Cultivation and characterization of planarian neuronal cells isolated by fluorescence activated cell sorting (FACS).
Studies using molecular markers have revealed that planarians possess a highly organized brain. Here we separated brain neurons from dissociated planarian head cells by fluorescence activated cell sorting (FACS), and characterized them by single cell PCR analysis and cell culture. Dissociated cells were labeled with three different fluorescent dyes, Hoechst 33258, Merocyanine 540, and Propidium Iodide (PI), and fractioned by FACS. Interestingly, we have succeeded in identifying a cell fraction specific to the head, which we have named the head-abundant cell fraction (HAC). Most of the HAC expressed neuron-specific genes and proteins. When they were cultured in vitro, they showed an ability to extend neurites on several types of extracellular matrices (ECMs), and, depending on the ECM type used, presented a high level of plasticity in morphology and gene expression.
Asami
Maki
M
Department of Biofunctional Chemistry, Faculty of Biomolecular Science, Okayama University Graduate School of Natural Science and Technology, Okayama, Japan.
Nakatsuka
Tsuyoshi
T
Hayashi
Tetsutaro
T
Kou
Kenji
K
Kagawa
Hiroaki
H
Agata
Kiyokazu
K
D000818:Animals; D001921:Brain / Q000166:cytology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation; D006257:Head; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D009928:Organ Specificity; D010932:Planarians / Q000033:anatomy & histology / Q000166:cytology*; D016133:Polymerase Chain Reaction
D005456:Fluorescent Dyes