Synaptic integrins in developing, adult, and mutant muscle: selective association of alpha1, alpha7A, and alpha7B integrins with the neuromuscular junction.
Differentiation of both pre- and postsynaptic structures at the skeletal neuromuscular junction is organized by the basal lamina that occupies the synaptic cleft. As beta1 integrins are a major class of receptors for basal lamina components, we stained muscles with antibodies to the 10 integrin alpha subunits known to form dimers with beta1, to determine if any of these molecules were concentrated at synaptic sites on muscle fibers. In both developing and adult muscle, the integrin alpha1 chain was selectively associated with presynaptic cells (Schwann cells and/or nerve terminals), while alpha7 was present on both synaptic and extrasynaptic portions of the muscle fiber surface. Thus alpha1 and alpha7 integrins are present in synaptic membranes. Expression of the alpha7 chain was analyzed further by staining with antibodies specific for three alternatively spliced products of the alpha7 gene (A, B, and C), all of which were expressed in muscle. The alpha7A and alpha7B isoforms were confined to synaptic sites in adult muscle, while alpha7C was present both synaptically and extrasynaptically. In developing muscle, alpha7A appeared postnatally and specifically at the synapse; alpha7B was present throughout the muscle fiber perinatally, becoming confined to the synapse in the second postnatal week; and alpha7C was present extrasynaptically both perinatally and in adulthood. Thus, two of the alpha7 integrins are synapse-specific, and all three show distinct spatiotemporal patterns of expression within a single cell type. Finally, we asked whether perturbation of laminin expression affected the distribution of the alpha7 integrins. In normal mice, laminin beta2 is concentrated in synaptic basal lamina. In beta2-null mutant mice, alpha7A was still present at synaptic sites, but alpha7B was absent. This result provides genetic evidence that basal lamina composition is a determinant of integrin distribution.
Martin
P T
PT
Department of Anatomy and Neurobiology, Washington University Medical School, St. Louis, Missouri 63110, USA.
Kaufman
S J
SJ
Kramer
R H
RH
Sanes
J R
JR
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002478:Cells, Cultured; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D016023:Integrins / Q000502:physiology*; D007797:Laminin / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009469:Neuromuscular Junction / Q000378:metabolism*; D010525:Peripheral Nerves / Q000378:metabolism; D051381:Rats; D013570:Synaptic Membranes / Q000378:metabolism*
D016023:Integrins; D007797:Laminin