Effect of zinc deficiency on the mRNA expression pattern in liver and jejunum of adult rats: monitoring gene expression using cDNA microarrays combined with real-time RT-PCR.
In the study presented here, the effect of zinc deficiency on mRNA expression levels in liver and jejunum of adult rats was analyzed. Feed intake was restricted to 8 g/day. The semi-synthetic diet was fortified with pure phytate and contained either 2 microg Zn/g (Zn deficiency, n = 6) or 58 microg Zn/g (control, n = 7). After 29 days of Zn depletion feeding, entire jejunum and liver were retrieved and total RNA was extracted. Tissue specific expression pattern were screened and quantified by microarray analysis and verified individually via real-time RT-PCR. A relative quantification was performed with the newly developed Relative Expression Software Tool Copyright on numerous candidate genes which showed a differential expression. This study provides the first comparative view of gene expression regulation and fully quantitative expression analysis of 35 candidate genes in a non-growing Zn deficient adult rat model. The expression results indicate the existence of individual expression pattern in liver and jejunum and their tissue specific regulation under Zn deficiency. In addition, in jejunum a number of B-cell related genes could be demonstrated to be suppressed at Zn deficiency. In liver, metallothionein subtype 1 and 2 (MT-1 and MT-2) genes could be shown to be dramatically repressed and therefore represent putative markers for Zn deficiency. Expression results imply that some genes are expressed constitutively, whereas others are highly regulated in tissues responsible for Zn homeostasis.
Pfaffl
Michael W
MW
Institute of Physiology, Department of Animal Sciences, Centre of Life and Food Sciences, Technical University of Munich, 85354 Freising, Germany. pfaffl@wzw.tum.de
Gerstmayer
B
B
Bosio
A
A
Windisch
Wilhelm
W
D000818:Animals; D001402:B-Lymphocytes; D005260:Female; D015870:Gene Expression; D007583:Jejunum / Q000737:chemistry*; D008099:Liver / Q000737:chemistry*; D008668:Metallothionein / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015032:Zinc / Q000172:deficiency*
D012333:RNA, Messenger; D012313:RNA; D008668:Metallothionein; D015032:Zinc