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Cerebelo como modelo genético en la sinaptogénesis

Modelos murinos para la formación y mantenimiento de conexiones entre neuronas.

Martin Radrizzani

on 9 December 2015

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Transcript of Cerebelo como modelo genético en la sinaptogénesis

Genenómica de la formación de conexiones:
Modelo Experimental: El Cerebelo Status Public on Apr 18 2001 TRANSCRIPTOME
Title Cerebellar gene xp Type non-commercial nucleotide.

Organism Mus musculus Description The Affymetrix GeneChip Mu11K was used to analyze the gene expression profile in developing mouse cerebellum (two GeneChips per E18, P7, P14, P21, and P56)
The analysis showed:
81.6% (10,321 / 12,654) of the genes represented on the GeneChip were expressed in the postnatal cerebellum, and among those,
8.7% (897 / 10,321) were differentially expressed with more than a two-fold change in their maximum and minimum expression levels during the developmental time course. Funcionamiento en red : unidades repetitivas http://www.physiol.ucl.ac.uk/research/silver_a/neuroConstruct/images/Cerebellum-1.PNG Environment, Células de Purkinje : Regulación local del crecimiento Pocos genes y muchas interacciones
Como empezar? Formacion del cerebelo Western-blot Northern Blot Differential Display “In situ” RT-PCR CPD1 b Multiple sequence alignment of members of the Anp32 family generated with ClustalW with the aid of the amino acid substitution matrix Blosum30 with a gap opening and extension penalties of 25 and 1 respectively. Alignment was visualized with the Jalview Java alignment editor. The first column shows the protein name and the species abbreviation: Hs, Homo sapiens; Cf, Canis familiaris; Mm, Mus musculus; Rn, Rattus novergicus; Xl, Xenopus laevis; Dr, Danio rerio; Rp, Rodnius prolixus; Dm, Drosophila melanogaster; Ce, Caenorhabditis elegans; Sm, Schistosoma mansoni; Pf, Plasmodium falciparum; Os, Oryza sativa; At, Arabidopsis thaliana.

Residues were coloured according to the following physicochemical criteria: blue, non-polar amino acids without G (A, F, I, L, M, P, V, W); magenta, acidic amino acids (D, E); red, basic residues without H
(R, K); green, amino acids with dissociable protons or polar side chains and G (C, G, H, N, Q, S, T, Y). Percentage indicates degree of amino acid conservation.

Low compl., low compositional complexity region;
LRR, leucine-rich repeat unit;
NES, nuclear export signal;
NLS, nuclear localisation signal.

Asterisks denote amino acid residues predicted to be on the concave surface of the LRR structure. ¿Qué los genes desregulados en los modelos de Ataxia? Modelos murinos de Ataxias:

Staggerer y weaver Anp32e/Cpd1 y PP2A en mutantes cerebelosos
staggerer y weaver
Staggerer Weaver Etapas del desarrollo del cerebelo murino Ejemplo de tinción

Antisuero contra PTEN:

Marcador de la vía NMDA

Marca en las espinas dendríticas que reciben las fibras trepadoras en células de Purkinje. Histología de PTEN durante el desarrollo Postnatal cerebelo
(DNA con Hoescht =>azul, Cpd1=>rojo)
P10 P17 Adulto IGL PcL ML IGL PcL ML EGL IGL PcL ML EGL IGL PcL ML IGL PcL ML IGL PcL ML Anp32e / Cpd1 PTEN PTEN control Weaver Staggerer PTEN


