Vincent Pieribone

Yale University | YU · Department of Cellular and Molecular Physiology

Monitoring spiking activity across large neuronal populations at behaviorally relevant timescales is critical for understanding neural circuit function. Unlike calcium imaging, voltage imaging requires kilohertz sampling rates that reduce fluorescence detection to near shot-noise levels. High-photon flux excitation can overcome photon-limited shot...

Optical neurotechnologies use light to interface with neurons and can monitor and manipulate neural activity with high spatial-temporal precision over large cortical extents. While there has been significant progress in miniaturizing microscope for head-mounted configurations, these existing devices are still very bulky and could never be fully imp...

To understand the operation of the olfactory system, it is essential to know how information is encoded in the olfactory bulb. We applied Shannon information theoretic methods to address this, with signals from up to 57 simultaneously optically imaged from pre-synaptic inputs in glomeruli in the mouse dorsal and lateral olfactory bulb, in response...

Genetically encoded fluorescent voltage indicators are ideally suited to reveal the millisecond-scale interactions among and between targeted cell populations. However, current indicators lack the requisite sensitivity for in vivo multipopulation imaging. We describe next-generation green and red voltage sensors, Ace-mNeon2 and VARNAM2, and their r...

To survive in dynamic environments, animals make behavioral decisions based on innate and learnt information about the valences of sensory cues. In the brains of multiple species, innate and learnt sensory valence signals are initially encoded by distinct neural populations1–7 but then reconverge in downstream brain structures that guide behavioral...

Monitoring spiking activity across large neuronal populations at behaviorally relevant timescales is critical for understanding neural circuit function. Unlike calcium imaging, voltage imaging requires kilohertz sampling rates which reduces fluorescence detection to near shot noise levels. High-photon flux excitation can overcome photon-limited sho...

Genetically encoded fluorescent voltage indicators are ideally suited to reveal the millisecond-scale interactions among and between distinct, targeted cell populations. However, current indicator families lack the requisite sensitivity for in vivo multipopulation imaging. We describe high-performance green and red sensors, Ace-mNeon2 and VARNAM2,...

Pyrosomes are tunicates in the phylum Chordata, which also contains vertebrates. Their gigantic blooms play important ecological and biogeochemical roles in oceans. Pyrosoma, meaning “fire-body”, derives from their brilliant bioluminescence. The biochemistry of this light production is unknown, but has been hypothesized to be bacterial in origin. W...

Genetically encoded voltage indicators (GEVIs) allow for cell-specific optical recordings of membrane potential changes in defined cell populations. One tool that would further their use in the in vivo mammalian brain is transgenic reporter animals that facilitate precise and repeatable targeting with high expression levels. The present literature...

The latest generation of genetically encoded voltage indicators (GEVIs) is significantly advancing our ability to study electrical activity from large numbers of identified neurons. The further refinement of the technology will contribute to our understanding of behavior-evoked information perception, transfer and processing on a cellular level acr...

The mammalian olfactory bulb (OB) plays an essential role in odor processing during the perception of smell. Optical imaging of the OB has proven to be a key tool in elucidating the spatial odor mapping and temporal dynamics that underlie higher-order odor processing. Much is known about the activation of olfactory sensory neuron (OSN) glomerular r...

Schooling fishes, like flocking birds and swarming insects, display remarkable behavioral coordination. While over 25% of fish species exhibit schooling behavior, nighttime schooling has rarely been observed or reported. This is due to vision being the primary modality for schooling, which is corroborated by the fact that most fish schools disperse...

Although in recent years there has been an increased awareness of the widespread nature of biofluorescence in the marine environment, the diversity of the molecules responsible for this luminescent phenotype has been mostly limited to green fluorescent proteins (GFPs), GFP-like proteins, and fluorescent fatty acid-binding proteins (FABPs). In the p...

