Abstract

ᅟStarch requires six enzymes for digestion to free glucose: two amylases (salivary and pancreatic) and four mucosal maltase activities; sucrase-isomaltase and maltase-glucoamylase. All are deficient in suckling rodents.ObjectiveThe objective of this study is to test 13C-starch digestion before weaning by measuring enrichment of blood 13C-glucose in maltase-glucoamylase-null and wild-type mice.MethodsMaltase-glucoamylase gene was ablated at the N-terminal. Dams were fed low 13C-diet and litters kept on low 13C-diet. Pups were weaned at 21 days. Digestion was tested at 13 and 25 days by intragastric feeding of amylase predigested 13C-α-limit dextrins. Blood 13C-glucose enrichment was measured by gas chromatography combustion isotope ratio mass spectrometry (GCRMS) using penta-acetate derivatives.ResultsFour hours after feeding, blood 13C-glucose was enriched by 26 × 103 in null and 18 × 103 in wild-type mice at 13 days and 0.3 × 103 and 0.2 × 103 at 25 days (vs. fasting p = 0.045 and p = 0.045). By jejunal enzyme assay, immunohistochemistry, or Western blots, there was no maltase activity or brush border staining with maltase-glucoamylase antibodies at 13 days, but these were fully developed in the wild-type mice by 25 days. In 13-day null mice, luminal contents were stained by maltase-glucoamylase antibodies. Lactating the mammary gland revealed maltase-glucoamylase antibody staining of alveolar cells. Reverse transcription/polymerase chain reaction (RT/PCR) of lactating glands revealed a secreted form of maltase-glucoamylase.Conclusions(1) 13C-α-limit dextrins were rapidly digested to 13C-glucose in 13-day mice independent of maltase-glucoamylase genotype or mucosal maltase activity. (2) This experiment demonstrates that a soluble maltase activity is secreted in mouse mother’s milk which enables suckling pup starch digestion well before brush border enzyme development. (3) This experiment with 13C-α-limit dextrins needs to be repeated in human breast fed infants.

Highlights

  • Maltase-glucoamylase gene was ablated at the N-terminal

  • Four hours after 13C-limit dextrins (LDx) feeding, blood 13C-glucose was enriched by 26 × 103 in null and 18 × 103 in WT at 13 days and 0.3 × 103 and 0.2 × 103 at 25 days

  • The tissue was homogenized in phosphate-buffered saline (PBS) buffer with ethylene diamine tetra-acetic acid (EDTA)

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Summary

Methods

Maltase-glucoamylase gene was ablated at the N-terminal. Dams were fed low 13C-diet and litters kept on low 13C-diet. Blood 13C-glucose enrichment was measured by gas chromatography combustion isotope ratio mass spectrometry (GCRMS) using penta-acetate derivatives

Results
Results and discussion
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