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Notch Signaling Promotes Intestinal Crypt Fission in the Infant Rat

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Abstract

Background

Growth of the small intestine in the infant rat is promoted by crypt fission and later by increased crypt cell proliferation. Notch signaling could promote crypt fission. Hes-1 is a Notch target gene.

Aim

We assessed the effect of Notch signaling on intestinal crypt fission and on growth of the intestine in the infant rat.

Methods

Hes-1 expression was determined in the small intestine of litters of Hooded Wistar rats aged between 3 and 72 days. Hes-1 RNA expression was measured by quantitative RT-PCR. Four groups of rats (n = 8 or 9) were injected daily, ip, either with vehicle or with the Notch inhibitor DAPT at doses of 3, 10, and 30 mg/kg, from days 9 to 13 of life, and killed on day 14. A microdissection technique was used to measure crypt fission, mitotic count, and apoptotic count. Data were analyzed by ANOVA and by use of Dunnett’s F test.

Results

Hes-1 expression and crypt fission peaked on day 14. DAPT reduced Hes-1 immunostaining in proportion to dose. DAPT reduced villous area to 72 % (p < 0.01), 53 % (p < 0.001), and 38 % (p < 0.001) of control values for 3, 10 and 30 mg/kg doses, respectively, and reduced crypt fission to 53 % (p < 0.001) and 38 % (p < 0.001) of control values, respectively, for 10 and 30 mg/kg doses. Crypt mitotic count was not affected by any DAPT dose. DAPT at 10 and 30 mg/kg significantly increased apoptosis in crypts, by 6.5 and 4.8-fold, respectively.

Conclusions

We conclude that Notch signaling promotes crypt fission and growth of the intestine by maintaining low apoptosis of crypt cells.

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Acknowledgments

Part of this work was submitted by J Woenig as a thesis to the University of Adelaide for a BHealthSci award in 2010. We thank staff of the Animal House, The Queen Elizabeth Hospital, for their assistance. This study was funded by a grant (508024) from the National Health and Medical Research Council of Australia. GSH was supported by The Sally Birch Cancer Council Australia Senior Research Fellowship in Cancer Control.

Conflicts of interest

None.

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, The Queen Elizabeth Hospital, Woodville South, SA, 5011, Australia

    Adrian G. Cummins, Joshua A. Woenig, Rino P. Donato & Phulwinder K. Grover

  2. Discipline of Medicine, School of Medicine, Faculty of Health Sciences, University of Adelaide, North Terrace, Adelaide, SA, 5000, Australia

    Simon J. Proctor

  3. School of Animal and Veterinary Sciences, The University of Adelaide, Roseworthy Campus, Roseworthy, SA, 5371, Australia

    Gordon S. Howarth

Authors
  1. Adrian G. Cummins
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  2. Joshua A. Woenig
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  3. Rino P. Donato
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  4. Simon J. Proctor
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  6. Phulwinder K. Grover
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Correspondence to Adrian G. Cummins.

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Cummins, A.G., Woenig, J.A., Donato, R.P. et al. Notch Signaling Promotes Intestinal Crypt Fission in the Infant Rat. Dig Dis Sci 58, 678–685 (2013). https://doi.org/10.1007/s10620-012-2422-y

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  • DOI: https://doi.org/10.1007/s10620-012-2422-y

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