Genetically encoded optical indicators of neuronal activity enable unambiguous recordings of input-output activity patterns from identified cells in intact circuits. Among them, genetically encoded voltage indicators (GEVIs) offer additional advantages over calcium indicators as they are direct sensors of membrane potential and can adeptly report s...

Genetically encoded voltage indicators (GEVIs) are emerging optical tools for acquiring brain-wide cell-type-specific functional data at unparalleled temporal resolution. To broaden the application of GEVIs in high-speed multispectral imaging, we used a high-throughput strategy to develop voltage-activated red neuronal activity monitor (VARNAM), a...

Modern marine biologists seeking to study or interact with deep-sea organisms are confronted with few options beyond industrial robotic arms, claws, and suction samplers. This limits biological interactions to a subset of "rugged" and mostly immotile fauna. As the deep sea is one of the most biologically diverse and least studied ecosystems on the...

Bioluminescent copepods are often the most abundant marine zooplankton and play critical roles in oceanic food webs. Metridia copepods exhibit particularly bright bioluminescence, and the molecular basis of their light production has just recently begun to be explored. Here we add to this body of work by transcriptomically profiling Metridia lucens...

Fasta file with putative luciferase sequences

Fasta file of COI alignment for Metridia species Samples are identifiable by GenBank accession numbers.

Video of bioluminescent Metridia lucens Specimens first under normal light and then while excreting bioluminesce.

Tree file of COI phylogeny for Metridia species Samples are identifiable by GenBank accession numbers.

Fasta file with mature C-terminus “FYY” sequences

In order to understand how brain activity produces adaptive behavior we need large-scale, high-resolution recordings of neuronal activity. Fluorescent genetically encoded voltage indicators (GEVIs) offer the potential for these recordings to be performed chronically from targeted cells in a minimally invasive manner. As the number of GEVIs successf...

Genetically encoded calcium indicators (GECIs) produce unprecedentedly large signals that have enabled routine optical recording of single neuron activity in vivo in rodent brain. Genetically encoded voltage indicators (GEVIs) offer a more direct measure of neuronal electrical status, however the signal to noise characteristics and signal polarity...

Biofluorescence has recently been found to be widespread in marine fishes, including sharks. Catsharks, such as the Swell Shark (Cephaloscyllium ventriosum) from the eastern Pacific and the Chain Catshark (Scyliorhinus retifer) from the western Atlantic, are known to exhibit bright green fluorescence. We examined the spectral sensitivity and visual...

Hypothesis: Cerebrospinal fluid (CSF) can be identified by using an enzyme-linked immunosorbent assay (ELISA) for Dickkopf-related protein 3 (DKK3). Background: Cerebrospinal fluid leakage from the subarachnoid space is a potentially alarming condition that, left unrepaired, may result in increased risk of meningitis and encephalitis. Current bi...

Optical imaging of voltage indicators based on green fluorescent proteins (FPs) or archaerhodopsin has emerged as a powerful approach for detecting the activity of many individual neurons with high spatial and temporal resolution. Relative to green FP-based voltage indicators, a bright red-shifted FP-based voltage indicator has the intrinsic advant...

We report the identification and characterization of two new members of a family of bilirubin-inducible fluorescent proteins (FPs) from marine chlopsid eels and demonstrate a key region of the sequence that serves as an evolutionary switch from non-fluorescent to fluorescent fatty acid-binding proteins (FABPs). Using transcriptomic analysis of two...

The amphinomid polychaete Hermodice carunculata is a cosmopolitan and ecologically important omnivore in coral reef ecosystems, preying on a diverse suite of reef organisms and potentially acting as a vector for coral disease. While amphinomids are a key group for determining the root of the Annelida, their phylogenetic position has been difficult...

Background: The amphinomid polychaete Hermodice carunculata is a cosmopolitan and ecologically important omnivore in coral reef ecosystems, preying on a diverse suite of reef organisms and potentially acting as a vector for coral disease. While amphinomids are a key group for determining the root of the Annelida, their phylogenetic position has bee...

