The PhD thesis addresses the analysis of GABA and glycine cotransmitting (GgC) neurons in the Pre-Bötzinger Complex (PBC), a region important for rhythm generation of the respiration. GgC neurons were identified in the PBC using different approaches including single-cell RT-PCR and immunohistochemistry. Furthermore, novel transgenic mouse-lines were generated to facilitate the analysis of these neurons in-vivo and in living-tissue preparations: The mouse-line Cofluor allows the identification of GABAergic, glycinergic as well as GgC neurons by the expression of different fluorescent proteins. This mouse-line was used to investigate the number of cells of each neuron type during development from E15.5 to 2.5 month revealing a decrease of GgC neurons paralleled by an increase of glycinergic and GABAergic neurons. The mouse line COTRIND expresses the tamoxifen-inducible Split-CreERT2 system exclusively in GgC neurons allowing for the permanent labelling of these neurons with the Red Fluorescent Protein at the time-point of tamoxifen application. In COTRIND mice, GgC neurons were irreversibly labelled at the age of P1/P2 and analysed at the age of three days as well as after 2.5 month to investigate their fate by using immunohistochemistry. It was found that GgC neurons differentiated to mostly glycinergic neurons and to a minor percentage to GABAergic neurons whereas some GgC neurons remained GgC neurons. The results obtained from this study disclosed the presence of GgC neurons in the PBC of mice and their development from the embryonic day 15.5 to the adult stage. Furthermore, the inducible Split-CreERT2 system was for the first time established in transgenic COTRIND mice and found to be functional in vivo without leakage. With the novel transgenic mouse lines the present work provides tools for the analysis of GgC neurons in the PBC and contributes to the characterisation of this neuron type including the role for rhythm generation of the respiration.

The PhD thesis addresses the analysis of GABA and glycine cotransmitting (GgC) neurons in the Pre-Bötzinger Complex (PBC), a region important for rhythm generation of the respiration. GgC neurons were identified in the PBC using different approaches including single-cell RT-PCR and immunohistochemistry. Furthermore, novel transgenic mouse-lines were generated to facilitate the analysis of these neurons in-vivo and in living-tissue preparations: The mouse-line Cofluor allows the identification of GABAergic, glycinergic as well as GgC neurons by the expression of different fluorescent proteins. This mouse-line was used to investigate the number of cells of each neuron type during development from E15.5 to 2.5 month revealing a decrease of GgC neurons paralleled by an increase of glycinergic and GABAergic neurons. The mouse line COTRIND expresses the tamoxifen-inducible Split-CreERT2 system exclusively in GgC neurons allowing for the permanent labelling of these neurons with the Red Fluorescent Protein at the time-point of tamoxifen application. In COTRIND mice, GgC neurons were irreversibly labelled at the age of P1/P2 and analysed at the age of three days as well as after 2.5 month to investigate their fate by using immunohistochemistry. It was found that GgC neurons differentiated to mostly glycinergic neurons and to a minor percentage to GABAergic neurons whereas some GgC neurons remained GgC neurons. The results obtained from this study disclosed the presence of GgC neurons in the PBC of mice and their development from the embryonic day 15.5 to the adult stage. Furthermore, the inducible Split-CreERT2 system was for the first time established in transgenic COTRIND mice and found to be functional in vivo without leakage. With the novel transgenic mouse lines the present work provides tools for the analysis of GgC neurons in the PBC and contributes to the characterisation of this neuron type including the role for rhythm generation of the respiration.

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