p-AKT Genes maestros Anp32e afectado en HTT Matilla and Radrizzani, The Anp32 family of proteins containing LRRs. Anp32e/Cpd1 regulates protein phosphatase 2A activity at synapses during synaptogenesis.
Costanzo RV, Vilá-Ortíz GJ, Perandones C, Carminatti H, Matilla A, Radrizzani M.
Eur J Neurosci. 2006 Jan;23(2):309-24. The Anp32 family of proteins containing leucine-rich repeats.
Matilla A, Radrizzani M.
Cerebellum. 2005;4(1):7-18. Review. Correlation between synaptogenesis and the PTEN phosphatase expression in dendrites during postnatal brain development.
Perandones C, Costanzo RV, Kowaljow V, Pivetta OH, Carminatti H, Radrizzani M.
Brain Res Mol Brain Res. 2004 Sep 10;128(1):8-19. Different conformations of phosphatase and tensin homolog, deleted on chromosome 10 (PTEN) protein within the nucleus and cytoplasm of neurons.
Moncalero VL, Costanzo RV, Perandones C, Radrizzani M.
PLoS One. 2011 Apr 29;6(4):e18857.

Downregulation of Hsp27 (HSPB1) in MCF-7 human breast cancer cells induces upregulation of PTEN.
Cayado-Gutiérrez N, Moncalero VL, Rosales EM, Berón W, Salvatierra EE, Alvarez-Olmedo D, Radrizzani M, Ciocca DR.
Cell Stress Chaperones. 2012 Aug 21. Lack of interaction between ANP32E, PP2A and huntingtin in neuritic aggregates of Human affected by the Huntington disease.
Perandones C, Costanzo RV, Radrizzani M.
Manuscript under revision. The rate of Tau synthesis is differentially regulated during postnatal development in mouse cerebellum.
Vilá-Ortiz GJ, Santa-Coloma TA, Carminatti H, Radrizzani M.
Cell Mol Neurobiol. 2001 Oct;21(5):535-43. Differential expression of CPD1 during postnatal development in the mouse cerebellum.
Radrizzani M, Vilá-Ortiz G, Cafferata EG, Di Tella MC, González-Guerrico A, Perandones C, Pivetta OH, Carminatti H, Idoyaga Vargas VP, Santa-Coloma TA.
Brain Res. 2001 Jul 13;907(1-2):162-74. Developmental regulation of Thy 1.2 rate of synthesis in the mouse cerebellum.
Radrizzani M, Carminatti H, Pivetta OH, Idoyaga Vargas VP.
J Neurosci Res. 1995 Oct 1;42(2):220-7. Sintesis de proteínas vs fosforilación PTEN y Autismo Behav Neurosci. 1994 Oct;108(5):848-65.
Impairment in shifting attention in autistic and cerebellar patients.
Courchesne E, Townsend J, Akshoomoff NA, Saitoh O, Yeung-Courchesne R, Lincoln AJ, James HE, Haas RH, Schreibman L, Lau L. Association of upregulated Ras/Raf/ERK1/2 signaling with autism.
Zou H, Yu Y, Sheikh AM, Malik M, Yang K, Wen G, Chadman KK, Brown WT, Li X.
Genes Brain Behav. 2011 Jul;10(5):615-24. doi: 10.1111/j.1601-183X.2011.00702.x. Epub 2011 Jun 14. Pten regulates neuronal arborization and social interaction in mice.
Kwon CH, Luikart BW, Powell CM, Zhou J, Matheny SA, Zhang W, Li Y, Baker SJ, Parada LF.
Neuron. 2006 May 4;50(3):377-88. Anp32e y Autismo Autistic-like behaviour and cerebellar dysfunction in Purkinje cell Tsc1 mutant mice.
Tsai PT, Hull C, Chu Y, Greene-Colozzi E, Sadowski AR, Leech JM, Steinberg J, Crawley JN, Regehr WG, Sahin M.
Nature. 2012 Jul 1. doi: 10.1038/nature11310. [Epub ahead of print] Muchas gracias por su atención Asociación entre Anp32e/PP2A a PTEN? van Diepen MT, Parsons M, Downes CP, Leslie NR, Hindges R, Eickholt BJ. Nat Cell Biol. 2009 Oct;11(10):1191-6. Epub 2009 Sep 20. MyosinV controls PTEN function and neuronal cell size. Expresion diferencial
en cerebelo y tejidos Expresion diferencial
en el cerebelo durante el desarrollo. PP2A ANP32e CK2
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