Mice with disrupted midsized and heavy neurofilament genes lack axonal neurofilaments but have unaltered numbers of axonal microtubules.
Mammalian neurofilaments are assembled from the light (NF-L), midsized (NF-M), and heavy (NF-H) neurofilament proteins. While NF-M and NF-H cannot self-assemble into homopolymers, the data concerning NF-L has been more contradictory. In vitro bovine, porcine, and murine NF-L can homopolymerize in the absence of other subunits. However, in vivo studies suggest that neither rat nor mouse NF-L can form filaments when transfected alone into cells lacking endogenous intermediate filaments. By contrast, human NF-L forms homopolymers in similar cell lines. Recently we generated mice with null mutations in the NF-M and NF-H genes. To determine if mouse NF-L can homopolymerize in mouse axons, NF-M and NF-H null mutants were bred to create a line of double mutant animals. Here we show that axons in NF-M/H double mutant animals are largely devoid of 10-nm filaments. Instead, the axoplasm is transformed to a microtubule-based cytoskeleton-although the lack of any increase in tubulin levels per unit length of nerve or of increases in microtubule numbers relative to myelin sheath thickness argues that microtubules are not increased in response to the loss of neurofilaments. Thus in vivo rodent neurofilaments are obligate heteropolymers requiring NF-L plus either NF-M or NF-H to form a filamentous network.
Department of Psychiatry, Mount Sinai School of Medicine, New York, New York 10029, USA.
G A
GA
Elder
V L
VL
Friedrich
D
D
Pereira
P H
PH
Tu
B
B
Zhang
V M
VM
Lee
R A
RA
Lazzarini
D000818:Animals; D001369:Axons / Q000378:metabolism / Q000648:ultrastructure*; D001704:Biopolymers; D015153:Blotting, Western; D007382:Intermediate Filaments / Q000648:ultrastructure*; D051379:Mice; D008817:Mice, Mutant Strains; D008854:Microscopy, Electron; D008870:Microtubules / Q000648:ultrastructure*; D016900:Neurofilament Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
D001704:Biopolymers; D016900:Neurofilament Proteins; D012333:RNA, Messenger; D014404:Tubulin