Significance A previously unidentified set of thermosensory neurons embedded in the olfactory organ of the Drosophila larva is shown to be required to drive the animal up temperature gradients toward preferred environments. Optogenetics and optical neurophysiology reveal efficient sensory encoding of both favorable (warming) and unfavorable (coolin...

ArcLight, a genetically encoded fluorescent protein voltage probe with a large ΔF/ΔV, is a fusion between the voltage sensing domain of the Ciona instestinalis voltage sensitive phosphatase and super ecliptic pHluorin carrying a single mutation (A227D in the fluorescent protein). Without this mutation the probe produces only a very small change in...

The present invention concerns novel isolated fluorescent proteins, variants thereof, and polynucleotides encoding the same. Methods for making and using the polypeptides and polynucleotides are also provided. For example, methods to detect protein-protein interactions, to develop novel fluorescent reagents, to monitor cellular events, as well as c...

The need for effective genetically-encoded voltage indicators is widely recognized. Several prototypes have been made to date by fusing one or more fluorescent proteins to a voltage sensitive membrane protein (1). However, one factor that has limited the utility of these indicators is poor voltage sensitivity, making it a challenge to express the i...

The discovery of fluorescent proteins has revolutionized experimental biology. Whereas the majority of fluorescent proteins have been identified from cnidarians, recently several fluorescent proteins have been isolated across the animal tree of life. Here we show that biofluorescence is not only phylogenetically widespread, but is also phenotypical...

We previously reported the discovery of a fluorescent protein voltage probe, ArcLight, and its derivatives that exhibit large changes in fluorescence intensity in response to changes of plasma membrane voltage. ArcLight allows the reliable detection of single action potentials and sub-threshold activities in individual neurons and dendrites. The re...

Genomic and transcriptomic sequence data are essential tools for tackling ecological problems. Using an approach that combines next-generation sequencing, de novo transcriptome assembly, gene annotation and synthetic gene construction, we identify and cluster the protein families from Favia corals from the northern Red Sea. We obtained 80 million 7...

Nervous systems process information by integrating the electrical activity of neurons in complex networks. This motivates the long-standing interest in using optical methods to simultaneously monitor the membrane potential of multiple genetically targeted neurons via expression of genetically encoded fluorescent voltage indicators (GEVIs) in intact...

In this paper we present a fixed window level crossing sampling analog to digital convertor for bio-potential recording sensors. This is the first proposed and fully implemented fixed window level crossing ADC without local DACs and clocks. The circuit is designed to reduce data size, power, and silicon area in future wireless neurophysiological se...

Arclight is a fusion protein of superEcliptic pHluorin-A227D fluorescent protein and the voltage sensing domain of the Ciona voltage sensitive phosphatase. It exhibits a fluorescence intensity decrease of 40% in response to a 100 mV-depolarization from −70 mV holding potential. Its large optical signal allows it to detect action potentials and sub-...

Optogenetic reporters of membrane potential allow for recording of genetically distinct populations of neurons, although their usefulness to date has been limited by poor in vivo expression, small signal sizes and slow kinetics. The recently developed fluorescent protein (FP) voltage probe ArcLight exhibits a change in fluorescence to a 100 mV depo...

In this paper we present a fully self-contained imaging instrument (30 mm overall length) that is capable of recording high speed and detect relatively small fluorescent signals (0.1% ΔF/F) from brain tissues potentially containing genetically-encoded sensors or dyes. This device potentially enables the study of neuronal activity in awake and mobil...

There is a pressing need in neuroscience for genetically-encoded, fluorescent voltage probes that can be targeted to specific neurons and circuits to allow study of neural activity using fluorescent imaging. We created 90 constructs in which the voltage sensing portion (S1-S4) of Ciona intestinalis voltage sensitive phosphatase (CiVSP) was fused to...

