research group of functional genomics and proteomics
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Research group of Functional Genomics and Proteomics
• - 9 postdocs• - 11 PhD students• - 1 IOF manager• - 4 technicians
http://bio.kuleuven.be/df/ls/people
Research group of Functional Genomics and Proteomics
Human brain ~100 billion
neurons
Caenorhabditis elegans 302 neurons
Neuropeptides are key regulators of animal physiology and behaviour through activation of GPCRs.
Conserved mechanisms: reproduction, locomotion, learning, digestion, ageing…
L
L
Neuropeptides, neuronal communication and neuroplasticity
Big ideas from small brains
• Learning, memory, and ageing• Cancer• Feeding behavior• Reproduction• Sleeping disease
Main research topics:
Characterization of novel neuropeptide-mediated signaling systems in C. elegans
Screening for GPCR ligand
Expression analysis
Behavioral assays
e.g. locomotion RNAi knockdown
Neuropeptides are key regulators of animal physiology (feeding, reproduction) through binding of GPCRs.
C. elegans: neurobiological model
Contact:Contact: Isabel.beets@bio.kuleuven.beIsabel.beets@bio.kuleuven.be
Caenorhabditis elegans advantages:302 neuronen, toolbox for genetic manipulationsthousands of mutant strains available
The locomotive behaviour of the worm can be manipulated by light by making transgene worms in which specific neurons express light-sensitive ion channels, Channelrhodopsine-2 (ChR2, depolarisation, activation) and Halorhodopsine.
Photoactivation of defined neurons in the head let the worms crawl in the shape of a triangleImage modified from Stirman et al. (2011) Nature Methods 8: 153-158
Understanding experience-based learning behaviour: The unraveling of neuronal circuits by innovative optogenetic tools.
Aim: can we control learning behaviour using optogenetics?
A forward mutagenesis screen to study the neuroendocrine regulatory system of reproduction in C. elegans
Mutagenesis
GFP expression in intestine
Fluorescent worms
(vit-2::GFP)COPAS worm sorter
Non fluorescent worms
Gene mappingOR
Whole genome sequencing
Target phenotypingDefects with respect to
reproductionE.g. egg-laying defect
Proteomics and peptidomics of long-lived Proteomics and peptidomics of long-lived C. elegansC. elegans
Techniques
Many life-extending interventions available in C. elegans→ insulin pathway mutations, dietary restriction,…
Studying long-lived worms to help us combat aging-associated diseases→ interventions that increase lifespan also decrease incidence of cancer, neurodegenerative diseases,…
PeptidomicsDifferential labeling of peptides Liquid chromatographyMass spectrometry
ProteomicsGel electrophoresis (2D-DIGE)Mass spectrometry
What are the effectors What are the effectors that control aging?that control aging?
How do the endocrine How do the endocrine systems influence aging?systems influence aging?
Contact:Contact: Wouter.dehaes@bio.kuleuven.beWouter.dehaes@bio.kuleuven.be
GnRHR achtige receptoren bij C. elegans: Reproductie of energiehuishouding?
CONTACT:Lotte.frooninckx@bio.kuleuven.beResearch group of Functional Genomics and Proteomics
VERTEBRATENReceptor wordt geactiveerd door het gonadotropin-releasing hormoon (GnRH) dat de reproductie reguleert.
INSECTENReceptor wordt geactiveerd
door adipokinetisch hormoon (AKH)
dat de energiehuishouding reguleert. C. elegans
8 homologen: gnrr-1 tot gnrr-8Waar komen deze genen tot
expressie?GFP-lokalisatie, PCR, microscopie
Wat is de invloed van een deletie in deze genen: fenotypering, microscopie,microfluidica
Wat zijn de liganden?‘ontwezen receptoren’, celculturen, high-throuput screenen
Neuropeptide hormones and their receptors: a suitable target to combat tsetse flies?
Tsetse flies: vectors for parasites that cause sleeping disease in human and nagana in cattle. There is an urgent call for the development of new species-specific insecticides.G protein-coupled receptors are promising targets for the development of insecticides based on peptidomimetica.
The tsetse fly genome has been sequenced recently and a large number of putative neuropeptides and GPCRs are predicted. The goal is to deorphanize some GPCRs, study their tissue distribution and investigate the binding characteristics of the neuropeptides.
Techniques: - Molecular biology RNA/DNA purification, PCR, sequencing, Reverse pharmacology, Quantitative real-time PCR, Structure-activity studies
Promotor: Prof. Dr. L. Schoofs (http://bio.kuleuven.be/df/LS/)Mentor: Jelle Caers (contact: Jelle.Caers@bio.kuleuven.be)
2D-DIGE2D-DIGE
Differential Differential proteomicsproteomics
Microarray-Microarray-analysisanalysis
TranscTranscript-ript-
omicsomicsMass Mass
spectrometryspectrometry
PeptidomicsPeptidomics
RNA RNA interferenceinterference
Gene SilencingGene Silencing
gregarious solitary
Phenotypic plasticity of physiology and behaviour
Next Next generation generation sequencingsequencing
Elucidation of locust phase polyphenism with gel based proteomics.
2 phenotypes
1 Genome
Epigenetic component?
DNAMethyl-Cytidine
Differential DNA methylation?
Differential PROTEOME?
Workflow:
Tissue dissection
Protein extraction
Differential analysis: 2D-DIGE
Protein identification- MALDI TOF/TOF mass spectrometry- Bioinformatics de novo protein identification
Brain
Bart.Boerjan@bio.kuleuven.be
More info?
Towards a novel insect pest control strategy
More info: Bart.Boerjan@bio.kuleuven.be
Creativity
Bioinformatics
Genetic engineering
confidential
Extensive training in:-proteomics-peptidomics-cell culture-scientific reasoning(SDS-PAGE, 2D-DIGE, mass spectrometry)
Royality in honeybees-the making of a queen
Uli.Ernst@bio.kuleuven.be
Royal jelly, active (protein)
compound: royalactin, 57kDa
royalactin: activates epidermal growth factor
receptor (Egfr) in the fat body
-Epigenetic changes-High juvenile hormone titer-...
...but: how can a 57kDa protein reach the fat body
via the gut???
Annona Cherimoya & cancer
Septic shock
Lethal dose
Sub-Lethal dose
Protected against
Identification of protective factor
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