Abstract
Postnatal intestinal development is a very dynamic process characterized by substantial morphological changes that coincide with functional adaption to the nutritional change from a diet rich in fat (milk) to a diet rich in carbohydrates on from weaning. Time-resolved studies of intestinal development have so far been limited to investigation at the transcription level or to single or few proteins at a time. In the present study, we elucidate proteomic changes of primary intestinal epithelial cells from jejunum during early suckling (1-7 days of age), middle suckling (7-14 days), and weaning period (14-35 days) in mice, using a label-free proteomics approach. We show differential expression of 520 proteins during intestinal development and a pronounced change of the proteome during the middle suckling period and weaning. Proteins involved in several metabolic processes were found differentially expressed along the development. The temporal expression profiles of enzymes of the glycolysis were found to correlate with the increase in carbohydrate uptake at weaning, whereas the abundance changes of proteins involved in fatty acid metabolism as well as lactose metabolism indicated a nondiet driven preparation for the nutritional change at weaning. Further, we report the developmental abundance changes of proteins playing a vital role in the neonatal acquisition of passive immunity. In addition, different isoforms of several proteins were quantified, which may contribute to a better understanding of the roles of the specific isoforms in the small intestine. In summary, we provide a first, time-resolved proteome profile of intestinal epithelial cells along postnatal intestinal development.
Highlights
The postnatal development of the gastrointestinal tract is a very dynamic process, including an intensive growth together with a complex process of differentiation
Establishment of Label-free Quantitative Proteomics of Intestinal Development—The goal of this study was to elucidate proteomic changes of intestinal epithelial cells (IECs) along postnatal maturation of the murine intestine. We approached this with a label-free, quantitative proteomics method using four time-points along postnatal maturation to analyze the changes occurring at the three periods of early suckling (1–7 D), middle suckling (7–14 D), and weaning period (14 –35 D) (Fig. 1)
Based on quantitative proteomics of primary IECs, we have generated a unique and extensive data set of protein abundance changes that define in vivo intestinal development
Summary
The postnatal development of the gastrointestinal tract is a very dynamic process, including an intensive growth together with a complex process of differentiation. Long and thin villi shape the small intestinal epithelium, and the enterocytes (the main absorptive cells of the intestine) are characterized by their apical microvilli [6]. Before birth in humans and during the first week of age in mice, crypts develop from the intervillous region, resulting in the formation of a distinct proliferating compartment [6]. Final intestinal development in the postweaning phase is characterized by growth of crypts and villi, which results in an expansion of the villi from a finger-like to a leaflike shape [7]. Between the three mentioned phases of intestinal development, two abrupt changes in diet occur: at birth, when the intestine switches from processing dilute amniotic fluid to digesting milk; and at weaning, when the adaption to solid food begins. The mammalian intestinal tract is rapidly colonized by bacteria from the mother and the surrounding environment, eventually resulting in a dense and diverse in-
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