2018 — 2019 |
Feller, Marla [⬀] Landry, Markita |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Novel Optical Probe For Dopamine Release in Neural Circuits @ University of California Berkeley
A major focus of modern neuroscience is understanding the role of neuromodulators in neural circuit function and development, in behavior, and their dysfunction in neurological disease. Neural circuit research has been revolutionized by powerful new optical techniques. Here we propose to use new near-infrared optical nanosensor technology to image dopamine. As a proof of principle, we will use this sensor to monitor dopamine release under physiological conditions in both the developing and adult retinas. The ability to combine neuromodulator imaging with other physiological measures in a well-defined neural circuit will greatly advance our understanding of neuromodulation, allowing us to elucidate its many impacts on normal and pathological circuits.
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2019 — 2020 |
Landry, Markita |
R35Activity Code Description: To provide long term support to an experienced investigator with an outstanding record of research productivity. This support is intended to encourage investigators to embark on long-term projects of unusual potential. |
Single-Molecule Imaging of Biological Trauma: Cytokine-Based Intracellular Communication @ University of California Berkeley
PROJECT ABSTRACT Research Abstract: The detection of unlabeled cytokines in real-time and from single cells could provide a robust platform for understanding the ?molecular language of biological trauma and disease?. However, tools to visualize cytokines at the cellular level, particularly in their secreted form, are lacking. We have developed a generic nanomaterial-based near-infrared fluorescent sensor and accompanying microscopy platform which produces a unique intensity and wavelength shift in the presence of a specific target molecule (Zhang*, Landry* et al. Nature Nanotechnology 2013; Landry et al. Sensors 2015; Landry et al. Nature Nanotechnology 2017). In this 5-year proposal, I (i) will develop synthetic sensors for VEGF, IL-6, and IL-8 cytokines, (ii) validate their use to monitor constitutive cytokine secretion from macrophage and epithelial cells, and (iii) directly visualize the spatio-temporal profiles of intercellular cytokine-based synergies. Direct cellular measurement of secreted cytokines will inform how cytokine secretion profiles from single or few individual cells are stimulated by chemokines and cytokines, which forms the basis of the cytokine secretion profiles currently used in biomarker- based diagnostics. The research we propose herein has ? to the best of our knowledge ? only been explored theoretically (Thurley et al. POLS Comp. Bio. 2015). Landry Laboratory Research Program: I am a single-molecule biophysicist by training, having developed several instruments capable of detecting piconewton-scale forces (Landry et al. Biophys. J. 2009), and nanometer-scale fluorescence localization (Landry et al. Nucl. Ac. Res. 2012) for my doctoral work. In transitioning to my postdoctoral position, my goal was to leverage my expertise in single-molecule spectroscopy and molecular biophysics to design purely synthetic molecular recognition tools. My scientific training in as a postdoctoral fellow in Chemical Engineering at MIT focused on merging these two previously disparate areas of science: optical microscopy and nanosensor development, yielding a platform for the optical detection of any generic molecular analyte. I began my faculty appointment at UC Berkeley in June 2016, with a research portfolio motivated by translating the technical strengths of my lab in microscopy (O?Donnel et al. Adv. Funct. Mater 2017), sensor development (Beyene et al. ACS Chem Neruo 2017 & Luo et al. ACS Sensors 2017), and molecular recognition (Li et al. RSC Chemical Science 2017) to addressing the need to develop methods to detect cytokine efflux from immune cells. In the first two years of my research plan, my group will synthesize and characterize nanomaterial-based sensors for cytokines in vitro. The remaining three years of the R35 award will implement the use of cytokine sensors to measure constitutive (year 3), induced (year 4), and intracellular (year 5) cytokine signaling from cultured cell samples. My long-term research goals focus on the application of nanosensors for cytokines, chemokines, and other important biomarkers in environments such as multicellular tumor spheroids and live tumor biopsies, in which biomarker detection has traditionally been difficult. Landry ABSTRACT AB-1
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2021 |
Landry, Markita |
R03Activity Code Description: To provide research support specifically limited in time and amount for studies in categorical program areas. Small grants provide flexibility for initiating studies which are generally for preliminary short-term projects and are non-renewable. |
Imaging Presynaptic Correlates of Cocaine Locomotor Sensitization With Near Infrared Catecholamine Osensors @ University of California Berkeley
