Ca2+ dynamics in synaptosomes isolated from the squid optic lobe

Juan C  Benech; Pedro A  Lima; José R  Sotelo; Euan R Brown

doi:10.1002/1097-4547(20001215)62:6<840::AID-JNR11>3.0.CO;2-8

Ca2+ dynamics in synaptosomes isolated from the squid optic lobe

Juan C Benech, Pedro A Lima, José R Sotelo, Euan R Brown

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8 Citations (Scopus)

Abstract

Synaptosomes from the optic lobes of squid (Loligo forbesi) were prepared by homogenization and allowed to settle onto glass coverslips. Synaptosomes were loaded with Ca2+ sensitive dyes (Fura-2 AM, Calcium Green-1 AM and Calcium Green-5N AM), visualized by light microscopy and Ca2+ sensitive fluorescence signals recorded and analyzed. With Fura-2, resting Ca2+ was found to be 80 nM (n = 10, SEM 5.7). Addition of K+ (30 mM), caffeine (3 mM) and thapsigargin (10 muM) evoked transient increases in cytoplasmic Ca2+. Addition of BAPTA-AM (20 muM) decreased intrasynaptosomal free Ca2+. Similar results were obtained with Calcium Green-1 AM but not with Calcium Green-5N AM. We conclude that synaptosomes from the squid optic lobe posses intact membranes and mechanisms to regulate intrasynaptosomal free [Ca2+], as well as caffeine sensitive Ca2+ stores. The results of this study are discussed with respect to the role of Ca2+ in presynaptic protein synthesis. (C) 2000 Wiley-Liss, Inc.

Original language	English
Pages (from-to)	840-846
Number of pages	7
Journal	Journal of Neuroscience Research
Volume	62
Issue number	6
DOIs	https://doi.org/10.1002/1097-4547(20001215)62:6<840::AID-JNR11>3.0.CO;2-8
Publication status	Published - 15 Dec 2000

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title = "Ca2+ dynamics in synaptosomes isolated from the squid optic lobe",

abstract = "Synaptosomes from the optic lobes of squid (Loligo forbesi) were prepared by homogenization and allowed to settle onto glass coverslips. Synaptosomes were loaded with Ca2+ sensitive dyes (Fura-2 AM, Calcium Green-1 AM and Calcium Green-5N AM), visualized by light microscopy and Ca2+ sensitive fluorescence signals recorded and analyzed. With Fura-2, resting Ca2+ was found to be 80 nM (n = 10, SEM 5.7). Addition of K+ (30 mM), caffeine (3 mM) and thapsigargin (10 muM) evoked transient increases in cytoplasmic Ca2+. Addition of BAPTA-AM (20 muM) decreased intrasynaptosomal free Ca2+. Similar results were obtained with Calcium Green-1 AM but not with Calcium Green-5N AM. We conclude that synaptosomes from the squid optic lobe posses intact membranes and mechanisms to regulate intrasynaptosomal free [Ca2+], as well as caffeine sensitive Ca2+ stores. The results of this study are discussed with respect to the role of Ca2+ in presynaptic protein synthesis. (C) 2000 Wiley-Liss, Inc.",

author = "Benech, {Juan C} and Lima, {Pedro A} and Sotelo, {Jos{\'e} R} and Brown, {Euan R}",

year = "2000",

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doi = "10.1002/1097-4547(20001215)62:6<840::AID-JNR11>3.0.CO;2-8",

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TY - JOUR

T1 - Ca2+ dynamics in synaptosomes isolated from the squid optic lobe

AU - Benech, Juan C

AU - Lima, Pedro A

AU - Sotelo, José R

AU - Brown, Euan R

PY - 2000/12/15

Y1 - 2000/12/15

N2 - Synaptosomes from the optic lobes of squid (Loligo forbesi) were prepared by homogenization and allowed to settle onto glass coverslips. Synaptosomes were loaded with Ca2+ sensitive dyes (Fura-2 AM, Calcium Green-1 AM and Calcium Green-5N AM), visualized by light microscopy and Ca2+ sensitive fluorescence signals recorded and analyzed. With Fura-2, resting Ca2+ was found to be 80 nM (n = 10, SEM 5.7). Addition of K+ (30 mM), caffeine (3 mM) and thapsigargin (10 muM) evoked transient increases in cytoplasmic Ca2+. Addition of BAPTA-AM (20 muM) decreased intrasynaptosomal free Ca2+. Similar results were obtained with Calcium Green-1 AM but not with Calcium Green-5N AM. We conclude that synaptosomes from the squid optic lobe posses intact membranes and mechanisms to regulate intrasynaptosomal free [Ca2+], as well as caffeine sensitive Ca2+ stores. The results of this study are discussed with respect to the role of Ca2+ in presynaptic protein synthesis. (C) 2000 Wiley-Liss, Inc.

AB - Synaptosomes from the optic lobes of squid (Loligo forbesi) were prepared by homogenization and allowed to settle onto glass coverslips. Synaptosomes were loaded with Ca2+ sensitive dyes (Fura-2 AM, Calcium Green-1 AM and Calcium Green-5N AM), visualized by light microscopy and Ca2+ sensitive fluorescence signals recorded and analyzed. With Fura-2, resting Ca2+ was found to be 80 nM (n = 10, SEM 5.7). Addition of K+ (30 mM), caffeine (3 mM) and thapsigargin (10 muM) evoked transient increases in cytoplasmic Ca2+. Addition of BAPTA-AM (20 muM) decreased intrasynaptosomal free Ca2+. Similar results were obtained with Calcium Green-1 AM but not with Calcium Green-5N AM. We conclude that synaptosomes from the squid optic lobe posses intact membranes and mechanisms to regulate intrasynaptosomal free [Ca2+], as well as caffeine sensitive Ca2+ stores. The results of this study are discussed with respect to the role of Ca2+ in presynaptic protein synthesis. (C) 2000 Wiley-Liss, Inc.

U2 - 10.1002/1097-4547(20001215)62:6<840::AID-JNR11>3.0.CO;2-8

DO - 10.1002/1097-4547(20001215)62:6<840::AID-JNR11>3.0.CO;2-8

M3 - Article

SN - 0360-4012

VL - 62

SP - 840

EP - 846

JO - Journal of Neuroscience Research

JF - Journal of Neuroscience Research

IS - 6

ER -

Ca2+ dynamics in synaptosomes isolated from the squid optic lobe

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