Monitoring neuronal electrical activity using fluorescent protein-based voltage sensors has been limited by small response magnitudes and slow kinetics of existing probes. Here we report the development of a fluorescent protein voltage sensor, named ArcLight, and derivative probes that exhibit large changes in fluorescence intensity in response to...

The movie illustrates the optical response of an action potential (AP) in a pyramidal neuron from a hippocampal cell culture expressing the ElectricPK probe. The value of each pixel is the spike-triggered average value (over 43 action potentials) aligned to the peak of the action potential in the somatic whole cell recording. First, a low pass temp...

A substantial increase in the speed of the optical response of genetically encoded fluorescent protein voltage sensors (FP voltage sensors) was achieved by using the voltage-sensing phosphatase genes of Nematostella vectensis and Danio rerio. A potential N. vectensis voltage-sensing phosphatase was identified in silico. The voltage-sensing domain (...

An updated image sensor to be used in a miniature microscope system for recording brain activity over a wide cortical area (4–9 mm2) is presented. The image sensor is a 100 × 100 pixel array fabricated in bulk CMOS process. The sensor also contains a 10-bit ADC and two input channels to measure other physiological signals concurrently with the opti...

We improved a genetically encoded voltage sensitive optical probe, a conjugate of the Ecliptic GFP and CiVSP, by modifying the fluorescent protein and its insertion site in the voltage sensitive phosphatase. The signal size is increased from −1% to −35% for 100 mV depolarization steps. The improved probes’ dynamics is fitted by the double exponenti...

We examined the effect of large, controlled whisker movements, delivered at a high speed, on the amplitude and spread of depolarization in the anesthetized mouse barrel cortex. The stimulus speed was varied between 1500 and 6000°/s and the extent of movement was varied between 4° and 16°. The rate of rise of the response was linearly related to the...

In this paper we present a rodent head-mountable, fully self contained imaging device that is capable of recording high speed fluorescent signals from genetically-encoded probes or voltage sensitive dyes. This device enables the study of neuronal activity in awake and mobile animals during more natural behaviors without the stress and suppression o...

FlaSh-YFP, a fluorescent protein (FP) voltage sensor that is a fusion of the Shaker potassium channel with yellow fluorescent protein (YFP), is primarily expressed in the endoplasmic reticulum (ER) of mammalian cells, possibly due to misfolded monomers. In an effort to improve plasma membrane expression, the FP was split into two non-fluorescent ha...

We report a head-mountable CMOS camera for recording rapid neuronal activity in freely moving rodents using fluorescent activity reporters. This small, lightweight camera is capable of detecting small changes in light intensity (0.2% ΔI/I) at 500fps. The camera has a resolution of 32×32, sensitivity of 0.62V/lxs, conversion gain of 0.52μV/e(-) and...

Organic voltage-sensitive dyes offer very high spatial and temporal resolution for imaging neuronal function. Further progress in imaging activity is expected from the emergent development of genetically encoded fluorescent sensors of membrane potential. These fluorescent protein (FP) voltage sensors overcome some drawbacks of organic voltage sensi...

Fluorescent proteins have become essential tools in molecular and biological applications. Here, we present a novel fluorescent protein isolated from warm water coral, Cyphastrea microphthalma. The protein, which we named vivid Verde fluorescent protein (VFP), matures readily at 37 °C and emits bright green light. Further characterizations revealed...

Imaging activity of neurons in intact brain tissue was conceived several decades ago, and, after many years of development, voltage-sensitive dyes now offer the highest spatial and temporal resolution for imaging neuronal functions in the living brain. Further progress in this field is expected from the emergent development of genetically encoded f...

Over the past decade, fluorescent proteins (FPs) have become ubiquitous tools in biological research. Yet, little is known about the natural function or evolution of this superfamily of proteins that originate from marine organisms. Using molecular phylogenetic analyses of 102 naturally occurring cyan fluorescent proteins, green fluorescent protein...