PI: Markita Landry (Assistant Professor, UC Berkeley Chemical and Biomolecular Engineering) Project Abstract Imaging presynaptic correlates of cocaine locomotor sensitization with near infrared catecholamine nanosensors Cocaine locomotor sensitization is thought to be, in part, driven by changes in dopamine signaling within the nucleus accumbens (NAc). One potentially early contributing mechanism is internalization and degradation of D2 autoreceptors within dopamine neurons (Madhavan et al. 2013) and their long range processes. We recently developed a technique for imaging dopamine neurotransmission in the striatum using synthetic near infrared catecholamine nanosensors (nIRCats) (Beyene et al. 2019). nIRCats are compatible with a wide variety of dopamine receptor drugs, and can resolve individual dopamine release ?hotspots? ~2 µm in size. We aim to determine how cocaine locomotor sensitization affects D2 autoreceptor regulation of dopamine release at individual nIRCat hotspots. Adult male & female C57/Bl6 mice will undergo a five day locomotor sensitization protocol (control: saline; cocaine: 15 mg/kg). Twenty-four hours after the final locomotor testing session, acute brain slices will be prepared from saline or cocaine-treated mice, and will be labeled with nIRCat. Dopamine release will be electrically evoked within the NAc or dorsal striatum to image dopamine release spatiotemporal kinetics. Via bath application of 1 µM quinpirole and repeating the stimulation protocol, we will assess the change in evoked nIRCat ?F/F to assay presynaptic D2 autoreceptor function. By thus probing dopamine autoreceptor response to quinpirole, we will compare drug effects across treatment conditions and striatal subregions to answer if and how repeated cocaine administration influences the regulation of presynaptic dopamine release by D2 autoreceptors. We predict that quinpirole will show a reduced ability to suppress dopamine release in cocaine-sensitized animals relative to the saline control, and will explore differences in sensitization across individual dopamine release sites in striatum and NAc of sensitized animals. This work will establish near infrared imaging, nIRCat nanosensors, and pharmacology as valuable tools to measure dopamine transients at the level of individual synapses and to analyze the relationship between their modulation by D2 receptor drugs and locomotor sensitization. Landry ABSTRACT AB-1
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2021 — 2024 |
Landry, Markita Vukovic, Lela [⬀] |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Learning and Characterizing Dna Sequences in Dna-Nanotube Hybrid Structures For High-Affinity Binding and Recognition of Molecular Targets @ University of Texas At El Paso
Nanomaterials with unique and powerful optical properties have emerged as promising tools for imaging and sensing small biological molecules present in cells, tissues, and live organisms. Imaging these important molecules is hampered by the lack of sensing tools, limiting our ability to understand how certain biological molecules allow the brain to communicate with the body. This project will develop new experimental and theoretical tools to detect the presence of brain-relevant molecules, with a focus on two molecules called serotonin and oxytocin, which are involved in human mood, emotions, social behavior, and their dysregulation in disorders like depression and autism. The new tools are based on DNA molecules, long molecules composed of repeating subunits, wrapped around long and rigid carbon nanotubes, which will detect serotonin and oxytocin in the sample by emitting light. The key objective is to discover special DNA molecules that will bind strongly to serotonin and oxytocin, bring them close to the nanotube surface, and change the optical signal that the nanotube emits. This objective will be addressed by combining the information obtained in experimental screening of DNA-nanotube systems with the artificial intelligence computer methods and computer simulations. The project will lead to new methods for discovery of useful DNA molecules, which could be applied towards developing tools to detect other biological molecules of interest that currently remain ‘invisible’. The efforts in this project will enrich the training and research experiences of underrepresented students at the University of Texas at El Paso and the University of California, Berkeley and provide the basis for demonstrations on science behind serotonin and oxytocin to middle school student groups.
Sensing and imaging of small molecular analytes within complex biological samples is of high interest but remains a challenge due to the lack of suitable imaging tools. This project will establish methods to systematically develop conjugates of single-walled carbon nanotubes and single stranded DNA for optical sensing of selected analytes. The methods will be established for two analytes, serotonin, and oxytocin brain neuromodulators. To develop new DNA-nanotube optical sensors, unique DNA sequences, which simultaneously bind with high affinity to the analytes on nanotube surfaces and lead to strong optical response of the nanotubes, need to be identified. Yet, discovering these useful DNA sequences has been a challenge, with most research efforts relying on screening-based serendipity. In this project, the research team will combine the high throughput datasets of experimental DNA sequence screening methods, lower throughput spectroscopic measurements, and the artificial intelligence-based methodology to learn and predict DNA sequences that bind with high affinity to analytes on nanotube surfaces and induce high nanotube optical activity. First, the team will develop and validate new methods to: 1) determine short DNA sequences that bind with high affinity to serotonin and oxytocin molecules on nanotube surfaces; and 2) learn and predict DNA sequences that provide high optical response to DNA-nanotube conjugates in the presence of the analytes. Then, the molecular basis of analyte recognition by DNA sequences on nanotube surfaces will be explored with simulations. This project should have a significant impact on many individuals from underrepresented groups at the University of Texas at El Paso and the University of California, Berkeley through several activities that will be organized. The computational chemistry modules will be introduced into physical chemistry laboratory classes, a student-coordinated research event will be organized, and demonstrations on the science behind serotonin and oxytocin will be developed for middle school student groups. Undergraduate students from underrepresented groups will be recruited and trained in the above research.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
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