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Bi / CNS 150 Lecture 8 Synaptic inhibition; cable properties of neurons; electrical integration in cerebellum Wednesday, October 15, 2013 Henry

Lester

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Nicotinic ACh, GABAA , and glycine receptors look alike at this resolution (prev. lecture) ~ 2200 amino acids in 5 chains (“subunits”), MW ~ 2.5 x 106 Binding g region

Membrane region

Colored by secondary structure

Colored by subunit (chain)

Cytosolic region

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The pentameric GABAA and glycine receptors look like ACh receptors; but they are permeable to anions (mostly Cl-, of course)
1. -amino-butyric acid (GABA) is the principal inhibitory transmitter in the brain. 2 Glycine is the dominant inhibitory transmitter in the spinal cord & hindbrain 2. hindbrain. GABAA receptors are more variable than glycine receptors in subunit composition and therefore in kinetic behavior.

. . Cation channels become anion channels with only one amino acid change per subunit, subunit in this approximate location

Like a previous lecture

current flows outward Negative to Erev.A Synapse “pushes” the Membrane Potential Toward the Reversal Potential (Erev) for the synaptic Channels ACh and glutamate receptors flux Na+ and K+. current flows inward ENa +60 60 +40 +20 20 -5 -20 Resting -50 potential -80 EK Like Figure 10-11 -100 4 . Positive to Erev. the current through g open receptors is zero. M b Membrane potential t ti l +80 At GABAA and glycine receptors. Erev is near ECl ~ -70 mV At Erev . and Erev ~ 0 mV. (and in some cases Ca2+).

Lunesta are derivatives) The natural ligand binds at subunit interfeces (like ACh at ACh receptors) phenobarbital site is unceratin 5 . (Ambien.Pharmacology of GABAA Receptors: activators Benzodiazepines (= BZ below): Valium (diazepam).

GABA and glycine also make cation- interactions Y198 C2 W149 B Y93 A Y190 C1 non-W55 D (Muscle Nicotinic numbering) 6 . . lecture) .The AChBP interfacial “aromatic box” occupied by nicotine (prev. .

GABAA and Glycine blockers bind either at the agonist site or in the channel Bicuculline (GABAA receptor ( p Agonist site it Strychnine (glycine receptor) t ) Picrotoxin (GABAA & glycine receptors) .

How does the receptor transduce binding into channel gating? (prev. . . Both ideas are also in play for GABA or glycine receptors S i l? Swivel? Miyazawa. Nature 2003 CLOSED Twist? Corringer. lecture) . J Physiol 2010 OPEN 8 .

GABA. dopamine dopamine.We have Completed our Survey of Synaptic Receptors A. tyrosine) receptor-channels Most ^ Figure 10-7 9 . (invert GluCl (invert. Serotonin 5-HT3. ACh. GluCl.

Parts of two generalized CNS neurons Presynaptic neuron Excitatory Inhibitory terminal terminal axon presynaptic terminal Postsynaptic neuron node of Ranvier initial myelin segment apical dendrites (apex) little hill axon hillock hill k cell (base) basal body dendrites (soma) presynaptic nucleus terminal synaptic cleft direction of information flow postsynaptic dendrite Like Figure 2-1 (rotated) 10 .

it exemplifies prepre and postsynaptic structures structures.The cerebellum: a famous circuit in neuroscience. Molecular layer Purkinje cell layer Ganule cell layer White matter 10% of the neurons in the CNS are cerebellar granule cells Figure 42-4 11 . In today’s lecture.

A plurality of synapses in the CNS (> 1013 ) occur between parallel fiber axons and Purkinje cell dendritic spines Molecular layer y 500 nm 12 .

Types of synapses (Don’t mind the Type I. Type II stuff) Figure 10-3 13 .

Types of synaptic integration 1. Capacitive filtering 2 Spatial 2. Temporal e po a A. 3. Excitatory-inhibitory 14 . Molecular lifetimes B.

