Yoav Livneh

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Presentation transcript:

Yoav Livneh

neurogenesis and migration Refinement of synaptic connections outgrowth of axons and dendrites, and synaptogenesis Neuronal development (generally)

Adult neurogenesis (hippocampus) * also occurs in the adult olfactory bulb

Neuronal development (in the adult hippocampus) Ge et al., J Physiol, 2008

GABA in neuronal development Ben-Ari, Nature Reviews: Neuroscience, 2002

1. Genesis of neurons (and migration). 2. Outgrowth of axons and dendrites, and synaptogenesis. 3. Refinement of synaptic connections.

Development of GABA (and glutamate) currents in adult-born hippocampal neurons tonic GABA currents evoked GABA currents evoked Glu. currents 3 dpi7 dpi14 dpi28 dpi DCX (doublecortin) - a marker for immature neurons Bicuculine – GABAa-R antagonist CNQX – AMPA-R antagonist

The big question – What is the role of these tonic excitatory GABA currents?

Exploration and manipulation of GABA currents in adult-born hippocampal neurons exc.inh. - GABA is excitatory from 3 to ~14 dpi. - Afterwards GABA currents mature to become inhibitory. - shRNA-mediated knockdown of NKCC1 (chloride transporter) eliminated the excitatory nature of GABA (high [Cl] in is not maintained). ~inh. Perforated patch – initially keeps the original intracellular content. Whole-cell patch – content is very rapidly replaced with the electrode’s content.

How will this manipulation affect adult-born neuron development?

Effects of eliminating excitatory GABA currents – part I (dendritic development) Excitatory GABA currents are essential (necessary) for proper dendritic development - Shorter total dendritic length. - Less branches. - Occupy a smaller volume of space.

Effects of enhancing excitatory GABA currents The opposite, complementary, trends were observed with administration of a GABA agonist (suggesting it might also be sufficient). (should be positively correlated) Pentobarbital – GABA agonist

Effects of eliminating excitatory GABA currents – part II (inhibitory synaptic integration) Excitatory GABA currents are essential for proper inhibitory synaptic integration evokedspontaneous - Less neurons with evoked inhibitory responses. - Lower evoked response amplitudes. - Lower spontaneous response frequencies (but similar amplitudes).

Effects of eliminating excitatory GABA currents – part II (excitatory synaptic integration) Excitatory GABA currents are essential for proper excitatory synaptic integration evoked spontaneous - Dramatically less neurons with evoked excitatory responses. - Lower evoked response amplitudes. - Lower spontaneous response frequencies (but similar amplitudes).

Effects of eliminating excitatory GABA currents from birth (KO mice) The same trends were observed with NKCC1 KO mice.

Conclusions and interpretations Conclusions: -At the initial stages of the development of adult-born neurons, GABA is excitatory due to high intracellular Cl concentration (mediated by the transporter NKCC1). - This excitatory action of GABA is essential (necessary and perhaps sufficient) for proper development of dendrites and synaptic connections. Interpretations: - New neurons initially “listen” to network activity, before they “talk” (what they “hear” might affect their integration). - Tonic excitatory GABA might act through low-threshold (-57 mV) T- type calcium channels (expressed in these neurons). Calcium then initiates changes in gene expression, cytoskeletal rearrangement, etc.

Questions?