the connections between glucose, the polyol pathway and oxidative and nitrosative stress, in addition to a proposed relationship with PARP activation.
10.2337/diabetes.53.7.1851
3,468,817
This study examined the role of p38 mitogen-activated protein (MAP) kinase in transducing high glucose into deficits in nerve conduction velocity (NCV) that are characteristic of diabetic neuropathy. p38 activation and NCV were measured in streptozocin-induced diabetic rats treated with a p38 inhibitor, an aldose reduc...
10.1038/nrn2294
This study linked activation of p38 to abnormal nerve conduction and established an involvement of the polyol pathway in p38 phosphorylation.
10.1523/jneurosci.0541-05.2005
103,216,848
Na v 1.6 is the major sodium channel isoform at nodes of Ranvier in myelinated axons and, additionally, is distributed along unmyelinated C-fibers of sensory neurons. Thus, modulation of the sodium current produced by Na v 1.6 might significantly impact axonal conduction. Mitogen-activated protein kinases (MAPKs) are e...
10.1038/nrn2294
This paper provided an elegant demonstration of phosphorylation of the voltage-gated sodium channel Na v 1.6 by p38, providing an explanation for the slowing of nerve conduction.
10.1523/jneurosci.1299-04.2004
83,469,160
Here, we investigated the intracellular mechanisms that underlie the relative invulnerability of adult versus developing dorsal root ganglion (DRG) sensory neurons. In culture, adult neurons were resistant to stimuli that caused apoptosis of their neonatal counterparts. In both adult and neonatal neurons, death stimuli...
10.1038/nrn2294
This paper provided an understanding of why adult sensory neurons are resistant to apoptosis, triggered by several physiological and pathophysiological stimuli.
10.2337/diabetes.52.9.2363
109,177,068
Long-term experimental diabetes may best model the prominent and irreversible sensory deficits of chronic human diabetic polyneuropathy. Whereas irretrievable loss of sensory neurons, if present, would be an unfortunate feature of the disease, systematic unbiased counting has indicated that sensory neurons survive long...
10.1038/nrn2294
This study provided a conclusive demonstration that apoptosis does not occur in the sensory neurons of rats with long-term experimental diabetes.
10.1111/j.1460-9568.1995.tb01098.x
82,638,581
Abstract Manipulation of neurotrophic support is a developing strategy for new therapy aimed at neurodegenerative diseases. This study demonstrates reduced content and retrograde transport of endogenous nerve growth factor (NGF) in sciatic nerve of diabetic rats. There were also reductions in the diabetic rats in NGF p...
10.1038/nrn2294
This paper confirmed the earlier finding of reduced NGF support of sensory neurons in diabetic rats and demonstrated that exogenous NGF can restore normal expression of responsive genes that are defective in experimental diabetes.
10.2337/db06-1269
16,768,613
Hyperalgesia to noxious stimuli is accompanied by increased spinal cyclooxygenase (COX)-2 protein in diabetic rats. The present studies were initiated to establish causality between increased spinal COX-2 activity and hyperalgesia during diabetes and to assess the potential involvement of polyol pathway activity in the...
10.1038/nrn2294
An elegant study that showed that COX2 is induced in the spinal cord of diabetic rats and its involvement in hyperalgesia — this is the clearest demonstration of a potential spinal involvement in diabetic neuropathic pain.
10.1126/science.1139597
104,236,062
Brain processing depends on the interactions between neuronal groups. Those interactions are governed by the pattern of anatomical connections and by yet unknown mechanisms that modulate the effective strength of a given connection. We found that the mutual influence among neuronal groups depends on the phase relation ...
10.1038/nrn2979
A pivotal empirical study supporting the hypothesis that phase synchronization facilitates neuronal communication ('communication through coherence').
10.1073/pnas.0911531107
20,622,158
Recent findings indicate that the hippocampus supports not only long-term memory encoding but also plays a role in working memory (WM) maintenance of multiple items; however, the neural mechanism underlying multi-item maintenance is still unclear. Theoretical work suggests that multiple items are being maintained by ne...
10.1038/nrn2979
The first direct empirical evidence that multi-item working memory relies on cross-frequency coupling between the amplitude of high-frequency and the phase of low-frequency oscillations in the human hippocampus.
10.1126/science.1085818
100,684,581
The amygdalohippocampal circuit plays a pivotal role in Pavlovian fear memory. We simultaneously recorded electrical activity in the lateral amygdala (LA) and the CA1 area of the hippocampus in freely behaving fear-conditioned mice. Patterns of activity were related to fear behavior evoked by conditioned and indifferen...
10.1038/nrn2979
This study showed that amygdala–hippocampal coherence is increased in rodents after fear conditioning, suggesting that phase synchronization is also relevant for non-declarative long-term memory processes.
10.1073/pnas.0701826104
104,423,132
The hippocampal formation is believed to be critical for the encoding, consolidation, and retrieval of episodic memories. Yet, how these processes are supported by the anatomically diverse hippocampal networks is still unknown. To examine this issue, we tested rats in a hippocampus-dependent delayed spatial alternation...
10.1038/nrn2979
In this study, intrahippocampal phase synchronization between areas CA1 and CA3 was shown to increase during retrieval of information in a delayed spatial alternation task.
10.1523/jneurosci.4250-04.2005
104,438,998
Synchronization of neuronal activity, often associated with network oscillations, is thought to provide a means for integrating anatomically distributed processing in the brain. Neuronal processing, however, involves simultaneous oscillations in various frequency bands. The mechanisms involved in the integration of suc...
10.1038/nrn2979
One of the first studies highlighting the relevance of m:n phase–phase coupling for working memory processes.
