Because the measurement of α7 expression and function can be comp

Because the measurement of α7 expression and function can be compromised by low receptor

expression levels or the absence of conditions that best reveal its modulatory role (Gahring and Rogers 2005; Albuquerque et al. 2009), the participation by this receptor as an important contributor to the development and normal auditory sensory function remains to be fully explored. In this study, we examine α7 expression during development of the auditory sensory system. This was done using mice that were modified though methods Inhibitors,research,lifescience,medical of homologous recombination (Rogers and Gahring 2012; Rogers et al. 2012) to introduce, at the α7 gene 3′ end, a hemagglutinin (HA) protein tag to the α7 receptor protein and a bicistronic IRES-driven tau + enhanced-GFP Inhibitors,research,lifescience,medical fusion protein reporter (α7GFP). An advantage of the tauGFP reporter construct

is that the tau component directs GFP into the axon of cells expressing α7GFP. Also generated was a mouse in which Cre-recombinase replaces the tauGFP. The expression of α7GFP in these mice reveals extensive spatial and temporal remodeling of receptor expression during embryonic and postnatal development of the cochlear sensory structures. Furthermore, α7GFP expression continues in both neuronal and non-neuronal cells of the adult cochlear structure and the central ascending auditory pathway. This suggests that α7 has the potential to impact functionally on auditory processes through multiple pathways and mechanisms Inhibitors,research,lifescience,medical that could impact upon the adult function in ways not traditionally attributed to this receptor. Materials and Methods Animals All animals were used and housed in accordance with protocols approved in advance by the Institutional Animal Care and Use Committee at the University of Utah (Protocol Number (selleck inhibitor 09-07003). This includes adherence to the Inhibitors,research,lifescience,medical Guide for the Care and use of Laboratory Animals of the National Institutes of Health. Generation of alpha7-HA-IRES-tauGFP and alpha7-HA-IRES-Cre Inhibitors,research,lifescience,medical mice The Gemcitabine synthesis construction of the α7 protein and gene (Chrna7) reporter mouse lines; Chrna7-HA-IRES-tauGFP (α7GFP)

and Chrna7-HA-IRES-Cre (α7Cre) have been described in detail (Rogers and Gahring 2012; Rogers Dacomitinib et al. 2012). Briefly, as diagramed in Fig. 1A, the methods of homologous recombination were used to introduce an epitope hemagglutinin (HA) and stop codon extension to the α7 C-terminus and a bicistronic IRES-tauGFP reporter cassette (Rogers and Gahring 2012; Rogers et al. 2012). This generated the α7GFP mouse (Fig. 1A), which expresses the tauGFP protein as a marker of Chrna7 transcription. The Speed Congenic Program of the Jackson Laboratory was used to achieve 98% C57BL/6 background congenicity (Rogers et al. 2012). For conditional cell ablation of the cells expressing Cre as in the α7Cre mouse, we crossed this mouse with the LoxP conditional diphtheria toxin (DTA) mouse lines as described previously (Rogers et al. 2012). Figure 1 Mouse models used to examine nicotinic receptor α7 expression.

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