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[P105]: Study of classical neurotransmitters in larval and postmetamorphic retinas of the sea lamprey
Author(s) -
VillarCerviño V.,
Abalo X.,
BarreiroIglesias A.,
Holstein G.R.,
Martinelli G.,
Rodicio C.,
Anadón R.
Publication year - 2006
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/j.ijdevneu.2006.09.167
Subject(s) - humanities , mount , cartography , geography , art , computer science , operating system
A better understanding of the molecules that control midbrain dopaminergic neuron (mesDA) fate specification would greatly facilitate the establishment of novel paradigms for stem cell differentiation and thus the development of therapeutic strategies for drug screen and cell therapy for treating Parkinson’s disease. We have generated Pitx3-GFP knock-in mice that express an eGFP reporter specifically in developing and adult substantia nigra DA neurons. The heterozygous Pitx3-GFP mice exhibit normal midbrain dopaminergic development and thus provide us with a unique opportunity for isolating primary nigral DA neurons for analysing gene expression patterns associated with this specific cell population. We have performed transcriptome profiling of embryonic (E) day 12 midbrain DA (Pitx3-GFP) and non-DA (Pitx3-GFP−) mesencephalic cells by Affymetrix microarray analysis using fluorescence activated cell sorter (FACS) purified cells. In mice, neurogenesis is almost complete at E12 but gliogenesis has not yet begun so the Pitx3-GFP− population contains mostly neuronal cells of non-DA character with a small fraction of neural progenitor/stem cells. Therefore, the difference in expression profile between the two groups is likely to derive primarily from genes that determine neuronal subtypes (e.g. DAergic or nonDAergic). Furthermore, this screen provides information for the presence or absence of transcripts coding for growth factors, their receptors and other signalling molecules in the E12 ventral midbrain (VM). Our microarray data revealed that all known midbrain DA specific transcripts are robustly differentially detected in the Pitx3-GFP population. Therefore, this screen serve as a solid platform to identify factors that are capable of promoting midbrain DA fate and/or functional maintenance of midbrain DA neurons. We are currently validating the expression of selected candidate genes (to be referred to as mda1, 2, etc.) by in situ hybridisation in brain sections. The poster will be focused on reporting midbrain expression pattern of the mda genes validated so far.