10.1182/blood-2010-09-309161
79,706,772
Abstract Hemolytic uremic syndrome (HUS) is commonly associated with Shiga toxin (Stx)–producing Escherichia coli O157:H7 infection. This study examined patient samples for complement activation on leukocyte-platelet complexes and microparticles, as well as donor samples for Stx and lipopolysaccharide (O157LPS)–induced...
10.1038/nrneph.2017.98
The paper presents complement-coated microvesicles in the circulation during HUS, suggesting a role in inflammation and thrombogenesis.
10.1182/blood-2004-03-1095
82,409,181
Abstract Tissue factor (TF) circulates in plasma, largely on monocyte/macrophage-derived microvesicles that can bind activated platelets through a mechanism involving P-selectin glycoprotein ligand-1 (PSGL-1) on the microvesicles and P-selectin on the platelets. We found these microvesicles to be selectively enriched i...
10.1038/nrneph.2017.98
Describes the importance of tissue factor and lipid rafts on microvesicles.
10.4049/jimmunol.1301221
42,570,126
Abstract This study addressed the contribution of ADAMTS13 deficiency to complement activation in thrombotic thrombocytopenic purpura (TTP). Renal tissue and blood samples were available from 12 TTP patients. C3 and C5b-9 deposition were demonstrated in the renal cortex of two TTP patients, by immunofluorescence and im...
10.1038/nrneph.2017.98
This paper describes the release of complement-coated endothelial-derived microvesicles when plasma from patients with TTP is perfused over glomerular endothelial cells.
10.4049/jimmunol.1402470
39,855,517
Abstract Shiga toxin (Stx)-producing Escherichia coli (STEC) cause hemolytic uremic syndrome (HUS). This study investigated whether Stx2 induces hemolysis and whether complement is involved in the hemolytic process. RBCs and/or RBC-derived microvesicles from patients with STEC-HUS (n = 25) were investigated for the pre...
10.1038/nrneph.2017.98
The paper describes the involvement of red blood cell-derived microvesicles in haemolysis and inhibition by purinergic receptor inhibitors.
10.1681/asn.2016060637
81,693,989
The kinin system is activated during vasculitis and may contribute to chronic inflammation. C1-inhibitor is the main inhibitor of the kinin system. In this study, we investigated the presence of the kinin B1 receptor on endothelial microvesicles and its contribution to the inflammatory process. Compared with controls (...
10.1038/nrneph.2017.98
This paper describes the chemotactic potential of endothelial-derived microvesicles positive for both kinin receptors, B1 and B2, and IL-8, and inhibition of their release by C1 inhibitor.
10.1002/cyto.a.22795
124,556,769
Abstract The interest in extracellular vesicles (EVs) has grown exponentially over the last decade. Evolving evidence is demonstrating that these EVs are playing an important role in health and disease. They are involved in intercellular communication and have been shown to transfer proteins, lipids, and nucleic acids....
10.1038/nrneph.2017.98
A review describing and comparing methods for detection of extracellular vesicles.
10.1182/blood-2007-08-106153
81,421,237
Abstract Atypical hemolytic uremic syndrome (aHUS) may be associated with mutations in the C-terminal of factor H (FH). FH binds to platelets via the C-terminal as previously shown using a construct consisting of short consensus repeats (SCRs) 15 to 20. A total of 4 FH mutations, in SCR15 (C870R) and SCR20 (V1168E, E11...
10.1038/nrneph.2017.98
This paper describes complement-coated platelets releasing tissue factor-positive microvesicles and inhibition by addition of factor H to prevent complement activation on blood cells in atypical HUS.
10.1038/srep23498
84,424,470
Abstract The miRNAs in urinary sediment are easy to obtain, which provides a new approach to searching for non-invasive biomarkers of IgA nephropathy (IgAN). Compared with normal controls (n = 3), 214 different miRNAs in the urinary sediment of IgAN (n = 9) were found by miRNA chip assay. By quantitative PCR analysis, ...
10.1038/nrneph.2017.98
A paper describing urinary erythrocyte miRNAs derived from microvesicles as biomarkers of IgA nephropathy.
10.1128/mcb.00240-10
62,350,612
ABSTRACT Recruitment to telomeres is a pivotal step in the function and regulation of human telomerase; however, the molecular basis for recruitment is not known. Here, we have directly investigated the process of telomerase recruitment via fluorescence in situ hybridization (FISH) and chromatin immunoprecipitation (Ch...
10.1038/nsmb.3083
This work thoroughly investigates the shelterin requirements for telomerase recruitment to telomeres
10.1126/science.1165357
20,832,138
Telomerase is a ribonucleoprotein (RNP) complex that synthesizes telomere repeats in tissue progenitor cells and cancer cells. Active human telomerase consists of at least three principal subunits, including the telomerase reverse transcriptase, the telomerase RNA (TERC), and dyskerin. Here, we identify a holoenzyme su...
10.1038/nsmb.3083
Refs. 24 and 25 report the discovery of the protein TCAB1 (WDR79) and its association with an RNA motif for RNP CB localization
10.1101/gad.246819.114
83,479,467
Telomere length homeostasis is essential for the long-term survival of stem cells, and its set point determines the proliferative capacity of differentiated cell lineages by restricting the reservoir of telomeric repeats. Knockdown and overexpression studies in human tumor cells showed that the shelterin subunit TPP1 r...
10.1038/nsmb.3083
This work, through genome editing, investigates the functions of TPP1 in human pluripotent stem cells and uncovers a multiplicity of TPP1 requirements for telomerase recruitment and activation
10.1073/pnas.0808953105
125,107,058
Calcium oscillations suppress mitochondrial movements along the microtubules to support on-demand distribution of mitochondria. To activate this mechanism, Ca 2+ targets a yet unidentified cytoplasmic factor that does not seem to be a microtubular motor or a kinase/phosphatase. Here, we have studied the dependence of m...
