1 / 16

Nikolay N . Dygalo

Functional neurogenomics : brain neurochemical systems in regulation of psychoneuroendocrine functions. Nikolay N . Dygalo. Institute of Cytology & Genetics, RAS, Novosibirsk. International Workshop EC-Russia: Cooperation in Medicine and Public Health St.-Petersburg 11-12 October, 2007.

evelyn
Télécharger la présentation

Nikolay N . Dygalo

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Functional neurogenomics: brain neurochemical systems in regulation of psychoneuroendocrine functions Nikolay N. Dygalo Institute of Cytology & Genetics, RAS, Novosibirsk International WorkshopEC-Russia: Cooperation in Medicine and Public Health St.-Petersburg 11-12 October, 2007

  2. Experimental Model of Neurodegeneration (natural apoptotic cell death in the week-old rat brain) DNA fragmentation Caspase-3 protein Cortex Brainstem Cerebellum Hippocamp. Bannova et al., Developmental Brain Res. 2004 Menshanov, Bannova, Dygalo, Neurochem. Res. 2006

  3. Alpha2-adrenergic receptors (α2-ARs) regulate apoptosis in the cortex Antisense knock-down of α2A-ARs increased caspase-3 expression in the week-old rat cortex and α2-AR agonist clonidine decreased it. Data evidence for neuroprotective potency of α2-AR agonists in the cortex.

  4. Control Yohimbine Clonidine Clonidine + Yohimbine 180 160 140 120 caspase-3 mRNA,% 100 80 , 60 40 20 0 Stimulation of 2-ARincreases caspase-3 mRNA in the week old rat brainstem caspase-3 mRNA β-actin mRNA Clonidine + Control Yohimbine Yohimbine Clonidine - Antagonist - Agonist Dygalo N.N. et al., Developmental Brain Res. 2004. Menshanov PN, Bannova AV, Dygalo NN. Neurochem Res. 2006.

  5. RNA interference: Sequence-specific inhibition of the gene expression in the brain in vivo 5-day-old rat brain Oligonucleotides (ОDN) enter brain cells Site of injection Hematoxylin & Eosin Green fluorescence- ОDN

  6. Saline dcRNA siRNA 5-day-old rat brain siRNA inhibited targeted-gene expressionin the brain in vivo Site of injection of siRNAtargetedα2A-adrenergic receptor (α2A-AR) gene α2А-АR mRNA -actin mRNA Substances injected into the brain Shishkina G.T., Kalinina T.S., Dygalo N.N. Neuroscience, 2004.

  7. RNA interference(targeting2A-ARmRNA) blocks behavioral response of the animal to the receptor stimulation 600 Saline dsRNA siRNA 480 360 Time to the first paw movement, sec * 240 120 0 0 5 10 Clonidine, mkg F2,55=15.82; p<0.001

  8. Up-regulation of tryptophan hydroxylase-2 mRNA in the rat brain by chronic fluoxetine treatment correlates with its antidepressant effect Immobility in the forced-swim test TPH2 mRNA Correlation immobility - TPH2 mRNA Real-time PCR Tissue sample r = -0.485, p<0.05 Shishkina, Kalinina, Dygalo, Neuroscience, 2007

  9. Antidepressant Sensitive Catalepsy (ASC) mouse strain with depressive-like features * * Distance run, cm AKR CBA ASC ASC mice have decreased open field locomotion * Catalepsy in ASC mice is sensitive to chronic, but not acute, treatment with classic antidepressant drugs, imipramine and fluoxetine. Immobility time, s AKR CBA ASC ASC mice have increased immobility in the forced swim test ASC – originated from backcrossesbetweenCBA и AKR mouse strains Bazovkina et al., Genetika, 2005 Tikhonova et al., Bull. Exp. Biol. Med., 2006

  10. Linkage between the main gene of catalepsy and 5-HT1A serotonin receptor in mice QTL analysis mapped the main gene of catalepsy on the 50-75 cM fragment of mouse chromosome 13 near the 5-HT1A receptor gene Selective breeding of mice for catalepsy removes the AKR-derived marker linked to 5-HT1A receptor and fixed the CBA-like. Kondaurova et al., Genes Brain Behav., 2006 Kulikov et al., Dokl. Acad. Sci. 2003, Genetika, 2003

  11. C1473G polymorphism in mouse tph2 gene (tryptophan hydroxylase-2) determines the activity of this key enzyme of serotonin synthesis in the brain and intermale aggressiveness in mice C1473G polymorphism in mouse tph2 gene Tph2 activity Number of attacks Strains: 1-CBA, 2-PT, 3-C57BL/6, 4-C3H/He, 5-AKR, 6-YT, 7-DD, 8-BALB/c, 9-A/He, 10-CC57BR, 11-DBA/2 Kulikov et al., Genes Brain Behav., 2005

  12. Sequencing of the 5-HT1A serotonin receptor gene of hibernating ground squirrel revealed insertion of three nucleotides (marked in red) in the third intracellular loop that is lacking in the 5-HT1A receptor gene of nonhibernators Naumenko et al., Genes Brain and Behavior, 2007

  13. base level • water loading • water loading + 5-HTP injection Vasopressin deficient Brattboro rats were used to analyze interrelations between serotonin and vasopressin in regulation of renal function Vasopressin deficiency affects brain 5-HT system reactions to hydration and water deprivation. Serotonin has a role in regulation of renal function. Activity of TPH2 (pmol/mg/min) Glomerular filtration rate • control • water deprivation • hydration WISTAR BRATTLBORO Ivanova et al. Neuroendocrinology, 2007 Popova et al. Pflugers Arch., 2002

  14. Models: Experimental Model of Neurodegeneration Genetic Models of Psychopathology Genetic Model of Vasopressin Deficiency Natural Model of specific form of behavior (hibernation) Depression-like behavior Methods: Gene silencing: RNA interference, Antisense knockdown Gene expression: Real-Time PCR, RT-PCR, Westernblot Gene sequencing Quantitative trait linkage analysis Neurochemistry: Enzymes activities, Receptors densities, Neurotransmitters concentrations (HPLC) Behavioral tests Measurements of physiological functions Applications: Neurodegeneration Psychopathology Target validation for new drugs Search for new targets of the existing drugs

  15. Participants of the work: Institute of Cytology & Genetics, RAS, Novosibirsk Laboratory of Functional Neurogenomics Laboratory of Behavioral Neurogenomics Laboratory of Physiological Genetics Institute of Chemical Biology & Fundamental Medicine, RAS, Novosibirsk

  16. Thank you for attention

More Related