Project description:This is a study of the change in gene expression in mouse kidney after feeding control (1.0% P) or low phosphate diet (0.03% P) for 3 or 5 days to normal or Hyp (X-linked hypophosphatemic) mice at 5 weeks of age. The mice were C57BL/6J. Normal wild-type mice, hemizygous Hyp (Hyp/Y) male mice, or heterozygous (Hyp/+) female mice were used. The gene is dominant, so that the two genders are equally affected with low renal retention of phosphate and severity of their bone disease. RNA from 3 mice were pooled for each microarray. The arrays were processed as described in the Affymetrix GeneChip Expression Analysis Technical Manual (copyright 2001, Affymetrix, Inc., Santa Clara, CA), rev. 1, Part number 701021. The data for each array were scaled to an average signal value of 500 for all genes on the array. Keywords = mouse Keywords = kidney Keywords = low phosphate diet Keywords = Hyp Keywords = Phex Keywords = X-linked hypophosphatemia Keywords: repeat sample
Project description:The renal adaptation to changing dietary phosphate levels is not well understood. The dominant Hyp mutation of the Phex gene in mice causes X-linked hypophosphatemia with low renal retention of phosphate and blocks physiologic adaptation to low phosphate diets. At P < 0.01, there were 1,711 transcripts significantly affected by genotype, 1,428 by diet and 5,601 by sex. Many renal transporters other than phosphate, as well as many novel transcripts of unknown function, were affected by the Hyp mutation. Some genes for fat metabolism and inflammation were up-regulated in Hyp kidneys. Of the genes affected by genotype and diet, only 378 were affected by both. In summary, the Hyp mutation induced changes in mRNA levels for numerous transcripts exceeding that required to alter phosphate retention. The data suggest broader physiological roles for the Phex gene unrelated to phosphate conservation. Keyword = Phex; Keyword = Hyp; Keyword = mouse; Keyword = kidney; Keyword = sex; Keyword = mRNA; Keyword = microarray; Keyword = low phosphorus diet Experiment Overall Design: A 2x2x2 factorial design, balanced for genotype (normal vs. Hyp), diet (control vs. low phosphate), and sex (male vs. female) was employed. Control or low phosphate diets were fed for three days to 10-week-old mice. A total of 24 samples of renal RNA were collected from 72 mice (3/array), processed to biotin-labeled cRNA, and hybridized to Affymetrix mouse MOE 430A and 430B for measurement of expression of over 45,000 transcripts.
Project description:This is a study of the change in gene expression in mouse kidney after feeding control (1.0% P) or low phosphate diet (0.03% P) for 3 or 5 days to normal or Hyp (X-linked hypophosphatemic) mice at 5 weeks of age. The mice were C57BL/6J. Normal wild-type mice, hemizygous Hyp (Hyp/Y) male mice, or heterozygous (Hyp/+) female mice were used. The gene is dominant, so that the two genders are equally affected with low renal retention of phosphate and severity of their bone disease. RNA from 3 mice were pooled for each microarray. The arrays were processed as described in the Affymetrix GeneChip Expression Analysis Technical Manual (copyright 2001, Affymetrix, Inc., Santa Clara, CA), rev. 1, Part number 701021. The data for each array were scaled to an average signal value of 500 for all genes on the array.
Project description:The renal adaptation to changing dietary phosphate levels is not well understood. The dominant Hyp mutation of the Phex gene in mice causes X-linked hypophosphatemia with low renal retention of phosphate and blocks physiologic adaptation to low phosphate diets. At P < 0.01, there were 1,711 transcripts significantly affected by genotype, 1,428 by diet and 5,601 by sex. Many renal transporters other than phosphate, as well as many novel transcripts of unknown function, were affected by the Hyp mutation. Some genes for fat metabolism and inflammation were up-regulated in Hyp kidneys. Of the genes affected by genotype and diet, only 378 were affected by both. In summary, the Hyp mutation induced changes in mRNA levels for numerous transcripts exceeding that required to alter phosphate retention. The data suggest broader physiological roles for the Phex gene unrelated to phosphate conservation. Keyword = Phex Keyword = Hyp Keyword = mouse Keyword = kidney Keyword = sex Keyword = mRNA Keyword = microarray Keyword = low phosphorus diet Keywords: disease state analysis
Project description:Phosphate homeostasis is a known critical function performed by the mammalian kidneys. However, changes to the transcriptome of the mammalian kidney in response to phosphate loading is unclear. The goal of this study was to examine kidney gene expression in response to high phosphate diet, compared to normal phosphate diet. RNA-seq was performed on the kidneys of C57BL/6J mice who were fed a high phosphate diet and in mice who received a normal phosphate diet for three days.