10.1038/nrn3156
References 23, 24 and 25 independently identified MIRO as a Ca 2+ sensor, providing a mechanism for the underlying Ca 2+ -dependent regulation of mitochondrial mobility.
10.1091/mbc.e05-06-0526
107,178,976
To address questions about mechanisms of filament-based organelle transport, a system was developed to image and track mitochondria in an intact Drosophila nervous system. Mutant analyses suggest that the primary motors for mitochondrial movement in larval motor axons are kinesin-1 (anterograde) and cytoplasmic dynein ...
10.1038/nrn3156
This study provides the genetic evidence in the D. melanogaster nervous system that dynein motors have a crucial role in mitochondrial retrograde transport in axons.
10.1083/jcb.114.2.295
16,660,194
Biochemical, pharmacological and immunocytochemical studies have implicated the microtubule-activated ATPase, kinesin, in the movement of membrane bounded organelles in fast axonal transport. In vitro studies suggested that kinesin moves organelles preferentially in the anterograde direction, but data about the functio...
10.1038/nrn3156
This study shows that KIF5 motors are attached to brain mitochondria.
10.1083/jcb.200506042
83,358,556
In neurons, proper distribution of mitochondria in axons and at synapses is critical for neurotransmission, synaptic plasticity, and axonal outgrowth. However, mechanisms underlying mitochondrial trafficking throughout the long neuronal processes have remained elusive. Here, we report that syntabulin plays a critical r...
10.1038/nrn3156
This study identifies syntabulin as a second prominent KIF5 motor adaptor for mitochondria. Syntabulin loss-of-function or interference of the syntaphilin–KIF5 interaction reduces anterograde, but not retrograde, mitochondrial transport along axons.
10.1083/jcb.94.1.129
62,621,353
The elaborate cross-connections among membranous organelles (MO), microtubules (MT), and neurofilaments (NF) were demonstrated in unifixed axons by the quick-freeze, deep-etch, and rotary-shadowing method. They were categorized into three groups: NF-associated cross-linker, MT-associated cross-bridges, and long cross-l...
10.1038/nrn3156
This study provides the first morphological evidence for the crossbridges between axonal mitochondria and microtubules or neurofilaments.
10.1242/jcs.01130
104,401,960
Disruption of axonal transport leads to a disorganized distribution of mitochondria and other organelles and is thought to be responsible for some types of neuronal disease. The reason for bidirectional transport of mitochondria is unknown. We have developed and applied a set of statistical methods and found that axona...
10.1038/nrn3156
This study demonstrates that mitochondria with a high membrane potential are transported anterogradely towards distal processes, whereas damaged mitochondria return to the cell body following acute depolarization.
10.1128/mmbr.62.1.181-203.1998
19,488,675
SUMMARY To withstand the high intracellular pressure, the cell wall of most bacteria is stabilized by a unique cross-linked biopolymer called murein or peptidoglycan. It is made of glycan strands [poly-(GlcNAc-MurNAc)], which are linked by short peptides to form a covalently closed net. Completely surrounding the cell,...
10.1038/nrmicro2677
A landmark review on peptidoglycan synthesis in E. coli , with details of the '3 for 1' growth model.
10.1111/j.1365-2958.2006.05280.x
100,342,043
Summary The murein (peptidoglycan) sacculus is an essential polymer embedded in the bacterial envelope. The Escherichia coli class B penicillin‐binding protein (PBP) 3 is a murein transpeptidase and essential for cell division. In an affinity chromatography experiment, the bifunctional transglycosylase‐transpeptidase m...
10.1038/nrmicro2677
The first study to provide evidence for a direct interaction between peptidoglycan synthases.
10.1073/pnas.0904030106
81,689,931
Drug-resistant bacteria have caused serious medical problems in recent years, and the need for new antibacterial agents is undisputed. Transglycosylase, a multidomain membrane protein essential for cell wall synthesis, is an excellent target for the development of new antibiotics. Here, we determined the X-ray crystal ...
10.1038/nrmicro2677
This article describes the crystal structure of PBP1B, which includes the transmembrane region, and suggests a model for the coupling of GTase and TPase reactions.
10.1073/pnas.0808035105
60,447,245
The stress-bearing component of the bacterial cell wall—a multi-gigadalton bag-like molecule called the sacculus—is synthesized from peptidoglycan. Whereas the chemical composition and the 3-dimensional structure of the peptidoglycan subunit (in at least one conformation) are known, the architecture of the assembled sa...
10.1038/nrmicro2677
In this study, ECT solves a long-standing dispute about the orientation of the glycan chains in the single-layered peptidoglycan in Gram-negative bacteria.
10.1073/pnas.1108999108
61,055,557
Bacterial cells possess multiple cytoskeletal proteins involved in a wide range of cellular processes. These cytoskeletal proteins are dynamic, but the driving forces and cellular functions of these dynamics remain poorly understood. Eukaryotic cytoskeletal dynamics are often driven by motor proteins, but in bacteria n...
10.1038/nrmicro2677
References 47–49 describe high-resolution fluorescence microscopy showing peptidoglycan synthesis-dependent movement of MreB perpendicular to the long axis.
10.1111/j.1365-2958.2007.05720.x
82,540,449
Summary The tubulin homologue FtsZ is well known for its essential function in bacterial cell division. Here, we show that in Caulobacter crescentus , FtsZ also plays a major role in cell elongation by spatially regulating the location of MurG, which produces the essential lipid II peptidoglycan cell wall precursor. Th...
10.1038/nrmicro2677
This investigation demonstrates the FtsZ-dependent preseptal phase of cell elongation in C. crescentus
10.1101/gad.1878110
109,038,873
How proteins catalyze morphogenesis is an outstanding question in developmental biology. In bacteria, morphogenesis is intimately linked to remodeling the cell wall exoskeleton. Here, we investigate the mechanisms by which the mother cell engulfs the prospective spore during sporulation in Bacillus subtilis . A membran...
