Research article
Global effects of vitamin A deficiency on gene expression in rat liver: evidence for hypoandrogenism

https://doi.org/10.1016/j.jnutbio.2005.08.006Get rights and content

Abstract

Vitamin A (retinol) metabolites are ligands for transcription factors that regulate many genes. The liver is the main storage depot for retinol and plays a role in vitamin A homeostasis. To better understand the effects of vitamin A deficiency on liver gene expression, we produced retinol deficiency in male rats by feeding a diet low in retinol for 53 days after weaning and examined the effects on gene expression in liver using Affymetrix oligonucleotide microarrays. We detected expression of 41% of the 8799 probe sets represented on the RGU-34A GeneChips. Vitamin A deficiency resulted in major changes in liver gene expression: 805 genes (22% of all genes detected) differed at P≤.05 (false discovery rate <0.143). Genes involved in fatty acid metabolism, peroxisomal function, glycolysis, glutamate metabolism and the urea cycle were altered. The expression of many sexually dimorphic genes was altered toward a feminized or senescent pattern of gene expression in the liver. Retinol deficiency also produces a shift toward increased protein and fat catabolism and decreased fatty acid synthesis.

Introduction

Vitamin A (retinol) plays an important role in many physiological activities through its metabolites, particularly 9-cis and all-trans retinoic acids. These retinoids are ligands for important transcription factors, including the classical all-trans retinoic acid receptors (RARs) and the 9-cis retinoic acid receptors (retinoid X receptors, RXRs). RXRs form heterodimers with many other transcription factors, including the vitamin D receptor, peroxisome proliferator-activated receptors (PPARs), thyroid hormone receptors, liver X receptor, farnesoid X receptor, pregnane X receptor and the constitutive androstane receptor [1]. These transcription factors, in turn, regulate the expression of a large number of genes. Thus, vitamin A deficiency is expected to have a major impact on gene expression.

Vitamin A deficiency has wide-ranging effects, among which are visual problems, increased inflammation, decreased growth, and, in males, decreased secretion of testosterone and blocked spermatogenesis [2]. In the liver, vitamin A deficiency has a large effect on the total microsomal content of cytochrome P450s [3] and is associated with decreased expression of insulin-like growth factor 1 (IGF-1), leading to decreased circulating levels of IGF-1 [4]. Decreased IGF-1 levels are also seen in starvation and calorie or diet restriction [5], [6]. The effects of diet and vitamin A deficiency on IGF-1 may be related because vitamin A deficiency changes enzyme levels and histology of the small intestine accompanied by increased inflammation that can lead to decreased absorption of nutrients and decreased growth [7]. The liver is the major storage organ for retinol and is involved in most aspects of vitamin A homeostasis. Most retinol is stored in the hepatic stellate cells as retinyl-palmitate esters [8]. The liver expresses the retinoic acid-regulated transcription factors, with the exception of the vitamin D receptor, at high levels [9].

To understand the global effects of vitamin A deficiency on gene expression in liver, we have examined the mRNA expression profile using Affymetrix oligonucleotide microarrays.

Section snippets

Induction of vitamin A deficiency in rats

Male Sprague–Dawley rats were made deficient in vitamin A by feeding a custom vitamin A-deficient diet based on the AIN93 composition (ICN, Costa Mesa, CA). They were compared with rats fed the same diet but containing 4000 IU vitamin A palmitate/kg of chow (vitamin A-sufficient). Both groups of rats were fed ad libitum. The diets were started at 21 days of age. The animals were sacrificed on the morning of Day 53 of the diet. These animals were described as part of an earlier study on the

Global effects of retinol deficiency on rat liver

Three-week-old weanling male Sprague–Dawley rats were fed a custom vitamin A-deficient or vitamin A-sufficient diet for 53 days. This resulted in progressive vitamin A deficiency as the vitamin A stores present at birth and from maternal milk were depleted. As previously reported [10], growth of the vitamin A-deficient animals slowed after 31 days on the diet, suggesting the animals were vitamin-deficient for about 3 weeks before sacrifice. Total retinol in serum was at or near zero (0.5±1.6

Discussion

One of the strengths of gene expression profiling is that patterns of change may be recognized that would be difficult to understand by analyzing individual mRNAs in isolation. This experiment revealed that vitamin A deficiency resulted in major changes in liver gene expression: 22% of all genes whose expression was detected demonstrated altered expression. We have provided a supplemental table containing all of the probe sets with a P≤.05 and have deposited all of the data in the GEO database

Acknowledgments

This work was supported by grants to DWC (AA06434), JAP (AA07462 training grant), the Indiana Alcohol Research Center (AA07611), the Indiana 21st Century Research and Technology Fund (HJE) and the Indiana Genomics Initiative (INGEN is supported in part by the Lilly Endowment).

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