A reliable in vitro model for studying peripheral nerve myelination in mouse.
The rat dorsal root ganglia (DRG) model is a long-standing in vitro model for analysis of myelination in the peripheral nervous system. For performing systematic, high throughput analysis with transgenic animals, a simplified BL6 mouse protocol is indispensable. Here we present a stable and reliable protocol for myelinating co-cultures producing a high myelin ratio using cells from C57BL/6 mice. As an easy accessible and operable method, Sudan staining proved to be efficient in myelin detection for fixed cultures. Green fatty acid stain turned out to be highly reliable for analysis of the dynamic biological processes of myelination in vital cultures. Once myelinated we were able to induce demyelination by the addition of forskolin into the model system. In addition, we provide an optimised rat DRG protocol with significantly improved myelin ratio and a comparison of the protocols presented. Our results strengthen the value of ex vivo myelination models in neurobiology.
Department of Neurology, Medical Faculty, Research Group for Clinical and Experimental Neuroimmunology, Heinrich-Heine-University, Düsseldorf, Germany. mark.stettner@med.uni-duesseldorf.de
Mark
M
Stettner
Kathleen
K
Wolffram
Anne K
AK
Mausberg
Christian
C
Wolf
Sebastian
S
Heikaus
Angelika
A
Derksen
Thomas
T
Dehmel
Bernd C
BC
Kieseier
D000818:Animals; D001391:Azo Compounds; D015153:Blotting, Western; D001896:Boron Compounds; D002478:Cells, Cultured / Q000502:physiology; D005576:Colforsin / Q000494:pharmacology; D004396:Coloring Agents; D003711:Demyelinating Diseases; D005456:Fluorescent Dyes; D005727:Ganglia, Spinal / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D004676:Myelin Basic Protein / Q000096:biosynthesis / Q000235:genetics; D018993:Myelin P0 Protein / Q000096:biosynthesis / Q000235:genetics; D009186:Myelin Sheath / Q000502:physiology* / Q000648:ultrastructure; D009281:Naphthalenes; D010169:Palmitic Acids; D061251:Primary Cell Culture; D051381:Rats; D011917:Rats, Inbred Lew; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D012583:Schwann Cells / Q000378:metabolism; D012584:Sciatic Nerve / Q000166:cytology; D013045:Species Specificity; D013194:Staining and Labeling / Q000379:methods
C543305:4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-hexadecanoic acid; D001391:Azo Compounds; D001896:Boron Compounds; D004396:Coloring Agents; D005456:Fluorescent Dyes; D004676:Myelin Basic Protein; D018993:Myelin P0 Protein; D009281:Naphthalenes; D010169:Palmitic Acids; D005576:Colforsin; C016118:Sudan Black B