10.1038/nrmicro2677
This and reference 93 show for the first time that septum-splitting peptidoglycan hydrolases require activation by other proteins.
10.1073/pnas.93.16.8733
79,469,063
We applied the directional tag PCR subtractive hybridization method to construct a rat hypothalamic cDNA library from which cerebellar and hippocampal sequences had been depleted, enriching 20-30-fold for sequences expressed selectively in the hypothalamus. We studied a sample of 94 clones selected for enrichment in th...
10.1038/nrn808
The first detection of the mRNA that encodes preprohypocretin in a few thousand neurons in the lateral hypothalamus.
10.1073/pnas.95.1.322
20,687,273
We describe a hypothalamus-specific mRNA that encodes preprohypocretin, the putative precursor of a pair of peptides that share substantial amino acid identities with the gut hormone secretin. The hypocretin (Hcrt) protein products are restricted to neuronal cell bodies of the dorsal and lateral hypothalamic areas. The...
10.1038/nrn808
The prediction of two carboxy-terminally amidated peptides, immunohistochemical mapping of the projections of the hypocretin neurons, detection of the peptides in dense-core vesicles at synapses and demonstration that the synthetic, amidated peptides have excitatory electrophysiological activity.
10.1073/pnas.96.19.10911
80,240,697
The localization of orexin neuropeptides in the lateral hypothalamus has focused interest on their role in ingestion. The orexigenic neurones in the lateral hypothalamus, however, project widely in the brain, and thus the physiological role of orexins is likely to be complex. Here we describe an investigation of the ac...
10.1038/nrn808
A detailed characterization of the endocrine regulatory effects of the hypocretins and, with reference 108 , the first direct experimental evidence that the hypocretins influence arousal by exciting locus coeruleus neurons.
10.1126/science.4023713
80,219,712
Single cells were recorded in the visual cortex of monkeys trained to attend to stimuli at one location in the visual field and ignore stimuli at another. When both locations were within the receptive field of a cell in prestriate area V4 or the inferior temporal cortex, the response to the unattended stimulus was dram...
10.1038/nrn3443
On the basis of the finding that attention biases the response to two stimuli inside a neuron's RF in favour of the attended stimulus, the authors are the first to suggest that attention alters spatial integration by contracting RFs around the attended stimulus.
10.1152/jn.1996.75.3.1306
40,518,317
1. We studied the spatial interaction between stimulus and attention in macaque area V4. Monkeys were required to fixate a small spot while continuously attending to a ring-shaped target within a large array of identical rings. Meanwhile, the response of the V4 cell under study was tested by flashing behaviorally irrel...
10.1038/nrn3443
This study provides the first evidence for shifts of a neuron's RF profile towards an attended stimulus.
10.1046/j.1365-2958.2003.03428.x
18,509,670
Summary The torCAD operon encoding the TMAO reductase respiratory system is induced in the presence of TMAO by the two‐component regulatory system TorS/TorR. The TorS sensor detects TMAO and transphosphorylates the TorR response regulator via a four‐step phosphorelay. Once phosphorylated, TorR activates expression of t...
10.1038/nrmicro2549
An illustration of a TCS that senses and pre-emptively responds to a metabolic change that will lead to a subsequent alkaline challenge to E. coli growth.
10.1073/pnas.0510914103
79,413,360
Sodium proton antiporters are essential enzymes that catalyze the exchange of sodium ions for protons across biological membranes. Protonations and deprotonations of individual amino acid residues and of clusters formed by these residues play an important role in activating these enzymes and in the mechanism of transpo...
10.1038/nrmicro2549
Crystal structure-based computation reveals clusters of tightly electrostatically interacting residues in a transmembrane region of NhaA, and shows that these clusters have functional importance in pH regulation of the antiporter.
10.1126/science.287.5452.482
62,514,994
Acidic media trigger cytoplasmic urease activity of the unique human gastric pathogen Helicobacter pylori. Deletion of ureI prevents this activation of cytoplasmic urease that is essential for bacterial acid resistance. UreI is an inner membrane protein with six transmembrane segments as shown by in vitro transcription...
10.1038/nrmicro2549
This is the first paper to identify a proton-gated urea channel in bacteria: the UreI channel, which has a central role in acid acclimation of H. pylori
10.1128/iai.63.5.1669-1673.1995
123,563,121
The aim of this work was to study the significance of the urease enzyme in promoting Helicobacter pylori survival in various environments. A urease-positive H. pylori isolate, strain N6, and an isogenic urease-negative strain, strain N6(ureB::TnKm), were incubated in phosphate-buffered saline at a pH ranging from 2.2 t...
10.1038/nrmicro2549
This article describes the important observation that H. pylori does not survive in the presence of urea under non-acidic conditions, an observation that led to the unravelling of the downregulation of acid-response elements at high pH.
10.1042/bj1780195
101,048,807
Thiobacillus ferro-oxidans is capable of using the oxidation of Fe2+ by O2 at pH 2.0 as the sole source of energy for growth and CO2 fixation. The bacterium maintains an intracellular pH of 6.5 over a range of external pH from 1.0 to 8.0, as measured by [14C]acetate and [3H]methylamine distribution. The membrane potent...
10.1038/nrmicro2549
A demonstration of the PMF parameters for an extreme acidophile.
10.1111/j.1365-2958.1994.tb01329.x
45,538,924
Summary An alkali‐sensitive mutant, 38154, of the alkalophilic Bacillus sp. strain C‐125 could not grow at an alkaline pH. The nucleotide sequence of a 3.7 kb parental DNA fragment that recovers the growth of 38154 at alkaline pH has four open reading frames (ORF1–4). By sub‐cloning the fragment, we demonstrated that a...