Molecular lifetimes Concentration of high acetylcholine at NMJ (because of 0 acetylcholinesterase.Previous lecture Synaptic integration 1A. turnover time ~ 100 μs) State 1 closed k21 State 2 open all molecules begin here at t= 0 units: s-1 Number of open p channels ms 15 .

acetylcholinesterase is present at densities of > 1000 / μm2 near each synapse synapse.At the nerve-muscle synapse. 16 . probably high enough to sequester each transmitter molecule as it leaves a receptor (more in a few slides). high enough to destroy each transmitter molecule as it leaves a receptor What causes the ~ δ-function of glutamate & GABA at CNS synapses? Na+ -coupled transporters for glutamate & GABA are present at densities of > 1000 / μm2 near each synapse.

V  C  IC Figure 9-6 17 . Capacitive filtering I C  C dV dT .Synaptic Integration 1B.

1B. V  C  IC Improved from Figure 10-14 .33 mm) ~ 100 pA 2 mV 25 ms 18 I C  C dV dT . Temporal Summation Recording 2. Spatial summation Recording Axon Axon Synaptic y p Current Synaptic Potential Long time constant (100 ms) Vm Short time constant Vm (20 ms) Synaptic Current Synaptic S ti Potential Long g length g constant (1 mm) Short length constant (0.

19 . . If dendrites were passive. .1. Neurobiol 64:75. Excitatory synapses V EPSP measured in soma V EPSP measured in dendrite Gulledge & Stuart (2005) J. they would act like leaky cables .

colocalized EPSPs (two individual trials) Nearly simultaneous.edu/neuron/static/about/stylmn.neuron. colocalized EPSP EPSPs (two individual trials) Simultaneous. .html Gulledge & Stuart (2005) J. . and passively integrate inputs . . simulation and run the movie movie.yale. Neurobiol 64:75. Spatially p y distinct EPSPs Inspect the simulation.. V ∆t = 0 V ∆t = 5 ms V ∆t = 0 Prolonged rising phase Simultaneous. 20 . . at http://www.

.. . . . 21 . Gulledge & Stuart (2005) J. Neurobiol 64:75. but two-photon microscopes allow researchers to visualize patch-clamped dendrites in living animals .

. 2001 22 .. Brain Res. dendrites are not passive. J Physiol (1995) Whitaker. They have Na channels Now break the patch. . patch to fill the cell with dye: immunocytochemistry Averaged traces * = axon hillock 25 μm Magee & Johnston.

brain slice . voltage-gated lt t d Na N + and dC Ca2+ channels h l in dendrites lead to partial “backpropagation” of p . 23 . action potentials. . implying that parts of cells can process signals semi-independently. . Neurobiol 64:75. Stay tuned! Gulledge & Stuart (2005) J.

3. Excitatory-inhibitory integration: The “veto principle” of inhibitory transmission Inhibitory synapses work best when they are “near“ near the excitatory event they will inhibit. Inhibitory synapses on cell bodies and initial segments do a good job of inhibiting spikes 24 . inhibit “Near” means < one cable length. A. Inhibitory synapses on dendrites do a good job of inhibiting EPSPs on nearby spines B.

“Veto” inhibition at the axon initial segment: Schematic of a GABAergic “chandelier chandelier cell” cell in human cerebral cortex Inhibitory Ch d li Chandelier Cell Ch terminals Ch. 1807 25 . axon Pyramidal Cells Ch terminals from Felipe et al. Brain (1999) 122.

Now we localize the inhibitory “vetos” of cerebellar Purkinje cells by “pinceaux” pinceaux (paintbrushes) of basket cells Molecular layer Purkinje cell layer Ganule cell layer White matter Figure 42-4 26 .

How to localize and quantify inhibitory synapses NH2 A fusion protein: GABA transporter (GAT1)-GFP 27 .

cerebellum 28 .

<Immunocytochemistry For GABA transporter Molecular layer (basket cells stain) Purkinje cell layer “pinceux” (paintbrushes) stain heavily Granule cell layer 29 .

showing soma-hillock “veto” Granule cell layer 30 .<Immunocytochemistry For GABA transporter mGAT1 GFP knock-in fluorescence > Molecular layer (basket cells stain) Purkinje cell layer “pinceaux” stain heavily.

GAT1-GFP expression in cerebellum: basket cell terminals in molecular layer. Showing dendritic “veto” veto GABA transporter density is ~1000/(μm2) 50 m 31 .

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