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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Evidence for enhancement of gap junctional coupling between rat island of Calleja granule cells in vitro by the activation of dopamine D3 receptors.

1. Using patch-clamp techniques, we have studied actions of dopamine and related compounds on granule neurones within the islands of Calleja in vitro, in slices if approximately 200 microns thickness or as groups of varying cell number following enzymic digestion. 2. Prior to agonist application, island of Calleja granule cells displayed spontaneous stepwise shifts in whole-cell conductance ranging from 104 to 632 pS. The reversal potentials of these conductance changes ranged widely and matched the distribution of the cells' membrane potentials. Reversal potentials and membrane potentials shifted equally when cells were uniformly depolarized in 24 mM external K+. 3. Bath-applied dopamine elicited, after a delay of 4-9 min, an exaggerated form of the spontaneous behaviour that frequently gave way to a sudden large (up to thirtyfold) conductance change. At concentrations of 100-300 nM, a range of agonists with increasing affinity for the D3 receptor (apomorphine, quinpirole, 7-OH DPAT and PD 128907) triggered the response. The actions were neither mimicked by SKF-38393 nor antagonized by SCH-23390 (a selective D1 agonist and antagonist, respectively). Haloperidol reversibly blocked responses elicited by the D3/D2 agonist quinpirole. The action of effective agonists was maintained when transmitter release was abolished. Given the reported lack of D2 receptors in the islands of Calleja, these findings indicate a direct action of dopamine at the D3 receptor. 4. The dopaminergic effects were not affected by Gd3+ or substantial replacement of external Na+ with TEA, Tris or choline, eliminating stretch-activated channels but suggesting that if transmembrane channels were to be involved in this dopaminergic action they posseses a non-selective permeability to large cations. The reported presence of gap junctions in the islands of Calleja offers the explanation that these effects derive from enhanced activity of such channels or their hemi-constituents. 5. In testing the possible involvement of gap junctional coupling the following experimental observations were made: (i) alkalinization of slices mimicked the effect of D3 agonists; (ii) in cell groups, recording from pairs provided evidence of intercellular coupling, and mechanical separation of recorded neurones from neighbouring cells during the agonist-evoked response caused shutdown of the additional conductance; (iii) when applied to slices, the gap junctional blocker, 18 alpha-glycyrrhetinic acid, whilst not preventing the full-blown dopamine response, significantly reduced both the variance of recorded granule cell input conductance and the cells' apparent capacitance. 6. Taken together the results indicate a D3 action in granule cells, which is best explained by a dopaminergic promotion of intercellular coupling. The physiological relevance of such a mechanism is discussed.[1]


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