10.1038/nrmicro2549
This investigation of non-alkaliphilic, antiporter-negative mutants results in the identification of genes encoding part of an Mrp-type antiporter that is central to pH homeostasis of alkaliphilic Bacillus spp.
10.1128/jb.176.11.3111-3116.1994
62,218,605
The effect of external pH on growth of alkaliphilic Bacillus firmus OF4 was studied in steady-state, pH-controlled cultures at various pH values. Generation times of 54 and 38 min were observed at external pH values of 7.5 and 10.6, respectively. At more alkaline pH values, generation times increased, reaching 690 min ...
10.1038/nrmicro2549
This study of PMF parameters, cytoplasmic pH and growth rates at pH values from pH 7.5 to pH >11.0 demonstrates the robust pH homeostatic capacity of alkaliphilic B. pseudofirmus OF4 and its ability to grow, after exceeding that capacity, with a cytoplasmic pH as high as 9.5.
10.1073/pnas.0402692101
122,498,580
The prokaryotic voltage-gated Na + channel, NaChBac, is one of a growing channel superfamily of unknown function. Here we show that Na V BP, the NaChBac homologue encoded by ncbA in alkaliphilic Bacillus pseudofirmus OF4, is a voltage-gated Na + channel potentiated by alkaline pH. Na V BP has roles in motility, chemota...
10.1038/nrmicro2549
This study reveals the role of a bacterial voltage-gated Na + channel in supporting the re-entry of Na + to support Na + /H + antiport-dependent pH homeostasis in an alkaliphilic Bacillus species.
10.1073/pnas.0702300104
58,368,771
Helicobacter pylori is a gastric-dwelling pathogen responsible, with acid secretion, for peptic ulcer and a 20-fold increase in the risk of gastric cancer. Several transcriptomes have been described after short-term exposure to acidity in vitro , but there are no data identifying the effects of chronic gastric exposure...
10.1038/nrmicro2549
The first paper to elucidate the in vivo transcriptome of H. pylori , supporting indications that the natural environment of this gastric pathogen is acidic with the finding that many acid acclimation genes have increased expression when compared with their expression in vitro at neutral pH.
10.1126/science.6801762
20,856,776
After infection and a prolonged incubation period, the scrapie agent causes a degenerative disease of the central nervous system in sheep and goats. Six lines of evidence including sensitivity to proteases demonstrate that this agent contains a protein that is required for infectivity. Although the scrapie agent is irr...
10.1038/nrn1584
The development of the protein-only hypothesis into biochemical terms, and the original definition of the term prion to differentiate these agents from conventional pathogens.
10.1126/science.283.5409.1935
86,443,875
Prion propagation involves the conversion of cellular prion protein (PrP C ) into a disease-specific isomer, PrP Sc , shifting from a predominantly α-helical to β-sheet structure. Here, conditions were established in which recombinant human PrP could switch between the native α conformation, characteristic of PrP C , a...
10.1038/nrn1584
This paper shows that recombinant PrP can be interconverted between native and fibrilogenic conformations that are rich in α- and β-secondary structure, respectively, under reducing and acidic conditions that are comparable to those found in the endocytic compartment.
10.1073/pnas.97.18.10248
20,254,209
Transmission of prions between mammalian species is thought to be limited by a “species barrier,” which depends on differences in the primary structure of prion proteins in the infecting inoculum and the host. Here we demonstrate that a strain of hamster prions thought to be nonpathogenic for conventional mice leads to...
10.1038/nrn1584
This article reports on the uncoupling of neurotoxicity and PrP Sc (and prion) accumulation in vivo , and describes sub-clinical prion infection in an animal model, in which animals do not develop clinical scrapie during their lifespan despite high prion titres in the brain. This paper also challenges current concepts ...
10.1126/science.1090187
58,565,697
The mechanisms involved in prion neurotoxicity are unclear, and therapies preventing accumulation of PrP Sc , the disease-associated form of prion protein (PrP), do not significantly prolong survival in mice with central nervous system prion infection. We found that depleting endogenous neuronal PrP c in mice with esta...
10.1038/nrn1584
The first report of an intervention that halts the progression of established CNS prion disease and reverses early spongiform pathology. This interruption of onset of clinical disease occurred despite the continued accumulation of PrP Sc and a rise in prion titre to levels seen in end-stage conventional infections.
10.1126/science.1073725
62,565,592
Changes in prion protein (PrP) folding are associated with fatal neurodegenerative disorders, but the neurotoxic species is unknown. Like other proteins that traffic through the endoplasmic reticulum, misfolded PrP is retrograde transported to the cytosol for degradation by proteasomes. Accumulation of even small amoun...
10.1038/nrn1584
A study arguing that aberrant cellular processing and the cytoplasmic location of PrP are mechanisms of prion neurotoxicity, which is challenged by others (see references 77 and 78).
10.1126/science.279.5352.827
20,784,195
At the endoplasmic reticulum membrane, the prion protein (PrP) can be synthesized in several topological forms. The role of these different forms was explored with transgenic mice expressing PrP mutations that alter the relative ratios of the topological forms. Expression of a particular transmembrane form (termed Ctm ...
10.1038/nrn1584
A paper that describes the transmembrane forms of particular PrP mutants and their associated cellular toxicity, consistent with the neurotoxicity of altered PrP topography in some cases.
10.1126/science.1094273
109,135,019
Neuronal death is a prominent, but poorly understood, pathological hallmark of prion disease. Notably, in the absence of the cellular prion protein (PrP C ), the disease-associated isoform, PrP Sc , appears not to be intrinsically neurotoxic, suggesting that PrP C itself may participate directly in the prion neurodegen...
