% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@PHDTHESIS{Kaschuba:14340,
author = {Kaschuba, Dagmar},
title = {{S}chrittmacherkanäle im olfaktorischen {E}pithel der
{M}aus},
volume = {4329},
issn = {0944-2952},
school = {Univ. Köln},
type = {Dr. (Univ.)},
address = {Jülich},
publisher = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
reportid = {PreJuSER-14340, Juel-4329},
series = {Berichte des Forschungszentrums Jülich},
pages = {X, 138 p.},
year = {2010},
note = {Record converted from VDB: 12.11.2012; Köln, Univ., Diss.,
2010},
abstract = {HCN channels hyperpolarization-activated and
$\underline{c}$yclic $\underline{n}$ucleotide-gated
channels) are membrane proteins participating in the
generation of spontaneous rhythmic electrical activity in
cellular networks. Therefore these channels are called
pacemaker channels. This thesis describes for the first
time, the expression pattern of four different HCN channel
isoforms (HCN1 - HCN4) in the olfactory epithelium of the
mouse on a subcellular level. The isoforms HCN1 and HCN2 are
expressed in olfactory receptor neurons (ORN), more
specifically in the dendrites and axons of the ORN. Strong
labeling was particularly seen in the axon bundles. The HCN4
isoform was found almost exclusively in the axons of the
ORN. Furthermore, the immunohistochemical stainings allowed
to distinguish between two morphologically different axon
bundles: small, tightly-packed axon bundles which express
HCN 1, 2 and 4 to similar degrees, and large axon bundles in
which predominantly HCN4 is expressed. Notably, HCN3 seems
not to be expressed in the olfactory epithelium of the
mouse. Specific shRNA molecules can be utilized to achieve a
post-transcriptional downregulation of genes. To this end,
$\underline{r}$ecombinant
$\underline{a}deno-$\underline{a}$ssociated
$\underline{v}$iruses (rAAV) were constructed allowing the
gene transfer of shRNA-coding sequences. In transgenic cell
lines which constitutively expressed specific HCN isoforms,
a significant downregulation of HCN1 and HCN2 gene
expression was achieved after infections with rAAV_shRNA
constructs. Especially for HCN2, de novo protein
biosynthesis was impaired almost completely. In a series of
experiments, the transduction capability of rAAV for ORN was
examined in vivo by virus-mediated eGFP expression. Both,
ORN and supporting cells were successfully transduced by
rAAV of serotype 2 and 5. In transduced cells, eGFP
expression was very high and allowed to unequivocally
identify the different cell types by their morphology. In
summary, this thesis demonstrates that rAAV-mediated gene
transfer is a versatile method that can be used both, to
introduce genes into living organisms as well as to
specifically knock down gene expression by rAAV_shRNA
thereby supporting the ultimate goal to study a proteins’
function in vivo. As likely candidates, individual HCN
isoforms might now be targeted as their subcellular
expression pattern has been unraveled in the olfactory
epithelium of the mouse.},
cin = {ISB-1},
cid = {I:(DE-Juel1)VDB922},
pnm = {BioSoft: Makromolekulare Systeme und biologische
Informationsverarbeitung},
pid = {G:(DE-Juel1)FUEK505},
typ = {PUB:(DE-HGF)11 / PUB:(DE-HGF)3},
url = {https://juser.fz-juelich.de/record/14340},
}