125I-labeled fasciculin 2: A new tool for quantitation of acetylcholinesterase densities at synaptic sites by EM-autoradiography

Lili Anglister; Jerry Eichler; Maria Szabo; Brigitte Haesaert; Miriam M. Salpeter

doi:10.1016/S0165-0270(98)00015-6

¹²⁵I-labeled fasciculin 2: A new tool for quantitation of acetylcholinesterase densities at synaptic sites by EM-autoradiography

Lili Anglister^*, Jerry Eichler, Maria Szabo, Brigitte Haesaert, Miriam M. Salpeter

^*Corresponding author for this work

Institute for Medical Research Israel-Canada (IMRIC)

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

Radio-iodinated fasciculin 2 (Fas2), a polypeptide anticholinesterase toxin from Mamba venom, was used as a new probe for localizing and quantifying acetylcholinesterase (ACHE) at mouse neuromuscular junctions (NMJs) by quantitative electron microscope autoradiography. We demonstrate that ¹²⁵I-Fas2 binds very specifically to the NMJs of mouse sternomastoid muscles, with very little binding to other regions in the muscles. Junctional AChE-site densities obtained from the autoradiograms were similar to those previously obtained for the same muscles using ³H-DFP. The use of ¹²⁵I- Fas2 with EM-autoradiography is simpler and provides higher resolution and sensitivity, as well as considerably lower non-specific binding than previously attainable with ³H-DFP. The advantages and limitations of this procedure are discussed.

Original language	English
Pages (from-to)	63-71
Number of pages	9
Journal	Journal of Neuroscience Methods
Volume	81
Issue number	1-2
DOIs	https://doi.org/10.1016/S0165-0270(98)00015-6
State	Published - 1 Jun 1998

Bibliographical note

Funding Information:
This research was supported by grants from the Israel Academy of Sciences—Charles H. Revson Foundation 675/94 (to L. Anglister) and from NIH GM 10422 (to M.M. Salpeter). A preliminary account of the potential use of fasciculin for quantitative EM-autoradiography of AChE was presented in Anglister et al., In: Quinn DM et al., editors. Enzymes of the Cholinesterase Family. Plenum, 1995:277–285.

Keywords

Acetylcholinesterase site density
EM-autoradiography
Mouse muscle
Neuromuscular junction
Torpedo G-AChE

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10.1016/S0165-0270(98)00015-6

Cite this

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title = "125I-labeled fasciculin 2: A new tool for quantitation of acetylcholinesterase densities at synaptic sites by EM-autoradiography",

abstract = "Radio-iodinated fasciculin 2 (Fas2), a polypeptide anticholinesterase toxin from Mamba venom, was used as a new probe for localizing and quantifying acetylcholinesterase (ACHE) at mouse neuromuscular junctions (NMJs) by quantitative electron microscope autoradiography. We demonstrate that 125I-Fas2 binds very specifically to the NMJs of mouse sternomastoid muscles, with very little binding to other regions in the muscles. Junctional AChE-site densities obtained from the autoradiograms were similar to those previously obtained for the same muscles using 3H-DFP. The use of 125I- Fas2 with EM-autoradiography is simpler and provides higher resolution and sensitivity, as well as considerably lower non-specific binding than previously attainable with 3H-DFP. The advantages and limitations of this procedure are discussed.",

keywords = "Acetylcholinesterase site density, EM-autoradiography, Mouse muscle, Neuromuscular junction, Torpedo G-AChE",

author = "Lili Anglister and Jerry Eichler and Maria Szabo and Brigitte Haesaert and Salpeter, {Miriam M.}",

note = "Funding Information: This research was supported by grants from the Israel Academy of Sciences—Charles H. Revson Foundation 675/94 (to L. Anglister) and from NIH GM 10422 (to M.M. Salpeter). A preliminary account of the potential use of fasciculin for quantitative EM-autoradiography of AChE was presented in Anglister et al., In: Quinn DM et al., editors. Enzymes of the Cholinesterase Family. Plenum, 1995:277–285.",

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AU - Szabo, Maria

AU - Haesaert, Brigitte

AU - Salpeter, Miriam M.

N1 - Funding Information: This research was supported by grants from the Israel Academy of Sciences—Charles H. Revson Foundation 675/94 (to L. Anglister) and from NIH GM 10422 (to M.M. Salpeter). A preliminary account of the potential use of fasciculin for quantitative EM-autoradiography of AChE was presented in Anglister et al., In: Quinn DM et al., editors. Enzymes of the Cholinesterase Family. Plenum, 1995:277–285.

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N2 - Radio-iodinated fasciculin 2 (Fas2), a polypeptide anticholinesterase toxin from Mamba venom, was used as a new probe for localizing and quantifying acetylcholinesterase (ACHE) at mouse neuromuscular junctions (NMJs) by quantitative electron microscope autoradiography. We demonstrate that 125I-Fas2 binds very specifically to the NMJs of mouse sternomastoid muscles, with very little binding to other regions in the muscles. Junctional AChE-site densities obtained from the autoradiograms were similar to those previously obtained for the same muscles using 3H-DFP. The use of 125I- Fas2 with EM-autoradiography is simpler and provides higher resolution and sensitivity, as well as considerably lower non-specific binding than previously attainable with 3H-DFP. The advantages and limitations of this procedure are discussed.

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