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Rebuilding a realistic corticostriatal "social network" from dissociated cells.

Garcia-Munoz M, Taillefer E, Pnini R, Vickers C, Miller J, Arbuthnott GW - Front Syst Neurosci (2015)

Bottom Line: The activity of both areas can be recorded in multielectrode arrays or individual patch recordings from pairs of cells.Finally, corticostriatal connections can be severed acutely.This procedure enables determination of the importance of corticostriatal interaction in the resting pattern of activity.

View Article: PubMed Central - PubMed

Affiliation: Brain Mechanisms for Behaviour Unit, Okinawa Institute of Science and Technology Graduate University Okinawa, Japan.

ABSTRACT
Many of the methods available for the study of cortical influences on striatal neurons have serious problems. In vivo the connectivity is so complex that the study of input from an individual cortical neuron to a single striatal cell is nearly impossible. Mixed corticostriatal cultures develop many connections from striatal cells to cortical cells, in striking contrast to the fact that only connections from cortical cells to striatal cells are present in vivo. Furthermore, interneuron populations are over-represented in organotypic cultures. For these reasons, we have developed a method for growing cortical and striatal neurons in separated compartments that allows cortical neurons to innervate striatal cells in culture. The method works equally well for acutely dissociated or cryopreserved neurons and allows a number of manipulations that are not otherwise possible. Either cortical or striatal compartments can be transfected with channel rhodopsins. The activity of both areas can be recorded in multielectrode arrays or individual patch recordings from pairs of cells. Finally, corticostriatal connections can be severed acutely. This procedure enables determination of the importance of corticostriatal interaction in the resting pattern of activity. These cultures also facilitate development of sensitive analytical network methods to track connectivity.

No MeSH data available.


Summary of previous results. Properties of mixed cultures that are also seen in separated cultures include: (A) Synapses are formed between neurons from cortex and striatum. In cultures cortical and striatal cells do not have the obviously different morphologies expected from in vivo investigations. In this false color image, cortical cells are purple, striatal gold, and inhibitory interneurons red. The green dots are glutamatergic boutons stained with antibodies against vGluT1. (B) Spontaneous activity in striatal cells shows “UP” states with action potentials riding on them. (C) Such “UP” states are abolished by application of AMPA receptor antagonist DNQX. (D) Activity of cortical and striatal cells recorded simultaneously are closely related in time (Randall et al., 2011).
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Figure 1: Summary of previous results. Properties of mixed cultures that are also seen in separated cultures include: (A) Synapses are formed between neurons from cortex and striatum. In cultures cortical and striatal cells do not have the obviously different morphologies expected from in vivo investigations. In this false color image, cortical cells are purple, striatal gold, and inhibitory interneurons red. The green dots are glutamatergic boutons stained with antibodies against vGluT1. (B) Spontaneous activity in striatal cells shows “UP” states with action potentials riding on them. (C) Such “UP” states are abolished by application of AMPA receptor antagonist DNQX. (D) Activity of cortical and striatal cells recorded simultaneously are closely related in time (Randall et al., 2011).

Mentions: We have observed that segregated cultures develop synaptic connections and display up-states, spontaneous activity and similar pharmacology to corticostriatal slices. As anticipated because of the presence of cortical cells (Segal et al., 2003), spontaneous depolarizations are blocked by DNQX and APV. When neurons were recorded simultaneously, up states are correlated overall and as reported by Randall et al. (2011) and summarized in Figure 1, it is not clear which neuron is driving the other even in connected pairs.


Rebuilding a realistic corticostriatal "social network" from dissociated cells.

Garcia-Munoz M, Taillefer E, Pnini R, Vickers C, Miller J, Arbuthnott GW - Front Syst Neurosci (2015)

Summary of previous results. Properties of mixed cultures that are also seen in separated cultures include: (A) Synapses are formed between neurons from cortex and striatum. In cultures cortical and striatal cells do not have the obviously different morphologies expected from in vivo investigations. In this false color image, cortical cells are purple, striatal gold, and inhibitory interneurons red. The green dots are glutamatergic boutons stained with antibodies against vGluT1. (B) Spontaneous activity in striatal cells shows “UP” states with action potentials riding on them. (C) Such “UP” states are abolished by application of AMPA receptor antagonist DNQX. (D) Activity of cortical and striatal cells recorded simultaneously are closely related in time (Randall et al., 2011).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403293&req=5

Figure 1: Summary of previous results. Properties of mixed cultures that are also seen in separated cultures include: (A) Synapses are formed between neurons from cortex and striatum. In cultures cortical and striatal cells do not have the obviously different morphologies expected from in vivo investigations. In this false color image, cortical cells are purple, striatal gold, and inhibitory interneurons red. The green dots are glutamatergic boutons stained with antibodies against vGluT1. (B) Spontaneous activity in striatal cells shows “UP” states with action potentials riding on them. (C) Such “UP” states are abolished by application of AMPA receptor antagonist DNQX. (D) Activity of cortical and striatal cells recorded simultaneously are closely related in time (Randall et al., 2011).
Mentions: We have observed that segregated cultures develop synaptic connections and display up-states, spontaneous activity and similar pharmacology to corticostriatal slices. As anticipated because of the presence of cortical cells (Segal et al., 2003), spontaneous depolarizations are blocked by DNQX and APV. When neurons were recorded simultaneously, up states are correlated overall and as reported by Randall et al. (2011) and summarized in Figure 1, it is not clear which neuron is driving the other even in connected pairs.

Bottom Line: The activity of both areas can be recorded in multielectrode arrays or individual patch recordings from pairs of cells.Finally, corticostriatal connections can be severed acutely.This procedure enables determination of the importance of corticostriatal interaction in the resting pattern of activity.

View Article: PubMed Central - PubMed

Affiliation: Brain Mechanisms for Behaviour Unit, Okinawa Institute of Science and Technology Graduate University Okinawa, Japan.

ABSTRACT
Many of the methods available for the study of cortical influences on striatal neurons have serious problems. In vivo the connectivity is so complex that the study of input from an individual cortical neuron to a single striatal cell is nearly impossible. Mixed corticostriatal cultures develop many connections from striatal cells to cortical cells, in striking contrast to the fact that only connections from cortical cells to striatal cells are present in vivo. Furthermore, interneuron populations are over-represented in organotypic cultures. For these reasons, we have developed a method for growing cortical and striatal neurons in separated compartments that allows cortical neurons to innervate striatal cells in culture. The method works equally well for acutely dissociated or cryopreserved neurons and allows a number of manipulations that are not otherwise possible. Either cortical or striatal compartments can be transfected with channel rhodopsins. The activity of both areas can be recorded in multielectrode arrays or individual patch recordings from pairs of cells. Finally, corticostriatal connections can be severed acutely. This procedure enables determination of the importance of corticostriatal interaction in the resting pattern of activity. These cultures also facilitate development of sensitive analytical network methods to track connectivity.

No MeSH data available.