10.1038/nrn1584
A report of rapid neuronal apoptosis in mice that underwent stereotaxic hippocampal inoculation with PrP antibodies, indicating a possible link between PrPC crosslinking and cell death and cell survival signalling pathways.
10.1126/science.1079469
41,472,028
Soluble oligomers are common to most amyloids and may represent the primary toxic species of amyloids, like the Aβ peptide in Alzheimer's disease (AD). Here we show that all of the soluble oligomers tested display a common conformation-dependent structure that is unique to soluble oligomers regardless of sequence. The ...
10.1038/nrn1584
A study that implicates the common structure of soluble amyloid oligomers in mechanisms of neurotoxicity in protein misfolding diseases, further developing the concept of toxic intermediate species in these disorders.
10.1073/pnas.151242598
81,152,082
Exposure of susceptible neuroblastoma N2a cells to mouse scrapie prions leads to infection, as evidenced by the continued presence of the scrapie form of the prion protein (PrP Sc ) and infectivity after 300 or more cell doublings. We find that exposure to phosphatidylinositol-specific phospholipase C (PIPLC) or to the...
10.1038/nrn1584
The first of a series of in vitro and in vivo studies that show the potential of passive immunization with anti-PrP antibodies to treat prion disease (see also references 102–106).
10.1073/pnas.121172098
100,816,319
The incubation period (IP) and the neuropathology of transmissible spongiform encephalopathies (TSEs) have been extensively used to distinguish prion isolates (or strains) inoculated into panels of inbred mouse strains. Such studies have shown that the bovine spongiform encephalopathy (BSE) agent is indistinguishable f...
10.1038/nrn1584
This article and references 9 and 140 describe important genetic linkage studies that confirm the presence of several key loci that determine the prion incubation period in mice, in addition to Prnp
10.1038/tp.2015.201
28,047,337
Abstract Shifting policies towards legalisation of cannabis for therapeutic and recreational use raise significant ethical issues for health-care providers seeking evidence-based recommendations. We investigated whether heavy cannabis use is associated with persistent harms to the hippocampus, if exposure to cannabidio...
10.1038/nrn.2016.28
This cross-sectional study used hair analysis to examine the relationship between cannabinoids and hippocampal integrity, and found that chronic Δ 9 -THC exposure is associated with reduced hippocampal volume and N -acetylaspartate concentrations; however, these effects are not found in those individuals with CBD as we...
10.1073/pnas.1516648113
123,071,838
Marijuana is one of the most commonly used drugs in the United States, and use during adolescence—when the brain is still developing—has been proposed as a cause of poorer neurocognitive outcome. Nonetheless, research on this topic is scarce and often shows conflicting results, with some studies showing detrimental eff...
10.1038/nrn.2016.28
References 163 and 164 both examined associations between adolescent cannabis use and IQ by using data from large longitudinal cohorts in the United Kingdom and United States; both papers suggest that there are non-causal explanations for the observed associations between adolescent cannabis use and lower IQ.
10.1111/j.1360-0443.2010.03139.x
124,810,481
ABSTRACT Aim To illustrate how limitations in the cannabis literature undermine our ability to understand cannabis‐related harms and problems experienced by users and identify users at increased risk of experiencing adverse outcomes of use. Method and results Limitations have been organized into three overarching theme...
10.1038/nrn.2016.28
An insightful review covering methodological inconsistencies in cannabis research, offering useful guidance for future research.
10.1038/tp.2015.219
60,005,887
Abstract Smoking cannabis daily doubles an individual’s risk of developing a psychotic disorder, yet indicators of specific vulnerability have proved largely elusive. Genetic variation is one potential risk modifier. Single-nucleotide polymorphisms in the AKT1 and catechol-O-methyltransferase (COMT) genes have been imp...
10.1038/nrn.2016.28
This is the largest study to date conducted on the acute response to cannabis and shows that psychotic-like symptoms in young healthy cannabis users are predicted by their AKT1 genotype, replicating the same effect observed in schizophrenia (see references 178 and 179).
10.1192/bjp.bp.107.046649
42,510,496
Summary Cannabis contains various cannabinoids, two of which have almost opposing actions: δ9-tetrahydrocannabinol (Δ9-THC) is psychotomimetic, whereas cannabidiol (CBD) has antipsychotic effects. Hair samples were analysed to examine levels of Δ9-THC and CBD in 140 individuals. Three clear groups emerged: ‘THC only’, ...
10.1038/nrn.2016.28
The first study to show that psychosis-like symptoms are greater in those whose hair contains only Δ 9 -THC than in those whose hair contains both Δ 9 -THC and CBD or no cannabinoids — suggesting a protective effect of CBD.
10.1126/science.1137450
60,219,766
Long-term potentiation (LTP), which approximates Hebb's postulate of associative learning, typically requires depolarization-dependent glutamate receptors of the NMDA ( N -methyl-D-aspartate) subtype. However, in some neurons, LTP depends instead on calcium-permeable AMPA-type receptors. This is paradoxical because int...
10.1038/nrn2207
This study uncovered a novel induction rule for LTP: co-incidence of presynaptic glutamate release with postsynaptic quiescence is needed. This is due to the rectification of Ca 2+ -permeable AMPA receptors, which are blocked at depolarized membrane potentials.
10.1073/pnas.161493498
19,843,871
Hippocampal inhibitory interneurons play important roles in controlling the excitability and synchronization of pyramidal cells, but whether they express long-term synaptic plasticity that contributes to hippocampal network function remains uncertain. We found that pairing postsynaptic depolarization with θ-burst stimu...
10.1038/nrn2207
This study showed that long-term potentiation can be elicited in interneurons of the stratum oriens, where it requires mGluR1.
10.1126/science.285.5432.1411
83,044,034
Long-term depression (LTD) is a well-known form of synaptic plasticity of principal neurons in the mammalian brain. Whether such changes occur in interneurons is still controversial. CA3 hippocampal interneurons expressing Ca 2+ -permeable AMPA receptors exhibited LTD after tetanic stimulation of CA3 excitatory inputs....
