Stimulation-induced modifications in Go proteins examined in giant fused synaptosomes

• Published in: Journal of molecular neuroscience Vol. 20; no. 1; pp. 73 - 80
• Main Authors: Dekel, Noya, Visochek, Leonid, Anis, Yosef, Cohen-Armon, Malka
• Format: Journal Article
• Language: English
• Published: United States Springer Nature B.V 01.02.2003
• Subjects: ADP-Ribosylation Factors - drug effects; ADP-Ribosylation Factors - metabolism; Animals; Antibodies - pharmacology; Brain - metabolism; Cells; Electric Stimulation; GTP-Binding Protein alpha Subunits, Gi-Go; Guanine Nucleotides - pharmacology; Heterotrimeric GTP-Binding Proteins - drug effects; Heterotrimeric GTP-Binding Proteins - metabolism; Liposomes; Male; Patch-Clamp Techniques; Pertussis Toxin - pharmacology; Potassium - pharmacology; Presynaptic Terminals - drug effects; Presynaptic Terminals - metabolism; Rats; Signal Transduction - drug effects; Signal Transduction - physiology; Synaptic Membranes - drug effects; Synaptic Membranes - metabolism; Synaptic Transmission - drug effects; Synaptic Transmission - physiology; Synaptosomes - drug effects; Synaptosomes - metabolism
• ISSN: 0895-8696; 0895-8696; 1559-1166
• DOI: 10.1385/JMN:20:1:73

Synaptoneurosomes (1-3 microm in diameter), prepared from rat brain stem or brain cortex, were fused with liposomes, producing a high yield of giant synaptosomes (10-60 microm in diameter). Single channel currents were measured by using the cell-attach patch-clamp technique. The membrane of the majority of these giant synaptosomes retained the cell membrane selective permeability. However, nonpermeating molecules, such as guanine nucleotides and antibodies directed against GTP-binding region in the alpha-subunit of trimeric GTP-binding proteins, were trapped in the giant synaptosomes during their preparation. Activation of Go proteins was assayed in high [K(+)]-depolarized giant synaptosomes, indicating the advantage of this preparation for tracing signal-transduction mechanisms in stimulated synaptic membranes. Stimulation-induced interactions between membrane proteins, either native or reconstituted, can be studied in the giant synaptosomes.