Regulation Trigeminal Neuron Differentiation&Projection
Qiufu Ma
Dana-farber Cancer Institute
44 Binney St
boston, Ma 02115
Grant 1R01DE013843-01A1 from National Institute Of Dental & Craniofacial Research IRG: ZRG1
Abstract: The trigeminal ganglia are responsible for sensory processing in the face, oral and nasal cavities. Injuries and malformations of this nerve are clinically and cosmetically devastating to humans. Two different tissues contribute to the formation of the trigeminal ganglia the ectodermal placode and the cranial neural crest. Most mechanoceptive neurons (trkC+ and trkB+) are derived from the placode, whereas the neural crest gives rise to all of the nociceptive and thermoceptive neurons (trkA+). The broad objectives of this research are to understand how these distinct classes of sensory neurons develop and how they innervate to the locales within the face. In preliminary studies, my colleagues and I have found that a single neural determination gene, Neurogenin1 (ngnl), is required to direct formation of the entire trigeminal ganglion. Research described in this proposal is primarily based on this finding. We will address three fundamental questions 1) how is the neural crest-derived neurogenesis regulated by extrinsic signals from the pioneer placodal neurons? 2) How are axons of crest-derived neurons projected to precise locales within the face? 3) What are the intrinsic molecular mechanisms by which sensory sublineages are specified? For the first question, we will determine if the potential of the trigeminal neural crest cells to form the trkB+ and trkC+ neurons is restricted by pioneer placodal neurons. We will also determine if specification of other neuronal features is dependent on this cell-to-cell interaction as well. To do this, we will create trigeminal crest ganglia that completely lack the pioneer placodal neurons, and then to examine how the remained crest-derived neurogenesis will be affected. For the second question, a testable hypothesis is suggested by the classic experiments of Victor Hamburger. Pioneer placode-derived neurons may play an essential role for axon pathfinding of the crest-derived neurons. This hypothesis will be tested by examining axonal projections of crest-derived neurons that develop in mice without placodal neurons. For the third question, we will use "Gene Family Differential Screening," a procedure developed by the investigator, to isolate regulatory molecules that are expressed in a subset of sensory neurons. These sublineage-specific genes will be candidate molecules that control cell subtype specification
Keywords: cell differentiation, cell growth regulation, neurogenesis, neuron, trigeminal nerve biological signal transduction, cell cell interaction, ganglion, gene expression, genetic recombination, genetic screening, molecular dynamics, neural crest, neuronal guidance, phenotype, transcription factor embryonic stem cell, gene targeting, immunocytochemistry, in situ hybridization, laboratory mouse, tissue /cell culture, transgenic animal
Project start date: 2001-03-01
Project end date: 2006-02-28
1R01DE013843-01A1 (2001): $260583
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Qiufu Ma
Regulation Trigeminal Neuron Differentiation&Projection
Qiufu Ma
Dana-farber Cancer Institute 44 Binney St Boston, Ma 02115
Grant 5R01DE013843-05 from National Institute Of Dental And Craniofacial Research IRG: ZRG1
Abstract: The trigeminal ganglia are responsible for sensory processing in the face, oral and nasal cavities. Injuries and malformations of this nerve are clinically and cosmetically devastating to humans. Two different tissues contribute to the formation of the trigeminal ganglia the ectodermal placode and the cranial neural crest. Most mechanoceptive neurons (trkC+ and trkB+) are derived from the placode, whereas the neural crest gives rise to all of the nociceptive and thermoceptive neurons (trkA+). The broad objectives of this research are to understand how these distinct classes of sensory neurons develop and how they innervate to the locales within the face. In preliminary studies, my colleagues and I have found that a single neural determination gene, Neurogenin1 (ngnl), is required to direct formation of the entire trigeminal ganglion. Research described in this proposal is primarily based on this finding. We will address three fundamental questions 1) how is the neural crest-derived neurogenesis regulated by extrinsic signals from the pioneer placodal neurons? 2) How are axons of crest-derived neurons projected to precise locales within the face? 3) What are the intrinsic molecular mechanisms by which sensory sublineages are specified? For the first question, we will determine if the potential of the trigeminal neural crest cells to form the trkB+ and trkC+ neurons is restricted by pioneer placodal neurons. We will also determine if specification of other neuronal features is dependent on this cell-to-cell interaction as well. To do this, we will create trigeminal crest ganglia that completely lack the pioneer placodal neurons, and then to examine how the remained crest-derived neurogenesis will be affected. For the second question, a testable hypothesis is suggested by the classic experiments of Victor Hamburger. Pioneer placode-derived neurons may play an essential role for axon pathfinding of the crest-derived neurons. This hypothesis will be tested by examining axonal projections of crest-derived neurons that develop in mice without placodal neurons. For the third question, we will use "Gene Family Differential Screening," a procedure developed by the investigator, to isolate regulatory molecules that are expressed in a subset of sensory neurons. These sublineage-specific genes will be candidate molecules that control cell subtype specification.
