Diminished climbing fiber innervation of Purkinje cells in the cerebellum of myosin Va mutant mice and rats.
Myosin Va is an actin-based molecular motor that is involved in organelle transport and membrane trafficking. Here, we explored the role of myosin Va in the formation of synaptic circuitry by examining climbing fiber (CF) innervation of Purkinje cells (PCs) in the cerebella of dilute-neurological (d-n) mice and dilute-opisthotonus (dop) rats that have mutations in dilute-encoded myosin Va. Anterograde labeling of CFs with biotinylated dextran amine (BDA) revealed that they arborized poorly and that their tips extended only half way through the thickness of the molecular layer (ML) in adult d-n mice. Using immunohistochemistry specific for vesicular glutamate transporter 2 (VGluT2) to visualize CF synaptic terminals, we found that during development and in adulthood, these terminals did not ascend as far along the proximal shaft dendrites of PCs in d-n mice and dop rats as they did in normal animals. An irregular distribution of BDA-labeled bulbous varicosities and VGluT2 spots along CF branches were also noted in these animals. Finally, VGluT2-positive CF terminals were occasionally localized on the PC somata of adult d-n cerebella. These phenotypes are consistent with our electrophysiological findings that CF-mediated excitatory postsynaptic currents (EPSCs) were significantly smaller in amplitude and faster in decay in adult d-n mice, and that the regression of multiple CFs was slightly delayed in developing d-n mice. Taken together, our results suggest that myosin Va is essential for terminal CF extension and for the establishment of CF synapses within the proper dendritic territories of PCs.
Takagishi
Yoshiko
Y
Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-8601, Japan. taka@riem.nagoya-u.ac.jp
Hashimoto
Kouichi
K
Kayahara
Tetsuro
T
Watanabe
Masahiko
M
Otsuka
Hiroyuki
H
Mizoguchi
Akira
A
Kano
Masanobu
M
Murata
Yoshiharu
Y
D000344:Afferent Pathways / Q000002:abnormalities* / Q000166:cytology / Q000378:metabolism; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives; D020013:Calcium Signaling / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics; D002525:Cerebellar Cortex / Q000002:abnormalities* / Q000166:cytology / Q000378:metabolism; D003911:Dextrans; D019706:Excitatory Postsynaptic Potentials / Q000235:genetics; D005260:Female; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D046529:Microscopy, Electron, Transmission; D018995:Myosin Heavy Chains / Q000235:genetics*; D024701:Myosin Type V / Q000235:genetics*; D009421:Nervous System Malformations / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D009847:Olivary Nucleus / Q000002:abnormalities* / Q000166:cytology / Q000378:metabolism; D017729:Presynaptic Terminals / Q000378:metabolism / Q000473:pathology; D021381:Protein Transport / Q000502:physiology; D011689:Purkinje Cells / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D011922:Rats, Mutant Strains; D009435:Synaptic Transmission / Q000235:genetics
D003911:Dextrans; C513082:Myo5a protein, mouse; C076397:biotinylated dextran amine; D001710:Biotin; D024701:Myosin Type V; D018995:Myosin Heavy Chains