10.1038/nrn2207
First report to show that the induction of long-term depression requires both postsynaptic Ca 2+ , entering through Ca 2+ -permeable-AMPA receptors, and presynaptic mGluR7 activation.
10.1126/science.279.5355.1368
1,778,086
Mossy fiber synaptic transmission at hippocampal CA3 pyramidal cells and interneurons was compared in rat brain slices to determine whether mossy terminals are functionally equivalent. Tetanic stimulation of mossy fibers induced long-term potentiation in pyramidal neurons but was either without effect or it induced dep...
10.1038/nrn2207
This paper showed that the same presynaptic activity pattern in mossy fibres can induce long-term depression at synapses on interneurons and long-term potentiation at synapses on pyramidal cells.
10.1101/gad.17352611
107,269,941
Analyses of small RNA expression profiles have revealed that several DNA viruses—including particularly, herpesviruses—express high levels of multiple viral microRNAs (miRNAs) in infected cells. Here, I review our current understanding of how viral miRNAs influence viral replication and pathogenesis and discuss how vir...
10.1038/nrmicro2971
A discussion of the physiologogical interplay between viruses and miRNAs.
10.1261/rna.2303610
38,404,255
Cellular utilization of RNA interference (RNAi) as a mechanism to combat virus infection is thought to be restricted to plants and invertebrates. In vertebrates, antiviral defenses are largely dependent on interferons (IFNs), with the use of small RNAs restricted to microRNA (miRNA)–mediated targeting of host transcrip...
10.1038/nrmicro2971
This and reference 131 demonstrate that pri-miRNAs can be generated by engineered cytoplasmic viruses.
10.1261/rna.032268.112
81,613,015
microRNAs (miRNAs) represent a class of noncoding RNAs that fine-tune gene expression through post-transcriptional silencing. While miRNA biogenesis occurs in a stepwise fashion, initiated by the nuclear microprocessor, rare noncanonical miRNAs have also been identified. Here we characterize the molecular components an...
10.1038/nrmicro2971
The first article to address the mechanism enabling cytoplasmic synthesis of pri-miRNAs.
10.1111/j.1365-2990.1978.tb00545.x
26,861,835
Acetyl‐ and butyryl‐cholinesterase activities have been measured biochemically in normal brain tissue, in senile dementia of Alzheimer type and in mental disorders without Alzheimer‐type abnormalities. Acetylcholinesterase was significantly reduced and butyrylcholinesterase significantly increased, compared with the no...
10.1038/nrn1035
One of the first papers showing an increase in brain BuChE activity in AD.
10.1073/pnas.90.2.683
59,304,251
Neurofibrillary tangles and amyloid plaques express acetylcholinesterase and butyrylcholinesterase activity in Alzheimer disease. We previously reported that traditional acetylcholinesterase inhibitors such as BW284C51, tacrine, and physostigmine were more potent inhibitors of the acetylcholinesterase in normal axons a...
10.1038/nrn1035
An inhibitor-based investigation showing that the cholinesterase activity that is found in plaques and tangles in AD is aryl acylamidase activity.
10.1038/ncomms7820
118,327,009
Abstract Synaptic plasticity is the cornerstone of processes underlying persistent nociceptive activity-induced changes in normal nociceptive sensitivity. Kalirin-7 is a multifunctional guanine-nucleotide-exchange factor (GEF) for Rho GTPases that is characterized by its localization at excitatory synapses, interaction...
10.1038/nrn.2016.162
This was the first study to report intracellular signalling mechanisms recruited at spinal glutamatergic synapses that mediate synaptic potentiation and structural plasticity at nociceptive synapses in inflammatory pain.
10.1111/j.1600-0854.2011.01325.x
104,013,778
Polarized kinesin‐driven transport is crucial for development and maintenance of neuronal polarity. Kinesins are thought to recognize biochemical differences between axonal and dendritic microtubules in order to deliver their cargoes to the appropriate domain. To identify kinesins that mediate polarized transport, we p...
10.1038/nrn.2016.100
This systematic screen of kinesin motor domains shows that no motors are dendrite selective.
10.1523/jneurosci.3779-13.2014
62,046,465
The polarized distribution of membrane proteins to axonal or somatodendritic neuronal compartments is fundamental to nearly every aspect of neuronal function. The polarity of dendritic proteins depends on selective microtubule-based transport; the vesicles that carry these proteins are transported into dendrites but do...
10.1038/nrn.2016.100
This paper, which characterizes selective dendritic transport, shows that dendritically polarized vesicles reverse direction at the proximal part of the axon initial segment.
10.1073/pnas.1416544112
125,165,373
Axon initial segments (AISs) and nodes of Ranvier are sites of clustering of voltage-gated sodium channels (VGSCs) in nervous systems of jawed vertebrates that facilitate fast long-distance electrical signaling. We demonstrate that proximal axonal polarity as well as assembly of the AIS and normal morphogenesis of node...
10.1038/nrn.2016.100
This paper shows that the loss of the axon initial segment does not disrupt selective dendritic transport.
10.1126/science.6867718
62,367,876
The responses of superior colliculus cells to a given sensory stimulus were influenced by the presence or absence of other sensory cues. By pooling sensory inputs, many superior colliculus cells seem to amplify the effects of subtle environmental cues in certain conditions, whereas in others, responses to normally effe...
10.1038/nrn2331
This seminal paper demonstrated multisensory integration in single superior colliculus neurons.
10.1152/jn.2001.85.2.506
18,507,749
The majority of multisensory neurons in the cat superior colliculus (SC) are able to synthesize cross-modal cues (e.g., visual and auditory) and thereby produce responses greater than those elicited by the most effective single modality stimulus and, sometimes, greater than those predicted by the arithmetic sum of thei...