Keywords: cell differentiation, cell growth regulation, neurogenesis, neuron, trigeminal nerve, biological signal transduction, cell cell interaction, ganglion, gene expression, genetic recombination, genetic screening, molecular dynamics, neural crest, neuronal guidance, phenotype, transcription factor, embryonic stem cell, gene targeting, genetically modified animal, immunocytochemistry, in situ hybridization, laboratory mouse, tissue /cell culture
Project start date: 2001-03-01
Project end date: 2007-02-28
5R01DE013843-05 (2005): $257145
5R01DE013843-04 (2004): $257761
5R01DE013843-03 (2003): $258359
Regulation Of Glutamate And GABA Neuron Development
Qiufu Ma
Dana-farber Cancer Institute 44 Binney St Boston, Ma 02115
Grant 5R01NS047710-04 from National Institute Of Neurological Disorders And Stroke IRG: MDCN
Abstract: The mammalian nervous system is composed of thousands of distinct neuronal cell types. However, all of them are either excitatory or inhibitory. The principal excitatory and inhibitory neurotransmitters are the amino acids glutamate and GABA (gamma-aminobutyrate), respectively. During development, these two transmitters are specified in a mutually exclusive manner. The broad goal of this proposal is to understand the molecular mechanism that underlies this very important fate choice decision. In preliminary studies, we have focused upon an anatomically well-defined region of the developing nervous system - the dorsal horn of the spinal cord. We have found that the Tlx-class transcription factors have a dual function in cell fate choice promoting glutamate and suppressing GABA neuron development. Thus, in Tlx-null mice, glutamatergic sensory cells in the dorsal horn are transformed into GABAergic neurons. Our study plan builds upon this preliminary data. We have three specific aims. Aim 1 is to define the anatomical range of Tlx function in the glutamatergic versus GABAergic fate choices. Is this Tlx function confined to the dorsal horn of the spinal cord or does it extend to other Tlx-positive regions of the central nervous system - specifically, the sensory nuclei in the hindbrain? Addressing this question will determine whether binary specification of glutamate and GABA is a common theme in the nervous system and may also provide insight into why Tlx-null mice suffer a breathing problem that resembles human congenital hypoventilation syndrome. Aim 2 is to define the roles of Tlx proteins in the fate choice process. Our preliminary studies show that Tlx proteins are necessary for glutamate neuron development. Here we will use genetic gain-of-function to determine whether Tlx proteins are sufficient to specify glutamate transmitter phenotype in various brain areas. In addition, we will determine whether Tlx proteins directly or indirectly promote glutamatergic neuron differentiation. Aim 3 is to define the structural basis of Tlx proteins in suppression of GABA neuron differentiation. Within the human CNS, a disruption of the balance between excitation and inhibition underlies neurological disorders, such as epilepsy, schizophrenia, and pain disorders. Accordingly the studies described here will have practical overtones for the management of these diseases.
