Postnatal Epithelial Growth of the Small Intestine in the Rat Occurs by Both Crypt Fission and Crypt Hyperplasia

Cummins, Adrian G.; Jones, Ben; Thompson, Fiona M.

doi:10.1007/s10620-006-3197-9

Cited by 36 publications

(37 citation statements)

References 31 publications

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“…A major difference in the enteroid system is the high frequency of crypt fission events. We estimate that ϳ5% of crypts freshly isolated from WT murine proximal small intestine show evidence of crypt fission, which is consistent with observations in other species (14,15), whereas multiple crypt fission events in the enteroid cultures are observed during the period 3-8 days postplating. Newly forming crypts are readily recognized as "buds" from existing crypts and therefore can be avoided or specified for study.…”

Section: Discussionsupporting

confidence: 90%

“…In a study validating the putative stem cell marker, leucine-rich repeatcontaining G-protein-coupled receptor 5 (Lgr5), it was shown that freshly isolated small intestinal crypts placed in collagen gels with specific growth factors would form self-organizing crypt organoids (enteroids). Crypts seal at the open end and begin a process whereby new crypts are formed by a process similar to crypt fission in vivo (14,15). The enteroid epithelial cells migrate from the crypt to a central epithelial-lined cavity (villus-like domain) where they are eventually shed in an apoptotic process.…”

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confidence: 99%

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Functional Cftr in crypt epithelium of organotypic enteroid cultures from murine small intestine

Liu

Walker

Cook

et al. 2012

American Journal of Physiology-Cell Physiology

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Physiological studies of intact crypt epithelium have been limited by problems of accessibility in vivo and dedifferentiation in standard primary culture. Investigations of murine intestinal stem cells have recently yielded a primary intestinal culture in three-dimensional gel suspension that recapitulates crypt structure and epithelial differentiation (Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, Van Es JH, Abo A, Kujala P, Peters PJ, Clevers H. Nature 459: 262–265, 2009). We investigated the utility of murine intestinal crypt cultures (termed “enteroids”) for physiological studies of crypt epithelium by focusing on the transport activity of the cystic fibrosis transmembrane conductance regulator Cftr. Enteroids had multiple crypts with well-differentiated goblet and Paneth cells that degranulated on exposure to the muscarinic agonist carbachol. Modified growth medium provided a crypt proliferation rate, as measured by 5-ethynyl-2′-deoxyuridine labeling, which was similar to proliferation in vivo. Immunoblots demonstrated equivalent Cftr expression in comparisons of freshly isolated crypts with primary and passage 1 enteroids. Apparent enteroid differences in mRNA expression of other transporters were primarily associated with villous epithelial contamination of freshly isolated crypts. Microelectrode analysis revealed cAMP-stimulated membrane depolarization in enteroid epithelium from wild-type (WT) but not Cftr knockout (KO) mice. Morphological and microfluorimetric studies, respectively, demonstrated Cftr-dependent cell shrinkage and lower intracellular pH in WT enteroid epithelium in contrast to Cftr KO epithelium or WT epithelium treated with Cftr inhibitor 172. We conclude that crypt epithelium of murine enteroids exhibit Cftr expression and activity that recapitulates crypt epithelium in vivo. Enteroids provide a primary culture model that is suitable for physiological studies of regenerating crypt epithelium.

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Section: Discussionsupporting

confidence: 90%

mentioning

confidence: 99%

Functional Cftr in crypt epithelium of organotypic enteroid cultures from murine small intestine

Liu

Walker

Cook

et al. 2012

American Journal of Physiology-Cell Physiology

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“…1B); pups began to acquire resistance from day 3 and were completely resistant by day 9. Reductions in susceptibility to infection following colonization will be accompanied by structural, physiological, and microbiological changes of GI tract tissues (29,30), and we therefore undertook parallel histological and immunohistochemical investigations of GI tissue in order to ensure that processes that affect susceptibility to infection can be correlated with the dynamic process of postnatal development of the rat intestine. Over the P2-to-P9 period, the length of the digestive system (stomach to colon) increased in an incremental fashion from a mean of 25.6 cm at P2 to 47.6 cm at P9 and was characterized by rapid postnatal physi- ological and anatomical development due to expansion of the small intestine and increased differentiation of the cecum (Fig.…”