10.1038/nrn2331
The critical nature of influences from two cortical areas on multisensory integration in superior colliculus neurons is demonstrated here.
10.1152/jn.1996.76.3.2071
59,333,043
1. The lateral intraparietal area (LIP) of the posterior parietal cortex lies within the dorsal cortical stream for spatial vision and processes visual information to plan saccadic eye movements. We investigated how LIP neurons respond when a monkey makes saccades to the remembered location of sound sources in the abse...
10.1038/nrn2331
This was one of the first studies to demonstrate that the auditory receptive fields of LIP neurons can be coded in an eye-centred reference frame.
10.1523/jneurosci.3550-06.2006
45,485,673
The integration of auditory and visual stimuli is crucial for recognizing objects, communicating effectively, and navigating through our complex world. Although the frontal lobes are involved in memory, communication, and language, there has been no evidence that the integration of communication information occurs at t...
10.1038/nrn2331
This paper is one of the first to examine multisensory integration in single neurons of the prefrontal cortex. It suggests a role for this region in the integration of the visual and auditory stimuli that constitute coherent communication signals.
10.1162/089892999563544
20,805,631
The aim of this study was (1) to provide behavioral evidence for multimodal feature integration in an object recognition task in humans and (2) to characterize the processing stages and the neural structures where multisensory interactions take place. Event-related potentials (ERPs) were recorded from 30 scalp electrod...
10.1038/nrn2331
This early event-related-potential study in human subjects demonstrated that multisensory integration can take place very early in the processing of auditory and visual information.
10.1152/jn.00497.2006
79,683,054
Multisensory integration refers to the process by which the brain synthesizes information from different senses to enhance sensitivity to external events. In the present experiments, animals were reared in an altered sensory environment in which visual and auditory stimuli were temporally coupled but originated from di...
10.1038/nrn2331
This was the first paper to show that the gradual postnatal development of multisensory integration in superior colliculus neurons allows early experience to form its operational principles.
10.1242/dev.128.21.4189
83,489,348
Anteroposterior (AP) patterning of the vertebrate neural plate is initiated during gastrulation and is regulated by Spemann’s organizer and its derivatives. The prevailing model for AP patterning predicts a caudally increasing gradient of a ‘transformer’ which posteriorizes anteriorly specified neural cells. However, t...
10.1038/nrn1665
This is the first clear demonstration that a gradient of WNT signalling is responsible for the A–P patterning of the neural plate. A specific read-out of WNT activity using levels of β-catenin shows that a WNT morphogen gradient patterns the Xenopus nervous system.
10.1126/science.1089610
102,350,281
Commissural neurons in the mammalian dorsal spinal cord send axons ventrally toward the floor plate, where they cross the midline and turn anteriorly toward the brain; a gradient of chemoattractant(s) inside the spinal cord controls this turning. In rodents, several Wnt proteins stimulate the extension of commissural a...
10.1038/nrn1665
This paper provides the first evidence that WNT signalling through FZ regulates axon guidance in vertebrates. WNT4, which is expressed in an A–P gradient in the floor plate, stimulates axon outgrowth and turning of commissural axons. This effect is mediated through FZ3, as mice lacking FZ3 show a defect in the normal a...
10.1111/j.1471-4159.1990.tb01236.x
20,787,815
Abstract: To provide an “in vitro” system for studying brain capillary function, we have developed a process ofcoculture that closely mimics the “in vivo” situation by culturing brain capillary endothelial cells on one side of a filter and astrocytes on the other. Under these conditions, endothelial cells retain all th...
10.1038/nrn1824
One of the first papers to describe a reliable method for generating an endothelial–astrocyte co-culture model of the BBB tight enough for study of permeability and transport. The model has since been successful in functional and mechanistic studies.
10.1002/ana.20369
83,506,605
Abstract Parkinson's disease (PD) is associated with a loss of neurons from the midbrain. The cause of PD is unknown, but it is established that certain neurotoxins can cause similar syndromes. The brain is normally protected from these noxious blood‐borne chemicals by the blood–brain barrier which includes specialized...
10.1038/nrn1824
An important but controversial paper showing how modern imaging techniques can be used to investigate BBB transport function in humans, and the insight this may give into disease states.
10.1046/j.1471-4159.2003.01829.x
41,710,432
Abstract Evidence suggests that endothelin‐1 (ET‐1) plays an essential role in brain inflammation. However, whether ET‐1 contributes directly to blood–brain barrier (BBB) breakdown remains to be elucidated. Using an in vitro BBB model consisting of co‐cultures of human primary astrocytes and brain microvascular endothe...
10.1038/nrn1824
Illustrates the potential complexities of signalling between cells at the BBB — even apparently direct actions may involve indirect loops and potentiating (and inhibitory) modulation.
10.1152/jn.2001.85.5.1969
20,393,455
In this first intracellular study of neocortical activities during waking and sleep states, we hypothesized that synaptic activities during natural states of vigilance have a decisive impact on the observed electrophysiological properties of neurons that were previously studied under anesthesia or in brain slices. We i...
10.1038/nrn1325
The first intracellular study of four cortical neuronal types during the natural waking–sleep cycle, which analysed state-related changes in membrane potential, input resistance and incidence of various cell classes.
10.1126/science.3059497
81,580,704
This article reviews the electroresponsive properties of single neurons in the mammalian central nervous system (CNS). In some of these cells the ionic conductances responsible for their excitability also endow them with autorhythmic electrical oscillatory properties. Chemical or electrical synaptic contacts between th...
10.1038/nrn1325
A visionary review article by a pioneer of in vitro investigations on intrinsic neuronal properties in mammals, which changed our view of cellular operations in different brain structures.