Keywords: gamma aminobutyrate, glutamate, neurogenesis, neuroregulation, protein structure function, transcription factor, DNA binding protein, cell differentiation, dorsal horn, neurogenetics, rhombencephalon, genetically modified animal, laboratory mouse, polymerase chain reaction
Project start date: 2004-01-01
Project end date: 2008-12-31
5R01NS047710-04 (2007): $317086
5R01NS047710-03 (2006): $327166
5R01NS047710-02 (2005): $335648
1R01NS047710-01 (2004): $336237
RUNXL TARGETS AND MODULATORS IN MANIFESTATION OF CHRONIC PAIN
Qiufu Ma
Dana-farber Cancer Institute, 44 Binney St, Boston, Ma 02115
Abstract: The long-term goal of Project 2 is to develop new targets for the treatment of chronic pain. Specifically, our research has been focusing on those core transcriptional programs that control nociceptor phenotypes and pain behaviors. In the previous funding cycle, we have compiled a genome-scale analysis of the expression of transcription factors (TFs) in the developing nervous system. From this screen, we identified a small number of TFs expressed in the pain circuitry. Subsequent genetic studies demonstrated that the runt class transcripfion factor Runxl is a key regulator of nociceptor development, and mice lacking Runxl exhibit a marked deficit in inflammatory pain and neuropathic pain. The experimental plan of this project is built on these preliminary studies, and we have three specific aims. Aim 1 is to determine the roles of Runxl in controlling two types of cancer pain pain induced by tumor growth or by chemotherapy. This aim is built on the facts that cancer pain is composed of both inflammatory and neuropathic pain components, and Runxl is required for these two types of chronic pain. Aim 2 is to determine Runxl targets that serve as candidates critical for neuropathic pain. This aim is built on the observation that Runxl activity at embryonic stages, rather than at postnatal stages, is required for neuropathic pain, implying that early Runxl targets are later required for the development of this type of chronic pain. Aim 3 is to determine signaling pathways that modulate Runxl expression in adult nociceptors. This aim is built on the finding that persistent Runxl activity is required for inflammatory pain. Accordingly, compounds capable of extinguishing Runxl expression may serve as new targets for inflammatory pain treatment. The studies of these aims will be enabled by the availability of various Runxl mutant mice and from the "Druggable Mechanisms Core" (the DMC), including high throughput single molecule DNA sequencing and bioinformatics analyses
Keywords: 1, 2-diaminocyclohexane platinum oxalate; 1, 2-diamminocyclohexane(trans-1)oxolatoplatinum(II); 1-OHP; 21+ years old; Adult; Bio-Informatics; Bioinformatics; Cancer Pain Management; Carcinoma, Epidermoid; Carcinoma, Planocellular; Carcinoma, Squamous; Cell Communication and Signaling; Cell Signaling; DNA Sequence; Data; Depression; Development; Diaminocyclohexane Oxalatoplatinum; Disease Outcome; Embryo; Embryonic; Exhibits; Funding; Genetic; Genome; Goals; Human, Adult; Impairment; Intracellular Communication and Signaling; Knock-out; Knockout; L-OHP cpd; Label; Lectin; Mammals, Mice; Medical; Mental Depression; Methods and Techniques; Methods, Other; Mice; Mice, Mutant Strains; Molecular; Murine; Mus; Mutant Strains Mice; NRVS-SYS; Nerve Cells; Nerve Unit; Nervous System; Nervous system structure; Neural Cell; Neurocyte; Neurologic Body System; Neurologic Organ System; Neurons; Nociceptors; Oxalatoplatin; Oxalatoplatinum; Pain; Pain Control; Pain Therapy; Pain management; Painful; Phenotype; Programs (PT); Programs [Publication Type]; Reagent; Research; Role; Sciatic Nerve; Series; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Squamous Cell Epithelioma; Squamous cell carcinoma; Staging; Structure of sciatic nerve; Techniques; Testing; Time; Validation; Work; [(1R, -2R)-1, 2-cyclohexanediamine-N, N`][oxalato (2--)-O, O`]platinum; adult animal; adult human (21+); base; biological signal transduction; cancer pain; chemotherapy; chronic pain; chronic painful condition; drug development; experiment; experimental research; experimental study; human disease; inflammatory neuropathic pain; inflammatory pain; mature animal; mouse development; mouse mutant; mutant; neuronal; neuropathic pain; oxalato (1R, 2R-cyclohexanediamine)platinum(II); oxalato (trans-l-1, 2-diaminocyclohexane)platinum(II); oxalato-(1, 2-cyclohexanediamine)platinum II; oxaliplatin; oxaliplatine; pain behavior; painful neuropathy; platinum(II)-1, 2-cyclohexanediamine oxalate; postnatal; programs; research study; sciatic nerve; single molecule; social role; trans-l DACH oxalatoplatinum; trans-l diaminocyclohexane oxalatoplatinum; transcription factor; tumor growth
