char = C;code = 67 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = -;code = 45 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = g;code = 103 char = a;code = 97 char = n;code = 110 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = d;code = 100 char = i;code = 105 char = s;code = 115 char = p;code = 112 char = l;code = 108 char = a;code = 97 char = y;code = 121 char = ;code = 32 char = u;code = 117 char = n;code = 110 char = i;code = 105 char = q;code = 113 char = u;code = 117 char = e;code = 101 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = i;code = 105 char = t;code = 116 char = y;code = 121 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = c;code = 99 char = h;code = 104 char = e;code = 101 char = m;code = 109 char = i;code = 105 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = i;code = 105 char = m;code = 109 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = c;code = 99 char = u;code = 117 char = l;code = 108 char = t;code = 116 char = u;code = 117 char = r;code = 114 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = h;code = 104 char = i;code = 105 char = p;code = 112 char = p;code = 112 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = m;code = 109 char = p;code = 112 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = n;code = 110 char = e;code = 101 char = u;code = 117 char = r;code = 114 char = o;code = 111 char = n;code = 110 char = s;code = 115 char = .;code = 46 result sql = Calcium-containing organelles display unique reactivity to chemical stimulation in cultured hippocampal neurons.
Calcium-containing organelles display unique reactivity to chemical stimulation in cultured hippocampal neurons.char = C;code = 67 char = u;code = 117 char = l;code = 108 char = t;code = 116 char = u;code = 117 char = r;code = 114 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = r;code = 114 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = h;code = 104 char = i;code = 105 char = p;code = 112 char = p;code = 112 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = m;code = 109 char = p;code = 112 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = n;code = 110 char = e;code = 101 char = u;code = 117 char = r;code = 114 char = o;code = 111 char = n;code = 110 char = s;code = 115 char = ;code = 32 char = g;code = 103 char = r;code = 114 char = o;code = 111 char = w;code = 119 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = g;code = 103 char = l;code = 108 char = a;code = 97 char = s;code = 115 char = s;code = 115 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = v;code = 118 char = e;code = 101 char = r;code = 114 char = s;code = 115 char = l;code = 108 char = i;code = 105 char = p;code = 112 char = s;code = 115 char = ;code = 32 char = f;code = 102 char = o;code = 111 char = r;code = 114 char = ;code = 32 char = 1;code = 49 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = w;code = 119 char = e;code = 101 char = e;code = 101 char = k;code = 107 char = s;code = 115 char = ;code = 32 char = w;code = 119 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = l;code = 108 char = o;code = 111 char = a;code = 97 char = d;code = 100 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = -;code = 45 char = s;code = 115 char = e;code = 101 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = d;code = 100 char = y;code = 121 char = e;code = 101 char = ;code = 32 char = F;code = 70 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = v;code = 118 char = i;code = 105 char = e;code = 101 char = w;code = 119 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = f;code = 102 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = l;code = 108 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = s;code = 115 char = c;code = 99 char = a;code = 97 char = n;code = 110 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = m;code = 109 char = i;code = 105 char = c;code = 99 char = r;code = 114 char = o;code = 111 char = s;code = 115 char = c;code = 99 char = o;code = 111 char = p;code = 112 char = e;code = 101 char = .;code = 46 char = ;code = 32 char = L;code = 76 char = a;code = 97 char = r;code = 114 char = g;code = 103 char = e;code = 101 char = ;code = 32 char = p;code = 112 char = y;code = 121 char = r;code = 114 char = a;code = 97 char = m;code = 109 char = i;code = 105 char = d;code = 100 char = a;code = 97 char = l;code = 108 char = -;code = 45 char = s;code = 115 char = h;code = 104 char = a;code = 97 char = p;code = 112 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = n;code = 110 char = e;code = 101 char = u;code = 117 char = r;code = 114 char = o;code = 111 char = n;code = 110 char = s;code = 115 char = ;code = 32 char = w;code = 119 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = f;code = 102 char = o;code = 111 char = u;code = 117 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = d;code = 100 char = y;code = 121 char = e;code = 101 char = -;code = 45 char = a;code = 97 char = c;code = 99 char = c;code = 99 char = u;code = 117 char = m;code = 109 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = g;code = 103 char = a;code = 97 char = n;code = 110 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = i;code = 105 char = r;code = 114 char = ;code = 32 char = s;code = 115 char = o;code = 111 char = m;code = 109 char = a;code = 97 char = t;code = 116 char = a;code = 97 char = ,;code = 44 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = i;code = 105 char = m;code = 109 char = a;code = 97 char = r;code = 114 char = i;code = 105 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = a;code = 97 char = r;code = 114 char = o;code = 111 char = u;code = 117 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = n;code = 110 char = u;code = 117 char = c;code = 99 char = l;code = 108 char = e;code = 101 char = i;code = 105 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = n;code = 110 char = e;code = 101 char = a;code = 97 char = r;code = 114 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = b;code = 98 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = i;code = 105 char = r;code = 114 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = i;code = 105 char = m;code = 109 char = a;code = 97 char = r;code = 114 char = y;code = 121 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = n;code = 110 char = d;code = 100 char = r;code = 114 char = i;code = 105 char = t;code = 116 char = e;code = 101 char = s;code = 115 char = .;code = 46 char = ;code = 32 char = T;code = 84 char = h;code = 104 char = e;code = 101 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = g;code = 103 char = a;code = 97 char = n;code = 110 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = v;code = 118 char = a;code = 97 char = r;code = 114 char = i;code = 105 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = s;code = 115 char = i;code = 105 char = z;code = 122 char = e;code = 101 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = c;code = 99 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = y;code = 121 char = ;code = 32 char = o;code = 111 char = v;code = 118 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = w;code = 119 char = e;code = 101 char = e;code = 101 char = k;code = 107 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = c;code = 99 char = u;code = 117 char = l;code = 108 char = t;code = 116 char = u;code = 117 char = r;code = 114 char = e;code = 101 char = ,;code = 44 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = w;code = 119 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = n;code = 110 char = o;code = 111 char = t;code = 116 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = e;code = 101 char = n;code = 110 char = d;code = 100 char = o;code = 111 char = p;code = 112 char = l;code = 108 char = a;code = 97 char = s;code = 115 char = m;code = 109 char = i;code = 105 char = c;code = 99 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = t;code = 116 char = i;code = 105 char = c;code = 99 char = u;code = 117 char = l;code = 108 char = u;code = 117 char = m;code = 109 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = m;code = 109 char = i;code = 105 char = t;code = 116 char = o;code = 111 char = c;code = 99 char = h;code = 104 char = o;code = 111 char = n;code = 110 char = d;code = 100 char = r;code = 114 char = i;code = 105 char = a;code = 97 char = .;code = 46 char = ;code = 32 char = T;code = 84 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = F;code = 70 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = c;code = 99 char = e;code = 101 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = -;code = 45 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = g;code = 103 char = a;code = 97 char = n;code = 110 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = (;code = 40 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = );code = 41 char = ;code = 32 char = w;code = 119 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = t;code = 116 char = r;code = 114 char = a;code = 97 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = q;code = 113 char = u;code = 117 char = e;code = 101 char = n;code = 110 char = c;code = 99 char = h;code = 104 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = b;code = 98 char = y;code = 121 char = ;code = 32 char = e;code = 101 char = x;code = 120 char = p;code = 112 char = o;code = 111 char = s;code = 115 char = u;code = 117 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = M;code = 77 char = n;code = 110 char = 2;code = 50 char = +;code = 43 char = ,;code = 44 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = d;code = 100 char = i;code = 105 char = c;code = 99 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = d;code = 100 char = y;code = 121 char = e;code = 101 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = g;code = 103 char = e;code = 101 char = n;code = 110 char = u;code = 117 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = 2;code = 50 char = +;code = 43 char = ];code = 93 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = p;code = 112 char = o;code = 111 char = r;code = 114 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = n;code = 110 char = o;code = 111 char = t;code = 116 char = ;code = 32 char = j;code = 106 char = u;code = 117 char = s;code = 115 char = t;code = 116 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = a;code = 97 char = c;code = 99 char = c;code = 99 char = u;code = 117 char = m;code = 109 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = F;code = 70 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = d;code = 100 char = y;code = 121 char = e;code = 101 char = .;code = 46 char = ;code = 32 char = R;code = 82 char = e;code = 101 char = c;code = 99 char = o;code = 111 char = v;code = 118 char = e;code = 101 char = r;code = 114 char = y;code = 121 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = c;code = 99 char = e;code = 101 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = f;code = 102 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = w;code = 119 char = a;code = 97 char = s;code = 115 char = h;code = 104 char = o;code = 111 char = u;code = 117 char = t;code = 116 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = M;code = 77 char = n;code = 110 char = 2;code = 50 char = +;code = 43 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = v;code = 118 char = o;code = 111 char = l;code = 108 char = v;code = 118 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = p;code = 112 char = s;code = 115 char = i;code = 105 char = g;code = 103 char = a;code = 97 char = r;code = 114 char = g;code = 103 char = i;code = 105 char = n;code = 110 char = -;code = 45 char = s;code = 115 char = e;code = 101 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = c;code = 99 char = e;code = 101 char = s;code = 115 char = s;code = 115 char = .;code = 46 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = p;code = 112 char = o;code = 111 char = n;code = 110 char = d;code = 100 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = i;code = 105 char = m;code = 109 char = u;code = 117 char = l;code = 108 char = i;code = 105 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = e;code = 101 char = v;code = 118 char = o;code = 111 char = k;code = 107 char = e;code = 101 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = r;code = 114 char = i;code = 105 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = y;code = 121 char = t;code = 116 char = o;code = 111 char = s;code = 115 char = o;code = 111 char = l;code = 108 char = i;code = 105 char = c;code = 99 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = 2;code = 50 char = +;code = 43 char = ];code = 93 char = ;code = 32 char = (;code = 40 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = i;code = 105 char = );code = 41 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = u;code = 117 char = n;code = 110 char = i;code = 105 char = q;code = 113 char = u;code = 117 char = e;code = 101 char = ;code = 32 char = m;code = 109 char = a;code = 97 char = n;code = 110 char = n;code = 110 char = e;code = 101 char = r;code = 114 char = ;;code = 59 char = ;code = 32 char = p;code = 112 char = e;code = 101 char = r;code = 114 char = f;code = 102 char = u;code = 117 char = s;code = 115 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = l;code = 108 char = o;code = 111 char = n;code = 110 char = g;code = 103 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = r;code = 114 char = i;code = 105 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = (;code = 40 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = C;code = 67 char = );code = 41 char = ,;code = 44 char = ;code = 32 char = w;code = 119 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = t;code = 116 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = t;code = 116 char = r;code = 114 char = a;code = 97 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = r;code = 114 char = i;code = 105 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = i;code = 105 char = .;code = 46 char = ;code = 32 char = P;code = 80 char = u;code = 117 char = l;code = 108 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = a;code = 97 char = p;code = 112 char = p;code = 112 char = l;code = 108 char = i;code = 105 char = c;code = 99 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = a;code = 97 char = l;code = 108 char = s;code = 115 char = o;code = 111 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = f;code = 102 char = a;code = 97 char = s;code = 115 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = e;code = 101 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = C;code = 67 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = i;code = 105 char = .;code = 46 char = ;code = 32 char = G;code = 71 char = l;code = 108 char = u;code = 117 char = t;code = 116 char = a;code = 97 char = m;code = 109 char = a;code = 97 char = t;code = 116 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = t;code = 116 char = r;code = 114 char = a;code = 97 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = r;code = 114 char = i;code = 105 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = b;code = 98 char = o;code = 111 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = i;code = 105 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = C;code = 67 char = ,;code = 44 char = ;code = 32 char = f;code = 102 char = o;code = 111 char = l;code = 108 char = l;code = 108 char = o;code = 111 char = w;code = 119 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = b;code = 98 char = y;code = 121 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = l;code = 108 char = o;code = 111 char = n;code = 110 char = g;code = 103 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = f;code = 102 char = a;code = 97 char = l;code = 108 char = l;code = 108 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = C;code = 67 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = b;code = 98 char = e;code = 101 char = l;code = 108 char = o;code = 111 char = w;code = 119 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = t;code = 116 char = ;code = 32 char = l;code = 108 char = e;code = 101 char = v;code = 118 char = e;code = 101 char = l;code = 108 char = .;code = 46 char = ;code = 32 char = T;code = 84 char = h;code = 104 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = f;code = 102 char = a;code = 97 char = l;code = 108 char = l;code = 108 char = ;code = 32 char = w;code = 119 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = b;code = 98 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = k;code = 107 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = b;code = 98 char = y;code = 121 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = c;code = 99 char = u;code = 117 char = b;code = 98 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = p;code = 112 char = s;code = 115 char = i;code = 105 char = g;code = 103 char = a;code = 97 char = r;code = 114 char = g;code = 103 char = i;code = 105 char = n;code = 110 char = .;code = 46 char = ;code = 32 char = R;code = 82 char = y;code = 121 char = a;code = 97 char = n;code = 110 char = o;code = 111 char = d;code = 100 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = b;code = 98 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = k;code = 107 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = y;code = 121 char = t;code = 116 char = o;code = 111 char = s;code = 115 char = o;code = 111 char = l;code = 108 char = i;code = 105 char = c;code = 99 char = ;code = 32 char = e;code = 101 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = s;code = 115 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = b;code = 98 char = u;code = 117 char = t;code = 116 char = ;code = 32 char = n;code = 110 char = o;code = 111 char = t;code = 116 char = ;code = 32 char = i;code = 105 char = t;code = 116 char = s;code = 115 char = ;code = 32 char = e;code = 101 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = ];code = 93 char = C;code = 67 char = .;code = 46 char = ;code = 32 char = A;code = 65 char = ;code = 32 char = c;code = 99 char = l;code = 108 char = e;code = 101 char = a;code = 97 char = r;code = 114 char = ;code = 32 char = d;code = 100 char = i;code = 105 char = s;code = 115 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = b;code = 98 char = e;code = 101 char = t;code = 116 char = w;code = 119 char = e;code = 101 char = e;code = 101 char = n;code = 110 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = k;code = 107 char = n;code = 110 char = o;code = 111 char = w;code = 119 char = n;code = 110 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = w;code = 119 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = s;code = 115 char = e;code = 101 char = e;code = 101 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = d;code = 100 char = i;code = 105 char = g;code = 103 char = i;code = 105 char = t;code = 116 char = o;code = 111 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = -;code = 45 char = p;code = 112 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = e;code = 101 char = a;code = 97 char = b;code = 98 char = i;code = 105 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = ;;code = 59 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = s;code = 115 char = e;code = 101 char = ,;code = 44 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = m;code = 109 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = F;code = 70 