Research Departments & Organizations
Dr Pieribone is developing genetically encoded fluorescent probes of membrane electrical potential. These probes allow one to use optical instruments (microscopes) to monitor the electrical activity of neurons. Such an approach is less invasive, allows study of identified cell types over large regions of the cortical surface. The laboratory has also engineered miniature imaging systems that can be head mounted on mammels and allow mobile recording of neuronal activity. These types of studies will allow a better understanding of the neuronal networks that encode information in the central nervous system.
Specialized Terms: Neurophysiology; Neurotransmission; Voltage and calcium imaging; Sensory physiology; Drug development; Coral biology; Fluorescent proteins
Extensive Research Description
My published research has spanned 26 years (beginning with a Science paper published in 1986 while I was an undergraduate), produced >100 publications in peer-reviewed journals and I co-authored a well received (Nature, Technology or Mixed Signals Blog, CBE Life Science Education, Oceanography, Development, Yale Scientific and Nature Cell Biology), original peer-reviewed lay scientific book (“Aglow in the dark: the revolutionary science of biofluorescence” Harvard Press). My research has spanned a number of scientific areas: catecholamine anatomy and physiology, neuropeptide anatomy and physiology, adrenoceptor distribution in the central nervous system, serotonergic anatomy, the physiology and biochemistry of synaptic transmission, development of genetically-encoded voltage probes, identification of fluorescent proteins in marine animals, and information content in barrel cortex voltage-dye imaging. My work has received some 8,900 citations, an average of 58 per publication and 181 per year since 1986 (h-index of 47).
As an undergraduate and graduate student I published a series of papers on the afferent control of the locus coeruleus. These publications developed a novel and still held, concept that control of the brain’s main norepinephrine system largely resided in the hind brain. This concept realigned the fields perception of what factors controlled this important brain system. The locus coeruleus/norepinephrine system is involved in the actions of a number of widely used psychopharmacological interventions for depression, mania and schizophrenia.
As a Fogarty and National Science Foundation Fellow at the Nobel Institute of Neurophysiology in Stockholm Sweden, I published a series of studies on the anatomical distribution of the newly cloned adrenoceptors. These studies were the first to accurately identify which cells in the brain (and rest of the body) expressed which adrenoceptors. Previous receptor binding studies could not identify which cells or which cellular elements produced the labeled receptors and the specificity of the labeling was always questionable. These studies remain the most complete mapping of adrenoceptor subtypes in the brain (and other organs). While in Sweden I also examined the neurophysiologic response of monoaminergic neurons to the neuropeptide galanin and nitrous oxide. These represent one of only a handful of studies examining the neurophysiologic effects of galanin.
At The Rockefeller University I developed an in situ experimental preparation of an isolated large synapse for use in biochemical studies of neurotransmitter release. My research focused on the role of synapsin and actin in neurotransmission. We established that acute removal of the synaptic vesicle protein, synapsin from an adult synapses causes little change in low frequency neurotransmitter release but highly impairs high frequency neurotransmitter release. These studies combined with corroborative electron microscopic data, strongly suggested that the role of the most prevalent synaptic vesicle protein known is to create a reserve of synaptic vesicles that can be rapidly mobilized during bouts of high activity. The loss of this control was later shown to produce seizures in adult mice and in humans. At The Pierce Laboratory/Yale University I continued my work on synaptic transmission and moved to study the role of actin and actin- binding proteins on synaptic ultrastructure and physiology. In a series of papers we identified a clear cut role for actin, synapsins and talin in the recycling and clustering of synaptic vesicles.
In 2001 I began a research project to develop a genetically-encoded voltage probe based on green fluorescent protein. Being able to record electrical activity in virtually any specific cell type non-invasively will revolutionize neuroscience. In 2002 we published one of three original proof-of-concept genetically-encoded voltage probes to be developed by the scientific community. Our probe was the first to show rapid on and off kinetics. This publication led to an NIH R21 grants to explore other probes. At that time we also began to look for new fluorescent proteins that would preform better in voltage probes. We have begun a program to clone fluorescent proteins from coral. Our voltage probe became a strong proof-of- concept for the production of such probes. We subsequently received a multicenter (Yale, Riken (Japan), U. Montana, U. Penn. and U.C. Berkeley) NIH U24 grant that involved analyzing a large protein space for improved probes. This grant resulted the discovery and publication from my laboratory in September 2012, of a major advance in optical recording of brain activity (Jin, Lei et al. “Single Action Potentials and Subthreshold Electrical Events Imaged in Neurons with a Fluorescent Protein Voltage Probe.” Neuron 75.5 (2012): 779–785.) This was followed by the first ever fully optical neurophysiology study (Cao G, Platisa J, Pieribone VA, Raccuglia D, Kunst M, Nitabach MN. Genetically targeted optical electrophysiology in intact neural circuits. Cell 154: 904–913, 2013.). This work has received considerable attention in relevant media (BioTechniques, Yale Medicine, Yale Daily News, Science Daily, Scientific American, National Geographic, NIH News, Reddit).