Relevance: Pain management remains a major nnedical problem in a variety of human diseases. Chronic pain, moreover, is associated with worse disease outcome and depression. In the fullness of time, the work may allow us to identify new targets for chronic pain treatment
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
5P01NS047572-07_5831 (2010): $261555
2P01NS047572-06_5831 (2009): $256642
MOLECULAR CONTROL OF SPINAL RELAY SENSORY NEURON PHENOYPES AND PAIN BEHAVIORS
Qiufu Ma, Associate Professor
Dana-farber Cancer Institute, 44 Binney St, Boston, Ma 02115
Grant 5R01NS047710-07 from National Institute Of Neurological Disorders And Stroke
Abstract: The long-range goal of this competitive renewal of a previous R01 is to investigate genetic programs that control pain relay sensory neuron phenotypes in the dorsal spinal cord. During the previous funding interval, my colleagues and I have compiled a genome-scale expression map of transcription factors in the mouse nervous system. Subsequent genetic studies demonstrate that Tlx3, a homeobox class transcription factor, is a pivotal regulator of spinal relay sensory neurons, including specification of both glutamate and peptide neurotransmitters. Furthermore, persistent Tlx3 expression in adult animals is confined to superficial laminae, where putative pain relay neurons are located. The research described here builds upon this preliminary work. The goal of our research over the next five years is to illustrate the roles of Tlx3 in regulating spinal relay nociceptor phenotypes and pain behaviors and to gain insights into the molecular and cellular basis underlying pain perception. We have four specific Aims. Aim 1 is to determine the roles of Tlx3 in controlling the development of ascending projection neurons that are critical for pain perception. Aim 2 is to determine how dynamic Tlx3 expression controls lamina organization of the dorsal spinal cord. Aim 3 is determine the roles of Tlx3 in maintaining dorsal horn excitatory neuron phenotypes, thereby determining if Tlx3-mediated core transcription program is a potential target for pain treatment. Aim 4 is to determine the roles of Tlx3-dependent differentiation programs in controlling pain behaviors. Each of these aims is built upon a set of preliminary data that lead to a testable hypothesis. A panel of genetic tools that we have already developed will test the predictions of these hypotheses. Pain management remains a major medical problem in a variety of human diseases. Chronic pain, moreover, is associated with worse disease outcome and depression. In the fullness of time, the work may allow us to determine whether the Tlx3-mediated core transcriptional program is a valid and novel therapeutic target for pain management
Keywords: Affect; Afferent Neurons; Alleles; Allelomorphs; Behavior Control; Behavioral; Behavioral Manipulation; Brain; Data; Defect; Development; Disease Outcome; Dorsal; Dorsal Horn Cells; Encephalon; Encephalons; Exhibits; Funding; Gene Expression; Gene Transcription; Genetic; Genetic Algorithm; Genetic Programming; Genetic Transcription; Genome; Glutamates; Goals; Homeo Boxes; Homeobox; Knock-out; Knockout; Knockout Mice; L-Glutamate; Label; Lead; Locomotion; Mammals, Mice; Maps; Mediating; Medical; Medulla Spinalis; Methods and Techniques; Methods, Other; Mice; Mice, Knock-out; Mice, Knockout; Molecular; Murine; Mus; NRVS-SYS; Nerve Cells; Nerve Impulse Transmission; Nerve Transmission; Nerve Transmitter Substances; Nerve Unit; Nervous System; Nervous System, Brain; Nervous system structure; Neural Cell; Neurocyte; Neurologic Body System; Neurologic Organ System; Neuronal Transmission; Neurons; Neurons, Afferent; Neurons, Dorsal Horn; Neurons, Posterior Horn; Neurons, Sensory; Neurotransmitters; Nociception; Nociceptors; Null Mouse; Pain; Pain Control; Pain Therapy; Pain management; Painful; Pattern; Pb element; Peptides; Perception; Phenotype; Play; Population; Posterior Horn Cells; Prevention; Process; Programs (PT); Programs [Publication Type]; RNA Expression; Research; Role; Sensory Cell Afferent Neuron; Spinal; Spinal Cord; Staging; Techniques; Testing; Thalamic Nuclei; Time; Transcription; Transcription, Genetic; Work; adult animal; base; behavioral control; chronic pain; chronic painful condition; depression; dorsal horn; excitatory neuron; heavy metal Pb; heavy metal lead; human disease; innervation; insight; mature animal; molecular phenotype; nerve supply; neural circuit; neural circuitry; neuronal; neurotransmission; new therapeutic target; nociceptive; novel; pain behavior; programs; public health relevance; social role; tool; transcription factor
