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J Physiol Volume 560, Number 1, 249-266, October 1, 2004 DOI: 10.1113/jphysiol.2004.070540
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Physiological, neurochemical and morphological properties of a subgroup of GABAergic spinal lamina II neurones identified by expression of green fluorescent protein in mice

Bernhard Heinke1, Ruth Ruscheweyh1, Liesbeth Forsthuber1, Gabriele Wunderbaldinger1 and Jürgen Sandkühler1

1 Centre for Brain Research, Department of Neurophysiology, Medical University Vienna, Vienna, Austria

The processing of sensory, including nociceptive, information in spinal dorsal horn is critically modulated by spinal GABAergic neurones. For example, blockade of spinal GABAA receptors leads to pain evoked by normally innocuous tactile stimulation (tactile allodynia) in rats. GABAergic dorsal horn neurones have been classified neurochemically and morphologically, but little is known about their physiological properties. We used a transgenic mouse strain coexpressing enhanced green fluorescent protein (EGFP) and the GABA-synthesizing enzyme GAD67 to investigate the properties of a subgroup of GABAergic neurones. Immunohistochemistry showed that EGFP-expressing neurones accounted for about one-third of the GABAergic neurones in lamina II of the spinal dorsal horn. They constituted a neurochemically rather heterogeneous group where 27% of the neurones coexpressed glycine, 23% coexpressed parvalbumin and 14% coexpressed neuronal nitric oxide synthase (nNOS). We found almost no expression of protein kinase C{gamma} (PKC{gamma}) in EGFP-labelled neurones but a high costaining with PKCßII (78%). The whole-cell patch-clamp technique was used to intracellularly label and physiologically characterize EGFP- and non-EGFP-expressing lamina II neurones in spinal cord slices. Sixty-two per cent of the EGFP-labelled neurones were islet cells while the morphology of non-EGFP-labelled neurones was more variable. When stimulated by rectangular current injections, EGFP-expressing neurones typically exhibited an initial bursting firing pattern while non-EGFP-expressing neurones were either of the gap or the delayed firing type. EGFP-expressing neurones received a greater proportion of monosynaptic input from the dorsal root, especially from primary afferent C-fibres. In conclusion, EGFP expression defined a substantial but, with respect to the measured parameters, rather inhomogeneous subgroup of GABAergic neurones in spinal lamina II. These results provide a base to elucidate the functional roles of this subgroup of GABAergic lamina II neurones, e.g. for nociception.

(Received 23 June 2004; accepted after revision 28 July 2004; first published online 29 July 2004)
Corresponding author J. Sandkühler: Centre for Brain Research, Department of Neurophysiology, Medical University Vienna, Spitalgasse 4, A-1090 Vienna, Austria. Email: juergen.sandkuehler{at}meduniwiet.ac.at


B. Heinke and R. Ruscheweyh contributed equally to this work




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