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Differential Expression of PKD2-Associated Genes in Autosomal Dominant Polycystic Kidney Disease.

Yook YJ, Woo YM, Yang MH, Ko JY, Kim BH, Lee EJ, Chang ES, Lee MJ, Lee S, Park JH - Genomics Inform (2012)

Bottom Line: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture.The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction.These data may be helpful in PKD2-related mechanisms of ADPKD pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Sookmyung Women's University, Seoul 140-742, Korea.

ABSTRACT
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture. The proteins encoded by the PKD1 and PKD2 genes, mutations in which account for nearly all cases of ADPKD, may help guard against cystogenesis. Previously developed mouse models of PKD1 and PKD2 demonstrated an embryonic lethal phenotype and massive cyst formation in the kidney, indicating that PKD1 and PKD2 probably play important roles during normal renal tubular development. However, their precise role in development and the cellular mechanisms of cyst formation induced by PKD1 and PKD2 mutations are not fully understood. To address this question, we presently created Pkd2 knockout and PKD2 transgenic mouse embryo fibroblasts. We used a mouse oligonucleotide microarray to identify messenger RNAs whose expression was altered by the overexpression of the PKD2 or knockout of the Pkd2. The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction. Herein, we confirmed differential expressions of several genes including aquaporin-1, according to different PKD2 expression levels in ADPKD mouse models, through microarray analysis. These data may be helpful in PKD2-related mechanisms of ADPKD pathogenesis.

No MeSH data available.


Related in: MedlinePlus

Expression verification of AQP1 at the protein level. (A) Western blot analysis of AQP1 in MEFs and kidney tissues derived from Pkd2 KO (or heterozygote) and PKD2 TG mice. PKD2 TG and TGWT mice: 18 mo; Pkd2 heterozygote and KOWT mice: 12 mo. Anti-β-actin was used as the internal control. (B) For quantification of western blot, Multigauge software (Fuji Film) was used. AQP1, aquaporin 1; MEF, mouse embryo fibroblasts; KO, knockout; TG, transgenic; KOWT, knockout wild-type (MEF); TGWT, transgenic wild-type (MEF).
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Figure 4: Expression verification of AQP1 at the protein level. (A) Western blot analysis of AQP1 in MEFs and kidney tissues derived from Pkd2 KO (or heterozygote) and PKD2 TG mice. PKD2 TG and TGWT mice: 18 mo; Pkd2 heterozygote and KOWT mice: 12 mo. Anti-β-actin was used as the internal control. (B) For quantification of western blot, Multigauge software (Fuji Film) was used. AQP1, aquaporin 1; MEF, mouse embryo fibroblasts; KO, knockout; TG, transgenic; KOWT, knockout wild-type (MEF); TGWT, transgenic wild-type (MEF).

Mentions: To verify the results obtained by cDNA microarray analysis, expression patterns for selected genes were confirmed at the RNA and protein levels. We selected seven genes among the 45 genes (Fig. 3A). As result of realtime RT-PCR, the expression patterns of PKCα, ITGα4, and AQP1 were identical to microarray analysis in Pkd2 KO MEF cells (Fig. 3B). In contrast, 6 genes, except for TGF-β2, were identically expressed in PKD2 TG MEF cells (Fig. 3C). Especially, the AQP1 gene may be significantly regulated by differential Pkd2 gene expression level. Furthermore, to confirm the AQP1 protein expression level, western blot analysis was performed using MEF cells as well as kidney tissues obtained from Pkd2 KO (or heterozygote) and PKD2 TG mice. As result, AQP1 protein level was consistent with mRNA levels in both MEFs and tissues (Fig. 4).


Differential Expression of PKD2-Associated Genes in Autosomal Dominant Polycystic Kidney Disease.

Yook YJ, Woo YM, Yang MH, Ko JY, Kim BH, Lee EJ, Chang ES, Lee MJ, Lee S, Park JH - Genomics Inform (2012)

Expression verification of AQP1 at the protein level. (A) Western blot analysis of AQP1 in MEFs and kidney tissues derived from Pkd2 KO (or heterozygote) and PKD2 TG mice. PKD2 TG and TGWT mice: 18 mo; Pkd2 heterozygote and KOWT mice: 12 mo. Anti-β-actin was used as the internal control. (B) For quantification of western blot, Multigauge software (Fuji Film) was used. AQP1, aquaporin 1; MEF, mouse embryo fibroblasts; KO, knockout; TG, transgenic; KOWT, knockout wild-type (MEF); TGWT, transgenic wild-type (MEF).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3475485&req=5

Figure 4: Expression verification of AQP1 at the protein level. (A) Western blot analysis of AQP1 in MEFs and kidney tissues derived from Pkd2 KO (or heterozygote) and PKD2 TG mice. PKD2 TG and TGWT mice: 18 mo; Pkd2 heterozygote and KOWT mice: 12 mo. Anti-β-actin was used as the internal control. (B) For quantification of western blot, Multigauge software (Fuji Film) was used. AQP1, aquaporin 1; MEF, mouse embryo fibroblasts; KO, knockout; TG, transgenic; KOWT, knockout wild-type (MEF); TGWT, transgenic wild-type (MEF).
Mentions: To verify the results obtained by cDNA microarray analysis, expression patterns for selected genes were confirmed at the RNA and protein levels. We selected seven genes among the 45 genes (Fig. 3A). As result of realtime RT-PCR, the expression patterns of PKCα, ITGα4, and AQP1 were identical to microarray analysis in Pkd2 KO MEF cells (Fig. 3B). In contrast, 6 genes, except for TGF-β2, were identically expressed in PKD2 TG MEF cells (Fig. 3C). Especially, the AQP1 gene may be significantly regulated by differential Pkd2 gene expression level. Furthermore, to confirm the AQP1 protein expression level, western blot analysis was performed using MEF cells as well as kidney tissues obtained from Pkd2 KO (or heterozygote) and PKD2 TG mice. As result, AQP1 protein level was consistent with mRNA levels in both MEFs and tissues (Fig. 4).

Bottom Line: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture.The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction.These data may be helpful in PKD2-related mechanisms of ADPKD pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Sookmyung Women's University, Seoul 140-742, Korea.

ABSTRACT
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture. The proteins encoded by the PKD1 and PKD2 genes, mutations in which account for nearly all cases of ADPKD, may help guard against cystogenesis. Previously developed mouse models of PKD1 and PKD2 demonstrated an embryonic lethal phenotype and massive cyst formation in the kidney, indicating that PKD1 and PKD2 probably play important roles during normal renal tubular development. However, their precise role in development and the cellular mechanisms of cyst formation induced by PKD1 and PKD2 mutations are not fully understood. To address this question, we presently created Pkd2 knockout and PKD2 transgenic mouse embryo fibroblasts. We used a mouse oligonucleotide microarray to identify messenger RNAs whose expression was altered by the overexpression of the PKD2 or knockout of the Pkd2. The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction. Herein, we confirmed differential expressions of several genes including aquaporin-1, according to different PKD2 expression levels in ADPKD mouse models, through microarray analysis. These data may be helpful in PKD2-related mechanisms of ADPKD pathogenesis.

No MeSH data available.


Related in: MedlinePlus