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pain in weaned rats.

doi:10.1016/j.ijdevneu.2012.03.289

58 P.P. Kale, V. Addepalli / Int. J. D

he dopaminergic stabilizer, (−)-OSU6162, rescues striataleurons with normal and expanded polyglutamine chains

n huntingtin protein from exposure to free radicals anditochondrial toxins

. Ruiz 1,2, M.J. Casarejos 2,3, M.A. Mena 2,3,∗, J.G. de Yebenes 1,2

Department of Neurology, Hospital Ramón y Cajal, SpainCIBERNED, Instituto de Salud Carlos III, SpainDepartment of Neurobiology, Hospital Ramón y Cajal, Madrid, Spain

Huntington’s disease (HD) is a neurodegenerative disease char-cterized by progressive motor, cognitive and psychiatric deficits,ssociated with predominant loss of striatal neurons and caused bypolyglutamine expansion in the huntingtin protein. There is so fareither cure nor approved disease-slowing therapy for HD, thoughecent clinical studies have shown a beneficial long-term effect ofridopidine in patients with HD. The nature of this effect, purelyymptomatic or, in addition, neuroprotective, is difficult to eluci-ate in clinical trials. Pridopidine and (−)-OSU6162 are members ofnew family of compounds referred to as dopaminergic stabilizers,hich normalize abnormal dopamine neurotransmission.

We investigated the effects of (−)-OSU6162 on huntingtinnocked-in striatal neurons in culture. Control neurons had normalull-length huntingtin with 7 glutamines while “mutant” neuronsad large expansions (Q = 111). We studied the dose-effect curvesf (−)-OSU6162 on mitochondrial activity, LDH levels, necrosisnd apoptosis in untreated Q7 and Q111 cells. In addition, wenvestigated the effects of (−)-OSU6162 on Q7 and Q111 neuronshallenged with different neurotoxins such as sodium glutamate,2O2, rotenone and 3-nitropropionic acid (3NP). As we found pre-ention of toxicity of some of these neurotoxins, we investigatedhe putative neuroprotective mechanisms of action of (−)-OSU6162

easuring the effects of this dopaminergic stabilizer on the ratiosf Bcl2/Bax proteins and of p-ERK/ERK, the levels of chaperonesnd GSH, and the effects of (−)-OSU6162 on dopamine uptake andelease.

We found that (−)-OSU6162, 3-150 �M, produces a dose depen-ent increase of mitochondrial activity and a reduction of celleath. (−)-OSU6162 does not change glutamate toxicity, but it par-ially prevents that of H2O2, rotenone and 3-nitropropionic acid.−)-OSU6162 increases the levels Bcl2/Bax and decreases thosef p-ERK/ERK and CHIP in Q111 cells. (−)-OSU6162 increased 3H-opamine uptake and amphetamine-induced 3H-dopamine release

n E13 mouse mid brain neurons. Our studies demonstrate that (−)-SU6162 improves survival and mitochondrial function in striatal111 neurons and the resistance of these cells to several neurotox-

ns that damage striatal neurons, suggesting that (−)-OSU6162 andelated compounds should be tested for neuroprotection in animalodels and, eventually, in patients with HD.

oi:10.1016/j.ijdevneu.2012.03.287

sing molecular signals controlling embryonic development toirect the differentiation of corticospinal motor neurons

oushan Melissa Lin 1,∗, William Hendriks 2, Paola Arlotta 1

Department of Stem Cell and Regenerative Biology and Harvard Stemell Institute, Harvard University, United StatesHarvard Stem Cell Institute iPS Core Facility, Harvard University,nited States

The molecular signals that govern the development of distinct

uroscience 30 (2012) 640–671

vant neuronal population that degenerates in Amyotrophic LateralSclerosis (ALS), and are permanently damaged in spinal cord injury(SCI).

With the identification of a series of neuronal subtype-specificgenes that mark CSMN and other subcerebral projection neurons(SCPN) in vivo, it was notably discovered that the transcriptionfactor Forebrain Embryonic Zinc Finger 2 (Fezf2) is necessary forthe birth and early differentiation of all SCPN, including CSMN(i.e. Fezf2 −/− mice lack SCPN in their cortex), and is sufficient to“fate-switch” cortical progenitors fated to form upper layer callosalneurons to generate deep layer corticofugal projection neurons(CfuPN) that also includes CSMN.

The central role played by Fezf2 in early SCPN differentiation ledus to investigate whether cell-autonomous developmental signalsthat direct early SCPN development in the embryo could be usedto selectively generate these neurons, from dorsal telencephalicprogenitors derived from mouse induced pluripotent stem (miPS)cells, in vitro. We are currently testing whether overexpression ofFezf2 and other selected genes can progressively instruct the neu-ron type-specific differentiation of miPS cells into CfuPN, includingCSMN.

doi:10.1016/j.ijdevneu.2012.03.288

Nociceptive behaviour and lumbar spinal NADPH-d/NOS, p-ERKAND c-FOS expression after formalin pain induction in weanedrats

Gila Behzadi

Shahid Beheshti Med. Sci. University, Faculty of Medicine,Neuroscience Res. Ctr. And Physiology Department, Tehran, Iran

Introduction: The sensitivity to formalin pain is an expressionof neuronal plasticity in pain processing as the nervous systemdevelops. Upon our previous study, the formalin pain responsive-ness showed the complete appearance of phase II in 21 day-oldweaned rats. In the present study we therefore were investigatingthe expression of Nitric Oxide Synthase by NADPH-d histochem-istry and nNOS immunoreaction along with the p-ERK and c-FOSexpression in lumbar spinal cord.

Methods: Following right intraplantar injection of formalin solu-tion (%10), coronal frozen section from L4–L5 were processed foreach histochemical purposes.

Results: 1–90 min after formalin injection we observed %25 and%30 increase in the number of NADPH-d and NOS containing neu-rons in lamina I-II of dorsal horn (p < 0.01). 2–5 min after formalininjection the number of p-ERK positive neurons were increased inlamina I-II significantly (p < 0.001). 3–120 min after formalin injec-tion extensive number of c-FOS protein was expressed in laminaI–II neurons (p < 0.001).

Discussion: Our results demonstrate a mature neuronal activa-tion of lumbar spinal dorsal horn during phases I and II of formalin