Relevance: b. Pain management remains a major medical problem in a variety of human diseases. Chronic pain, moreover, is associated with worse disease outcome and depression. In the fullness of time, the work may allow us to determine whether the Tlx3-mediated core transcriptional program is a valid and novel therapeutic target for pain management
Project start date: 2004-01-01
Project end date: 2014-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
5R01NS047710-07 (2010): $356315
Sponsored Links Excellgen http://Excellgen.com
GENETIC CONTROL OF NOCICEPTIVE SENSORY NEURON DEVELOPMENT AND PAIN BEHAVIOR
Qiufu Ma, Associate Professor
Dana-farber Cancer Institute, 44 Binney St, Boston, Ma 02115
Grant 5R01DE018025-04 from National Institute Of Dental & Craniofacial Research
Abstract: Acute pain and chronic pain are significant complications in many human diseases, prime examples being cancer, diabetic disorders, and traumatic injury. In preliminary studies, my colleagues and I used high throughput in situ hybridization to develop a genome-scale expression map of transcription factors in developing mouse embryos. We then used this map, in combination with targeted gene disruption methods, to identify key proteins for specification of nociceptive/pain sensory neurons (nociceptors). This work led to identification of the transcription factor Runxl as a pivotal agent in development of nociceptors for thermal and neuropathic pain. The research described here builds upon this preliminary work. The goal of our research over the next five years is to define the molecular mechanisms that allow a single transcription factor (Runxl) to control the formation of a large cohort of nociceptors and the assembly of specific neural circuits for the perception of pain. We have four specific Aims. Aim 1 is to determine how Runxl expression controls the segregation of two major nociceptor subtypes, non-peptidergic versus peptidergic. Aim 2 is to address how Runxl regulates the expression of nociceptive ion channels and receptors, with the goal of understanding the logic underlying the generation of tremendous diversity within the non-peptidergic population of nociceptors. Aim 3 focuses on the assembly of pain circuits. Here we want to determine how Runxl controls the innovation of non-peptidergic nociceptors to specific peripheral and central targets. Aim 4 is to determine how distinct Runxl-dependent programs contribute to behavioral response to noxious stimuli. For each of these four aims, we have preliminary data that leads to a testable hypothesis. The predictions of these hypotheses will be tested by using a panel of genetic tools that we have already developed. In the fullness of time, the work may lead to a novel biological target and therapeutic approach for pain management
Keywords: Ablation; Acute Pain; Address; Afferent Neurons; Behavioral; Biological; Bone; Bone and Bones; Bones and Bone Tissue; Cancers; Cell Fate Control; Cell Fate Regulation; Cells; Data; Defect; Development; Disease; Disorder; Down-Regulation; Down-Regulation (Physiology); Downregulation; Embryo; Embryonic; Exhibits; Gene Down-Regulation; Gene Targeting; Gene Transcription; Generations; Genetic; Genetic Transcription; Genetics, in situ Hybridization; Genome; Goals; In Situ Hybridization; Injury; Ion Channel; Ionic Channels; Knock-in; Knock-in Mouse; Knock-out; Knockout; Label; Lead; Logic; Malignant Neoplasms; Malignant Tumor; Mammals, Mice; Maps; Mediating; Medulla Spinalis; Membrane Channels; Methods; Mice; Mice, Mutant Strains; Modality; Molecular; Murine; Mus; Mutant Strains Mice; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; Neurons; Neurons, Afferent; Neurons, Sensory; Nociception; Nociceptors; Pain; Pain Control; Pain Therapy; Pain management; Painful; Pattern; Pb element; Perception; Perinatal; Peripheral; Phenotype; Population; Programs (PT); Programs [Publication Type]; Proteins; RNA Expression; Racial Segregation; Receptor Protein; Repression; Research; Sensory; Sensory Cell Afferent Neuron; Skin; Spinal Cord; Staging; Stimulus; Targetings, Gene; Testing; Time; Transcription; Transcription Repression; Transcription, Genetic; Transcriptional Repression; Work; base; bone; chronic pain; chronic painful condition; cohort; diabetic; disease/disorder; gene product; gene repression; heavy metal Pb; heavy metal lead; human disease; in situ Hybridization Staining Method; innervation; innovate; innovation; innovative; malignancy; mouse mutant; mutant; neoplasm/cancer; nerve supply; neural circuit; neural circuitry; neuron development; neuronal; neuropathic pain; nociceptive; novel; pain behavior; painful neuropathy; postnatal; prevent; preventing; programs; prospective; receptor; response; segregation; therapeutic target; tool; transcription factor
Project start date: 2007-08-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5R01DE018025-04 (2010): $384480
5R01DE018025-03 (2009): $388363
5R01DE018025-02 (2008): $388363
1R01DE018025-01A1 (2007): $392682
MOLECULAR MECHANISMS OF FATE CHOICE IN NEURAL STEM CELLS
Qiufu Ma
Dana-farber Cancer Institute
44 Binney St
boston, Ma 02115
Grant 5P01NS047572-050003 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: A small subset of genes that encode transcription factors are expressed at spatially restricted positions and times during vertebrate central nervous system (CNS) development. These "informative" transcription factors act singly and in combinations to regulate fate choice and subtype specification. The primary goal of Project by Ma is to build a comprehensive temporal/spatial map of transcription factor expression patterns in the developing mouse CNS. Our central hypothesis is that this transcription factor atlas will identify novel regulators of early cell fate choice in neural progenitors and later maturation of specialized neurons and glia. The most carefully annotated vertebrate genome - human- encodes approximately 1,600 known or putative transcription factors of all classes (homeodomain, nuclear hormone, zinc finger, etc). In preliminary work, we have i) identified murine homologues of all human transcription factor-encoding genes, ii) designed and generated PCR primer pairs for these genes, and iii) cloned approximately 75% (1200) of them into vectors suitable for in situ hybridization (ISH). Our study plan builds upon this and other preliminary groundwork. We have three specific aims Aim 1 is to characterize the expression patterns of all transcription factor-encoding genes by ISH at key stages of forebrain, hindbrain, cerebellar and spinal cord development. Aim 2 is to catalogue the data into a searchable resource accessible to the general public via the web. The database will link ISH images to relevant annotation, accession numbers reagents, other vertebrate and invertebrate databases and human neurodevelopmental - neurodegenerative disease loci. Aim 3 is to test the central hypothesis. Towards this end, we will determine whether the atlas can be used to identify transcription factors that implement the actions of Olig and Ngn bHLH transcription factors on neuronal sub-type specification. The atlas will be scanned to identify transcription factors expressed in the Olig2 domain of spinal cord and the Ngn expression domain of the cortex. Candidate target genes will be culled by observation of aberrant expression in Olig2-/-and Ngn1/2-/- tissues, respectively and then assessed for biological function in chick neural tube assays. Aims 2 and 3 draw heavily upon the informatics and expression vector cores. The project as a whole will augment efforts in Projects by Greenberg and Stiles to identify Ngn target genes and co regulator proteins for Olig 1 and Olig2
Keywords: cell differentiation, developmental genetics, gene expression, molecular biology information system, nerve stem cell, neurogenesis, transcription factor Internet, bioinformatics, cerebellum, computational biology, developmental neurobiology, genetic regulation, mammalian embryology, molecular biology, prosencephalon, protein protein interaction, protein structure function, rhombencephalon, spinal cord chick embryo, in situ hybridization, laboratory mouse
Qiufu Ma
Dana-farber Cancer Institute
Project start date: 2004-01-01
Project end date: 2014-01-31
Sponsored Links Excellgen http://Excellgen.com