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = p;code = 112 char = o;code = 111 char = r;code = 114 char = t;code = 116 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = c;code = 99 char = h;code = 104 char = a;code = 97 char = n;code = 110 char = g;code = 103 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = ,;code = 44 char = ;code = 32 char = i;code = 105 char = .;code = 46 char = e;code = 101 char = .;code = 46 char = ,;code = 44 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = d;code = 100 char = u;code = 117 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = F;code = 70 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = -;code = 45 char = 3;code = 51 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = c;code = 99 char = e;code = 101 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = p;code = 112 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = e;code = 101 char = a;code = 97 char = b;code = 98 char = i;code = 105 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = ,;code = 44 char = ;code = 32 char = w;code = 119 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = t;code = 116 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = i;code = 105 char = l;code = 108 char = l;code = 108 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = c;code = 99 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = [;code = 91 char = C;code = 67 char = a;code = 97 char = ];code = 93 char = C;code = 67 char = .;code = 46 char = ;code = 32 char = A;code = 65 char = ;code = 32 char = p;code = 112 char = o;code = 111 char = s;code = 115 char = s;code = 115 char = i;code = 105 char = b;code = 98 char = l;code = 108 char = e;code = 101 char = ;code = 32 char = r;code = 114 char = o;code = 111 char = l;code = 108 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = g;code = 103 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = l;code = 108 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = ;code = 32 char = f;code = 102 char = r;code = 114 char = o;code = 111 char = m;code = 109 char = ;code = 32 char = r;code = 114 char = y;code = 121 char = a;code = 97 char = n;code = 110 char = o;code = 111 char = d;code = 100 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = -;code = 45 char = s;code = 115 char = e;code = 101 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = w;code = 119 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = d;code = 100 char = i;code = 105 char = c;code = 99 char = a;code = 97 char = t;code = 116 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = b;code = 98 char = y;code = 121 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = b;code = 98 char = s;code = 115 char = e;code = 101 char = r;code = 114 char = v;code = 118 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = -;code = 45 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = ;code = 32 char = e;code = 101 char = x;code = 120 char = h;code = 104 char = i;code = 105 char = b;code = 98 char = i;code = 105 char = t;code = 116 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = l;code = 108 char = a;code = 97 char = r;code = 114 char = g;code = 103 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = f;code = 102 char = a;code = 97 char = s;code = 115 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = p;code = 112 char = o;code = 111 char = n;code = 110 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = d;code = 100 char = i;code = 105 char = d;code = 100 char = ;code = 32 char = n;code = 110 char = o;code = 111 char = t;code = 116 char = ;code = 32 char = h;code = 104 char = a;code = 97 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = m;code = 109 char = .;code = 46 char = ;code = 32 char = W;code = 87 char = e;code = 101 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = p;code = 112 char = o;code = 111 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = a;code = 97 char = t;code = 116 char = ;code = 32 char = C;code = 67 char = C;code = 67 char = O;code = 79 char = s;code = 115 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = n;code = 110 char = s;code = 115 char = t;code = 116 char = i;code = 105 char = t;code = 116 char = u;code = 117 char = t;code = 116 char = e;code = 101 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = u;code = 117 char = n;code = 110 char = i;code = 105 char = q;code = 113 char = u;code = 117 char = e;code = 101 char = ;code = 32 char = f;code = 102 char = u;code = 117 char = n;code = 110 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = m;code = 109 char = p;code = 112 char = a;code = 97 char = r;code = 114 char = t;code = 116 char = m;code = 109 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = v;code = 118 char = o;code = 111 char = l;code = 108 char = v;code = 118 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = l;code = 108 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = ;code = 32 char = f;code = 102 char = r;code = 114 char = o;code = 111 char = m;code = 109 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = c;code = 99 char = i;code = 105 char = u;code = 117 char = m;code = 109 char = -;code = 45 char = s;code = 115 char = e;code = 101 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = .;code = 46 char = <;code = 60 char = b;code = 98 char = r;code = 114 char = /;code = 47 char = >;code = 62 result sql = Cultured rat hippocampal neurons grown on glass coverslips for 1-3 weeks were loaded with the calcium-sensitive fluorescent dye Fluo-3 and viewed with a confocal laser scanning microscope. Large pyramidal-shaped neurons were found to contain dye-accumulating organelles in their somata, primarily around nuclei and near the base of their primary dendrites. These organelles varied in size and increased in density over weeks in culture, and were not colocalized with the endoplasmic reticulum or with mitochondria. The Fluo-3 fluorescence in these calcium-containing organelles (CCOs) was transiently quenched by exposure to Mn2+, indicating that the dye is a genuine [Ca2+] reporter and is not just a site of accumulating Fluo-3 dye. Recovery of fluorescence in the CCOs after washout of Mn2+ involved activation of a thapsigargin-sensitive process. CCOs responded to stimuli that evoke a rise of cytosolic [Ca2+] ([Ca]i) in a unique manner; perfusion of caffeine caused a prolonged rise of [Ca] in the CCOs ([Ca]C), whereas it caused only a transient rise of [Ca]i. Pulse application of caffeine also caused a faster effect on [Ca]C than on [Ca]i. Glutamate caused a transient rise of both [Ca]i and [Ca]C, followed by a prolonged fall of only [Ca]C to below rest level. This fall was blocked by preincubation with thapsigargin. Ryanodine blocked the cytosolic effects of caffeine but not its effect on [C]C. A clear distinction between CCOs and the known calcium stores was seen in digitonin-permeabilized cells; in these, remaining Fluo-3 reported changes in store calcium, i.e., caffeine caused a reduction in Fluo-3 fluorescence in permeabilized cells, whereas it still caused an increase in [Ca]C. A possible role of CCOs in regulation of release of calcium from ryanodine-sensitive stores was indicated by the observation that CCO-containing cells exhibited a larger and faster response to caffeine than cells that did not have them. We propose that CCOs constitute a unique functional compartment involved in release of calcium from calcium-sensitive stores.
Publication: J. Neurosci. 1997 Mar 1 ;17(5)::1670-82.
Publication Date: 1997 Mar 1
Study Author(s): Korkotian, E;Segal, M;
Institution: Department of Neurobiology, The Weizmann Institute, Rehovot 76100, Israel.
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Cultured rat hippocampal neurons grown on glass coverslips for 1-3 weeks were loaded with the calcium-sensitive fluorescent dye Fluo-3 and viewed with a confocal laser scanning microscope. Large pyramidal-shaped neurons were found to contain dye-accumulating organelles in their somata, primarily around nuclei and near the base of their primary dendrites. These organelles varied in size and increased in density over weeks in culture, and were not colocalized with the endoplasmic reticulum or with mitochondria. The Fluo-3 fluorescence in these calcium-containing organelles (CCOs) was transiently quenched by exposure to Mn2+, indicating that the dye is a genuine [Ca2+] reporter and is not just a site of accumulating Fluo-3 dye. Recovery of fluorescence in the CCOs after washout of Mn2+ involved activation of a thapsigargin-sensitive process. CCOs responded to stimuli that evoke a rise of cytosolic [Ca2+] ([Ca]i) in a unique manner; perfusion of caffeine caused a prolonged rise of [Ca] in the CCOs ([Ca]C), whereas it caused only a transient rise of [Ca]i. Pulse application of caffeine also caused a faster effect on [Ca]C than on [Ca]i. Glutamate caused a transient rise of both [Ca]i and [Ca]C, followed by a prolonged fall of only [Ca]C to below rest level. This fall was blocked by preincubation with thapsigargin. Ryanodine blocked the cytosolic effects of caffeine but not its effect on [C]C. A clear distinction between CCOs and the known calcium stores was seen in digitonin-permeabilized cells; in these, remaining Fluo-3 reported changes in store calcium, i.e., caffeine caused a reduction in Fluo-3 fluorescence in permeabilized cells, whereas it still caused an increase in [Ca]C. A possible role of CCOs in regulation of release of calcium from ryanodine-sensitive stores was indicated by the observation that CCO-containing cells exhibited a larger and faster response to caffeine than cells that did not have them. We propose that CCOs constitute a unique functional compartment involved in release of calcium from calcium-sensitive stores.
PMID: 9030626