Project description:The pathophysiology of the osteomalacia in X-linked hypophosphatemia is uncertain. In this project, genomic DNA microarrays were used to identify novel genes with abnormal mRNA expression levels in mice with the dominant Hyp mutation of the Phex gene. Femoral shafts from five-week-old C57BL/6J mice, male and female, normal and Hyp (hemizygous male and heterozygous female), were flushed with saline to remove the marrow. RNA was extracted from each bone, pooled between two mice for each array, processed to cRNA, and hybridized to Affymetrix Mouse 430 2.0 GeneChip microarrays with probe sets for 45,101 genes. Twenty microarrays (40 mice) were done with 5 arrays for each treatment group (normal male, Hyp male, normal female and Hyp female). For each gene, factorial analysis of variance was performed for the main effects of genotype (normal vs. Hyp), sex (male vs. female), and genotype-by-sex interaction. The mRNA levels for 54 % of the genes on each array were scored as present. At P < 0.01, 2,635 genes were significant for genotype, 1,488 for sex, and 509 for genotype-by-sex interaction. There were two probes sets for the Phex gene. Probe 1450445, at the 3â end of the coding sequence, was low in normal samples (246 ± 37 (10), mean ± SEM (n)) and absent in Hyp samples. Probe 1421979, at the far 3â end of the untranslated region of the cDNA, 3,000 base pairs from the coding sequence, was high in normal mice (3,915 ± 315 (10); 8x brighter than the average gene), undetectable in Hyp males, and 725 ± 93 (5) in Hyp females. Both probe sets were scored as absent in kidney tissue. In Hyp bone, male and female, there was significant down-regulation of markers of osteoblasts and bone matrix synthesis with significant up-regulation of markers of blood vessel formation and cytoskeleton. No prominent skeletal gene was up-regulated in Hyp to attempt to compensate for the low skeletal mineralization. The genes with significant genotype-by-sex interaction did not show a marked fold difference between male and female Hyp mice. In conclusion, male and female Hyp mice showed similar depression of mRNA levels of genes related to bone synthesis in the femoral shaft. There was a high signal level from probes for a sequence in the 3â untranslated region of the Phex gene of normal, but not Hyp, mice, suggesting the need for further study of the molecular organization of this gene. Experiment Overall Design: Equal amounts of RNA from two mice, matched for genotype and sex were pooled to create each sample for microarray analysis. Four treatment groups were done: (1) Normal male mice, (2) Hyp male mice, (3) normal female mice, and (4) Hyp female mice. Five replicates were done with each replicate containing one sample from each of the four treatment groups for a total of 20 independent samples (40 mice total). Each replicate was matched for littermates and parallel processing.
Project description:The pathophysiology of the osteomalacia in X-linked hypophosphatemia is uncertain. In this project, genomic DNA microarrays were used to identify novel genes with abnormal mRNA expression levels in mice with the dominant Hyp mutation of the Phex gene. Femoral shafts from five-week-old C57BL/6J mice, male and female, normal and Hyp (hemizygous male and heterozygous female), were flushed with saline to remove the marrow. RNA was extracted from each bone, pooled between two mice for each array, processed to cRNA, and hybridized to Affymetrix Mouse 430 2.0 GeneChip microarrays with probe sets for 45,101 genes. Twenty microarrays (40 mice) were done with 5 arrays for each treatment group (normal male, Hyp male, normal female and Hyp female). For each gene, factorial analysis of variance was performed for the main effects of genotype (normal vs. Hyp), sex (male vs. female), and genotype-by-sex interaction. The mRNA levels for 54 % of the genes on each array were scored as present. At P < 0.01, 2,635 genes were significant for genotype, 1,488 for sex, and 509 for genotype-by-sex interaction. There were two probes sets for the Phex gene. Probe 1450445, at the 3’ end of the coding sequence, was low in normal samples (246 ± 37 (10), mean ± SEM (n)) and absent in Hyp samples. Probe 1421979, at the far 3’ end of the untranslated region of the cDNA, 3,000 base pairs from the coding sequence, was high in normal mice (3,915 ± 315 (10); 8x brighter than the average gene), undetectable in Hyp males, and 725 ± 93 (5) in Hyp females. Both probe sets were scored as absent in kidney tissue. In Hyp bone, male and female, there was significant down-regulation of markers of osteoblasts and bone matrix synthesis with significant up-regulation of markers of blood vessel formation and cytoskeleton. No prominent skeletal gene was up-regulated in Hyp to attempt to compensate for the low skeletal mineralization. The genes with significant genotype-by-sex interaction did not show a marked fold difference between male and female Hyp mice. In conclusion, male and female Hyp mice showed similar depression of mRNA levels of genes related to bone synthesis in the femoral shaft. There was a high signal level from probes for a sequence in the 3’ untranslated region of the Phex gene of normal, but not Hyp, mice, suggesting the need for further study of the molecular organization of this gene. Keywords: 2x2 factorial design with complete blocks
Project description:At present, the global obesity problem is becoming more and more serious, puts a huge burden on individuals and society. It is imperative to lose weight. Lifestyle intervention is the most economical and least side-effect intervention method, and diet control is an important link. We designed a set of experiments to study the effect of low-fat diet on kidney complications in obese mice, and found the target protein of low-fat diet to protect the obesity-related glomerulopathy (ORG) and Kidney Renal Clear Cell Carcinoma (KIRC).