Section: Resultsmentioning

confidence: 99%

Altered Innate Defenses in the Neonatal Gastrointestinal Tract in Response to Colonization by Neuropathogenic Escherichia coli

et al. 2013

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d Two-day-old (P2), but not 9-day-old (P9), rat pups are susceptible to systemic infection following gastrointestinal colonization by Escherichia coli K1. Age dependency reflects the capacity of colonizing K1 to translocate from gastrointestinal (GI) tract to blood. A complex GI microbiota developed by P2, showed little variation over P2 to P9, and did not prevent stable K1 colonization. Substantial developmental expression was observed over P2 to P9, including upregulation of genes encoding components of the small intestinal (␣-defensins Defa24 and Defa-rs1) and colonic (trefoil factor Tff2) mucus barrier. K1 colonization modulated expression of these peptides: developmental expression of Tff2 was dysregulated in P2 tissues and was accompanied by a decrease in mucin Muc2. Conversely, ␣-defensin genes were upregulated in P9 tissues. We propose that incomplete development of the mucus barrier during early neonatal life and the capacity of colonizing K1 to interfere with mucus barrier maturation provide opportunities for neuropathogen translocation into the bloodstream. The newborn infant is particularly vulnerable to systemic bacterial infection during the first 4 weeks of life, and mortality and morbidity associated with neonatal bacterial meningitis (NBM) and accompanying sepsis remain significant despite advances in antibacterial chemotherapy and supportive care (1, 2). In the developed world, Escherichia coli and group B streptococci are responsible for the majority of cases of NBM, and bacteria isolated from the cerebrospinal fluid of infected neonates invariably elaborate a protective polysaccharide capsule. Of neuroinvasive E. coli isolates, 80 to 85% express the K1 capsule (3, 4), a homopolymer of ␣-2,8-linked polysialic acid (polySia) that mimics the molecular structure of the polySia modulator of neuronal plasticity in mammalian hosts (5) and enables these strains to evade detection by a neonatal innate immune system undergoing a process of age-dependent maturation (6).Risk factors for NBM include obstetric and perinatal complications, premature birth, and low birth weight, particularly in low socioeconomic groups (7), but predisposition to infection is critically dependent on vertical transmission of the causative agent from mother to infant at or soon after birth (8). Although many aspects of the pathogenesis of E. coli K1 in NBM are unclear, maternally derived E. coli K1 bacteria are known to colonize the neonatal gastrointestinal (GI) tract (8, 9, 10), which is sterile at birth but rapidly acquires a complex microbiota that eventually converges toward a profile characteristic of the adult GI tract (11). E. coli K1 bacteria then translocate from the lumen of the small intestine or colon into the systemic circulation before entering the central nervous system (CNS) across the blood-brain barrier at the cerebral microvascular endothelium of the arachnoid membrane (12) or the blood-cerebrospinal fluid (CSF) barrier at the choroid plexus epithelium (13).Many of the temporal and spatial aspects of NBM can...

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“…This is the case in the intestine in which new crypts are produced by fission of existing ones. This phenomenon likely plays a role in the elongation of the bowel during postnatal growth, but it is not observed in response to injury, suggesting that it might be under control of GH levels, an autonomous program, or both (Cummins et al 2006).…”

Section: Organ Growth Is Driven By Two Distinct Cellular Processesmentioning

confidence: 99%

Organ-Size Regulation in Mammals

Penzo–Méndez

Stanger

2015

Cold Spring Harb Perspect Biol

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The control of organism and organ size is a central question in biology. Despite the attention it has received, our understanding of how adult organ size is determined and maintained is still incomplete. Early work has shown that both autonomous and regulated mechanisms drive vertebrate organ growth, and both intrinsic and extrinsic cues contribute to organ size. The molecular nature of organ-size determinants has been the subject of intense study, and major pathways, which underlie cell interactions controlling cell compartment size,

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Postnatal Epithelial Growth of the Small Intestine in the Rat Occurs by Both Crypt Fission and Crypt Hyperplasia

Cited by 36 publications

References 31 publications

Functional Cftr in crypt epithelium of organotypic enteroid cultures from murine small intestine

Functional Cftr in crypt epithelium of organotypic enteroid cultures from murine small intestine

Altered Innate Defenses in the Neonatal Gastrointestinal Tract in Response to Colonization by Neuropathogenic Escherichia coli

Organ-Size Regulation in Mammals

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