Voltage-gated K+ channel (Kv) subunit expression of the guinea pig spiral ganglion cells studied in a newly developed cochlear free-floating preparation.

The spiral ganglion accommodates the cell bodies of the acoustic nerve fibres connecting the hair cells to the central nervous system. As the ionic channels containing various voltage-gated K+ channel (Kv) subunits play pivotal roles in determining the functional properties and firing behaviour of the spiral ganglion cells (SGCs), every piece of information concerning the Kv expression of the SGCs is valuable. In the present work a comprehensive immunohistochemical analysis was performed to describe the expression of 9 Kv subunits in the guinea pig cochlea on traditional wax-embedded sections as well as employing a newly developed preparation that allowed confocal analysis, reconstruction of the three-dimensional appearance and precise morphological characterisation of the SGCs. Besides determining their Kv expression patterns, differences between type I and type II SGCs were sought. SGCs showed positivity for 8 out of the 9 Kv subunit-specific antibodies with varying intensity and proportion of the immunopositive cells; whereas no obvious Kv3.2 positivity could be noted. Type I and type II cells demonstrated similar expression patterns for all subunits tested, with the exception of Kv1.2, whose presence was confirmed in only 50% of the type II cells. Although the present findings suggest that type I and type II cells do not differ fundamentally in the Kv subunits they possess; they also imply that SGCs may not form a homogeneous cell population, and might provide explanation of the previously noted heterogeneity of the membrane properties of the SGCs.

Voltage-gated potassium channel (Kv) subunits expressed in the rat cochlear nucleus.

Because the neuronal membrane properties and firing characteristics are crucially affected by the depolarization-activated K(+) channel (Kv) subunits, data about the Kv distribution may provide useful information regarding the functionality of the neurons situated in the cochlear nucleus (CN). Using immunohistochemistry in free-floating slices, the distribution of seven Kv subunits was described in the rat CN. Positive labeling was observed for Kv1.1, 1.2, 1.6, 3.1, 3.4, 4.2, and 4.3 subunits. Giant and octopus neurons showed particularly strong immunopositivity for Kv3.1; octopus neurons showed intense Kv1.1- and 1.2-specific reactions also. In the latter case, an age-dependent change of the expression pattern was also documented; although both young and older animals produced definite labeling for Kv1.2, the intensity of the reaction increased in older animals and was accompanied with the translocation of the Kv1.2 subunits to the cell surface membrane. The granule cell layer exhibited strong Kv4.2-specific immunopositivity, and markedly Kv4.2-positive glomerular synapses were also seen. It was found that neither giant nor pyramidal cells were uniform in terms of their Kv expression patterns. Our data provide new information about the Kv expression of the CN and also suggest potential functional heterogeneity of the giant and pyramidal cells.

Mitochondrial expression of the two-pore domain TASK-3 channels in malignantly transformed and non-malignant human cells.

The presence of TASK-3 channels has been described in a number of healthy and malignantly transformed cells, showing mainly intracellular distribution with relatively insignificant labelling of the cell surface membrane. In this work, immunochemical and molecular biology methods were utilised to establish the intracellular organelle whose TASK-3 expression accounts for this strong intracellular labelling using cultured melanoma and HaCaT cells. Before the immunocytochemical experiments, the presence of TASK-3 mRNA was also confirmed in melanoma cells. Comparison of the results of the TASK-3- and mitochondrion-specific labelling indicated that the TASK-3 channel subunits were strongly expressed by mitochondria in both investigated cell types. Moreover, prominent TASK-3 expression of keratinocytes could also be demonstrated in histological sections excised from the human skin. These results indicate that TASK-3 channels are present in the mitochondria in both malignantly transformed and healthy cells, suggesting that they might have roles in ensuring mitochondrial functions.

Rhodamine backfilling and confocal microscopy as a tool for the unambiguous identification of neuronal cell types: a study of the neurones of the rat cochlear nucleus.

Adequate interpretation of the functional data characterising the projection neurones of the cochlear nucleus (CN) is impossible without the unequivocal classification of these cell types at the end of the experiments. In this study, morphological criteria applicable for unambiguous identification of CN neurones have been sought. The neurones were labelled with rhodamine from incisions severing the projection pathways of the individual cell types, allowing their selective labelling and morphological characterisation. Confocal microscopy was employed for the investigation of the rhodamine-filled cells whose morphology was assessed after reconstructing the three-dimensional images of the cell bodies and proximal processes. The diameters of the somata and the number of processes originating from the cell bodies were also determined. In most of the cases, unambiguous identification of the bushy, octopus and Purkinje-like cells was relatively straightforward. On the other hand, precise classification of the pyramidal cells was often difficult, especially because giant cells could easily possess morphological features resembling pyramidal neurones. Occasionally, giant cells also mimicked the appearance of octopus neurones, which may be another important source of identification error, especially as these two cell types are often situated close to each other in the CN. It is concluded that morphological criteria defined in the present work may be effectively applied for the unambiguous identification of the projection neurones of the CN, even following functional measurements, when the correct cell classification is essential for the interpretation of the experimental data. Moreover, the present study also confirmed that Purkinje-like cells project to the cerebellum.