Cardiotrophin-1 requires LIFRbeta to promote survival of mouse motoneurons purified by a novel technique.
The cytokines ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF) signal through a receptor complex formed between two transmembrane proteins, gp130 and LIFRbeta. In addition, CNTF also uses a ligand-binding component which is anchored to the cell membrane. In the case of cardiotrophin-1 (CT-1), LIFRbeta is also required in cardiomyocytes, but this has not been proven in neurons, and published data suggest that motoneurons may use a different receptor complex. We used Lifrbeta knockout mice to assess the requirement for this receptor component in the signal transduction of CT-1 in motoneurons. To study purified motoneurons from such mutants, we have developed a method allowing for isolation of highly purified mouse motoneurons. This protocol is based on the immunoaffinity purification of motoneurons using antibodies against the extracellular domain of the neurotrophin receptor, p75, followed by cell sorting using magnetic microbeads. We show that CNTF, LIF, and CT-1 are unable to promote the survival of motoneurons derived from homozygous Lifrbeta-/- mutant embryos. Thus, LIFRbeta is absolutely required to transduce the CT-1 survival signal in motoneurons.
Arce
V
V
INSERM U.382, Institut de Biologie du Développement de Marseille (INSERM-CNRS-Université de la Méditerranée-AP de Marseille), Campus de Luminy, France.
Garces
A
A
de Bovis
B
B
Filippi
P
P
Henderson
C
C
Pettmann
B
B
deLapeyrière
O
O
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation; D002470:Cell Survival / Q000502:physiology; D016207:Cytokines / Q000502:physiology*; D006131:Growth Inhibitors / Q000502:physiology*; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D009046:Motor Neurons / Q000166:cytology* / Q000378:metabolism; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D016207:Cytokines; D006131:Growth Inhibitors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor; C092073:cardiotrophin 1