This study assessed the effects of enhanced dietary plane of nutrition (early nutritional program (ENH)) on the gene expression pattern of ruminal epithelial tissue of young Holstein calves. Male Holstein calves were fed (3 to 42 days of age) with reconstituted control milk replacer (MR) (20 % crude protein, 20 % fat; 1.25 lb solids/calf) plus conventional starter (CON; 19.6 % crude protein, dry matter basis) or a high-protein MR (ENH; 28.5 % crude protein, 15 % fat; at around 2 % of body weight) plus high-crude protein starter (25.5 % crude protein, dry matter basis). The calves were weaned on day 43. Groups of calves in CON and ENH treatment were harvested after 5 and 10 weeks of feeding. The ruminal epithelium from five calves in each group was used for transcript profiling using a bovine oligonucleotide microarray. The postweaning mass of the reticulo-rumen was greater (P < 0.01) in calves consuming ENH. Transcriptome analysis revealed that 208 genes were altered due to treatment and 587 due to time alone. Bioinformatics analysis revealed that "galactose metabolism," "citrate cycle," "pyruvate metabolism," and "basal transcription factors" were the most impacted and induced pathways due to feeding ENH; whereas, "valine, leucine, and isoleucine biosynthesis" and "glyoxylate and dicarboxylate metabolism" were among the most inhibited. The integrated interpretation of the results suggested an overall increase in metabolism after weaning, particularly biosynthesis of glycan and nucleotide metabolism. Furthermore, the preweaning alterations in the transcriptome were mostly associated with cell growth, death, tissue development, and cellular morphology. The postweaning response revealed overexpression of genes associated with cell adhesion molecules, p53 signaling, and fatty acid metabolism. Our results indicated that feeding ENH to young Holstein calves elicited a strong transcriptomic response in the ruminal epithelial tissue.