My laboratory has also developed the first head-mountable, high speed, fluorescent microscope system for monitoring electrical activity using genetically-encoded sensors of voltage and calcium. This microscope is now being used in combination with the above mentioned probe to study wide-spread motor cortical activity in an awake behaving animal with high temporal resolution. We are developing a fully optical neural prosthesis capable of informing accurate positional information in realtime. These studies are aimed at eventual human implementation in paralyzed patients.
I have been intimately involved in the development and Phase II/III clinical testing of the neurosteroid ganaxolone and allopregnanolone (I am the author of clinical grant applications, a peer-review paper and clinical protocols). I have co-developed the three clinical trials for these compounds in epilepsy ,post traumatic stress disorder and traumatic brain injury. I consider the development of neuropharmaceuticals a natural extension of my laboratory work at Yale University and the ultimate and most significant application of basic science to the betterment of human health.
I am currently the President and co-founder of Affinimark Technologies, Inc. which is developing a rapid point-of-care test for the presence of cerebrospinal fluid in bodily fluids leaked from the head. No such test exists and would improve patient care following traumatic head injury and sinus/ear surgery.
My scientific achievements extend well beyond my laboratory science. I am a Research Scientist at both the Mystic Aquarium and Center for Exploration in Mystic Connecticut and the American Museum of Natural History in New York City. I have co-curated a permanent exhibit at the Mystic Aquarium (http://www.mysticaquarium.org/animals-and-exhibits/exhibits/indoor/259-fluorescent-corals) and the most popular temporary exhibit to date at the American Museum of Natural History (- http://ez-www.amnh.org/creatures-of-light/ , National Science Foundation story ). For the American Museum of Natural History exhibit we received an National Science Foundation education grant. I have led several research expeditions to study and collect biofluorescent animals from coral reefs in Belize, British Virgin Islands, Malaysia, Australia, Israel, and the Solomon Islands. These have been funded by the National Institutes of Health, National Geographic Society, National Science Foundation, EarthWatch, and the American Museum of Natural History). Accounts of these trips have been documented in televised documentaries, radio interviews (Science and Society, National Public Radio, Studio 360, National Public Radio) and in web (National Geographic, Good News, Big Think, World Science Festival, Live Science) and print articles including The New York Times ( http://scientistatwork.blogs.nytimes.com/author/vincent-pieribone/). In 2010, we (along with my collaborator David Gruber) received an NSF Major Research Instrumentation grant to develop a remotely operated vehicle (ROV) to study deep sea fluorescent and bioluminescent animals.
The covert world of fish biofluorescence: a phylogenetically widespread and phenotypically variable phenomenon.
Sparks JS, Schelly RC, Smith WL, Davis MP, Tchernov D, Pieribone VA, Gruber DF. The covert world of fish biofluorescence: a phylogenetically widespread and phenotypically variable phenomenon. PloS One 2014, 9:e83259. 2014
Genetically targeted optical electrophysiology in intact neural circuits.
Cao G, Platisa J, Pieribone VA, Raccuglia D, Kunst M, Nitabach MN. Genetically targeted optical electrophysiology in intact neural circuits. Cell 2013, 154:904-13. 2013
Fluorescent protein voltage probes derived from ArcLight that respond to membrane voltage changes with fast kinetics.
Han Z, Jin L, Platisa J, Cohen LB, Baker BJ, Pieribone VA. Fluorescent protein voltage probes derived from ArcLight that respond to membrane voltage changes with fast kinetics. PloS One 2013, 8:e81295. 2013
A fluorescent, genetically-encoded voltage probe capable of resolving action potentials.
Barnett L, Platisa J, Popovic M, Pieribone VA, Hughes T. A fluorescent, genetically-encoded voltage probe capable of resolving action potentials. PloS One 2012, 7:e43454. 2012
Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe.
Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA. Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe. Neuron 2012, 75:779-85. 2012
Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics.
Baker BJ, Jin L, Han Z, Cohen LB, Popovic M, Platisa J, Pieribone V. Genetically encoded fluorescent voltage sensors using the voltage-sensing domain of Nematostella and Danio phosphatases exhibit fast kinetics. Journal Of Neuroscience Methods 2012, 208:190-6. 2012
Random insertion of split-cans of the fluorescent protein venus into Shaker channels yields voltage sensitive probes with improved membrane localization in mammalian cells.
Jin L, Baker B, Mealer R, Cohen L, Pieribone V, Pralle A, Hughes T. Random insertion of split-cans of the fluorescent protein venus into Shaker channels yields voltage sensitive probes with improved membrane localization in mammalian cells. Journal Of Neuroscience Methods 2011, 199:1-9. 2011
Head-mountable high speed camera for optical neural recording.
