Functional genomic characterisation of the Drosophila melanogaster alkali-metal/proton exchanger (NHE) family
Başlık:
Functional genomic characterisation of the Drosophila melanogaster alkali-metal/proton exchanger (NHE) family
Yazar:
Giannakou, Maria Eleni, author.
ISBN:
9780438060159
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (249 pages)
Genel Not:
Source: Dissertation Abstracts International, Volume: 76-08C.
Advisors: Julian Dow.
Özet:
Na+/H+ exchangers are a vital component of the cell ionic regulation machinery, involved in cell volume regulation, pH homeostasis and ion transport. There are at least seven human NHE genes, five of which are plasma membrane (NHEl-5) and two are organellar (NHE6 and NHE7). Similar genes have been identified in various other organisms, such as other mammals, C.elegans, Xenopus, plants, yeast and bacteria. In insects, NHEs may have another significant role. According to the "Wieczorek model" for ion transport, some insect epithelia are energised by a H+ V-ATPase, which in turn drives an amiloride-sensitive alkali-metal/proton antiport. The Drosophila Malpighian tubule was chosen as a model epithelium to characterise this family of proteins in insects. Firstly, this project focuses on the amiloride sensitivity of the Malpighian tubule. A panel of amiloride analogues, were assayed for their effect on the Malpighian tubule using fluid secretion assays. The pharmacological sensitivity of the Drosophila Malpighian tubule to these drugs is consistent with an effect on an exchanger rather than a Na+ channel. Furthermore, no Na+ channel genes appear to be expressed in the Malpighian tubule. Secondly, this study describes the identification of three Drosophila members of the Na+/H+ exchanger protein (NHE) family by homology screening, DmNHEl, DmNHE2 and DmNHE3. The expression of these genes was mapped by RT-PCR and immunocytochemistry. All three genes are widely expressed, including expression in the Malpighian tubule, and are expressed at all developmental stages. Immunocytochemistry shows that DmNHE2 is primarily localised to the apical membrane, whereas the other two DmNHEs are likely to be organellar. Their putative translations mark them as distantly related members of the NHE family, and different Drosophila NHEs are more similar to different members of the NHE family. Lastly this project describes various approaches undertaken to study DmNHE function using reverse genetics and P element mutagenesis. A heritable RNAi approach was used to investigate the effect of silencing of these genes. Targetted overexpression of the DmNHEs using the GAL4AJAS system was used to analyse the function of each exchanger.
Notlar:
School code: 0547
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Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(684792.1) | 684792-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
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