Molecular characteristics of in vitro differentiated dopaminergic neurons

Zhigang GONG; Rongjun ZHANG; Xiaodong WANG; Qing LAN

ABSTRACT

BACKGROUND:

A series of studies have demonstrated that ventral mesencephalic dopamine precursors may differentiate into considerable dopaminergic neurons (DNs) available for cell transplantation for Parkinson's disease.However,the detailed molecular characteristics of these cells remain undear.

OBJECTIVE:

To observe the expression of main marker genes of DNs cultured in vitro.DESIGN,TIME AND

SETTING:

This observation experiment was performed at the Brain Tumor Laboratory in the Second Affiliated Hospital of Soochow University and Shanghai Biochip Company from December 2005 to December 2008.MATERIALSInbred pregnant Wistar rats (F+36) weighing 300 g approximately were used for in vitro culture of mesencephalic cells.

METHODS:

Ventral mesencephalic cells from inbred pregnant Wistar rats were cultured with basic fibroblast growth factor,and were induced to differentiate in vitro with L-Ascorbic acid-2-phosphate sesquimagnesium salt.Total RNA were extracted before and 4,7 days after the induction.Affymetrix microarrays were utilized to detect the expression changes of main marker genes of DNs.MAIN OUTCOME

MEASURES:

Morphologic changes of differentiated ventral mesencephalic precursors;Expression of DN-ralated common marker genes before and after the induced differentiation.

RESULTS:

Rat ventral mesencephalic precursors differentiated into numerous tyrosine hydroxylase-positive cells,which expressed multiple marker genes of DNs,such as AHD2,calcium binding protein 1 (Cab1),DDC,DAT,Drd2,Girk2,VMAT-2,with variously degrees and with different tendencies.Among those genes,only Cab1 was significantly up-regulated and only Girk2 was significantiy down-regulated.

CONCLUSION:

Rat ventral mesencephalic precursors can be induced to differentiate into plenty of tyrosine hydroxylase-positive DNs,mainly originating in A10 regions.