Park JH, Platisa J, Verhagen JV, Gautam SH, Osman A, Kim D, Pieribone VA, Culurciello E. Head-mountable high speed camera for optical neural recording. Journal Of Neuroscience Methods 2011, 201:290-5. 2011
Effect of high velocity, large amplitude stimuli on the spread of depolarization in S1 "barrel" cortex.
Davis DJ, Sachdev R, Pieribone VA. Effect of high velocity, large amplitude stimuli on the spread of depolarization in S1 "barrel" cortex. Somatosensory & Motor Research 2011, 28:73-85. 2011
A new bright green-emitting fluorescent protein--engineered monomeric and dimeric forms.
Ilagan RP, Rhoades E, Gruber DF, Kao HT, Pieribone VA, Regan L. A new bright green-emitting fluorescent protein--engineered monomeric and dimeric forms. The FEBS Journal 2010, 277:1967-78. 2010
Novel internal regions of fluorescent proteins undergo divergent evolutionary patterns.
Gruber DF, DeSalle R, Lienau EK, Tchernov D, Pieribone VA, Kao HT. Novel internal regions of fluorescent proteins undergo divergent evolutionary patterns. Molecular Biology And Evolution 2009, 26:2841-8. 2009
Genetically encoded fluorescent sensors of membrane potential.
Baker BJ, Mutoh H, Dimitrov D, Akemann W, Perron A, Iwamoto Y, Jin L, Cohen LB, Isacoff EY, Pieribone VA, Hughes T, Knöpfel T. Genetically encoded fluorescent sensors of membrane potential. Brain Cell Biology 2008, 36:53-67. 2008
Patterns of fluorescent protein expression in Scleractinian corals.
Gruber DF, Kao HT, Janoschka S, Tsai J, Pieribone VA. Patterns of fluorescent protein expression in Scleractinian corals. The Biological Bulletin 2008, 215:143-54. 2008
Early involvement of synapsin III in neural progenitor cell development in the adult hippocampus.
Kao HT, Li P, Chao HM, Janoschka S, Pham K, Feng J, Mcewen BS, Greengard P, Pieribone VA, Porton B. Early involvement of synapsin III in neural progenitor cell development in the adult hippocampus. The Journal Of Comparative Neurology 2008, 507:1860-70. 2008
Dynamic regulation of fluorescent proteins from a single species of coral.
Kao HT, Sturgis S, DeSalle R, Tsai J, Davis D, Gruber DF, Pieribone VA. Dynamic regulation of fluorescent proteins from a single species of coral. Marine Biotechnology (New York, N.Y.) 2007, 9:733-46. 2007
In vivo simultaneous tracing and Ca(2+) imaging of local neuronal circuits.
Nagayama S, Zeng S, Xiong W, Fletcher ML, Masurkar AV, Davis DJ, Pieribone VA, Chen WR. In vivo simultaneous tracing and Ca(2+) imaging of local neuronal circuits. Neuron 2007, 53:789-803. 2007
A genetically targetable fluorescent probe of channel gating with rapid kinetics.
Ataka K, Pieribone VA. A genetically targetable fluorescent probe of channel gating with rapid kinetics. Biophysical Journal 2002, 82:509-16. 2002
Midbrain serotonergic neurons are central pH chemoreceptors.
Severson CA, Wang W, Pieribone VA, Dohle CI, Richerson GB. Midbrain serotonergic neurons are central pH chemoreceptors. Nature Neuroscience 2003, 6:1139-40. 2003
Genetically encoded fluorescent voltage indicators: are we there yet?
Platisa J, Pieribone VA. Genetically encoded fluorescent voltage indicators: are we there yet? Current Opinion In Neurobiology 2018, 50:146-153. 2018
Directed Evolution of Key Residues in Fluorescent Protein Inverses the Polarity of Voltage Sensitivity in the Genetically Encoded Indicator ArcLight.
Platisa J, Vasan G, Yang A, Pieribone VA. Directed Evolution of Key Residues in Fluorescent Protein Inverses the Polarity of Voltage Sensitivity in the Genetically Encoded Indicator ArcLight. ACS Chemical Neuroscience 2017, 8:513-523. 2017
Biofluorescence in Catsharks (Scyliorhinidae): Fundamental Description and Relevance for Elasmobranch Visual Ecology.
Gruber DF, Loew ER, Deheyn DD, Akkaynak D, Gaffney JP, Smith WL, Davis MP, Stern JH, Pieribone VA, Sparks JS. Biofluorescence in Catsharks (Scyliorhinidae): Fundamental Description and Relevance for Elasmobranch Visual Ecology. Scientific Reports 2016, 6:24751. 2016