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Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models
Published on: November 21, 2015
Paneth cells in the developing gut: when do they arise and when are they immune competent?
Fardou H Heida1,2, Gaia Beyduz1,2, Marian L C Bulthuis2
1Department of Pediatric Surgery, Beatrix Children's Hospital, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Insights
Paneth cells (PCs) and immune-competent PCs increase with gestational age (GA). Immune-competent PCs emerge by 29 weeks GA, potentially linking to necrotizing enterocolitis (NEC) risk in preterm infants.
Area of Science:
- Perinatal development
- Gastrointestinal immunology
- Developmental biology
Background:
- Paneth cells (PCs) are crucial for intestinal immunity, but their perinatal development and relation to necrotizing enterocolitis (NEC) remain unclear.
- Understanding PC emergence and immune competence during gestation is vital for neonatal health.
Purpose of the Study:
- To determine the gestational timing of Paneth cell (PC) appearance and immune competence.
- To investigate the relationship between PC development and gestational age (GA).
Main Methods:
- Analysis of 57 fetal/infant ileum tissue samples (9-40 weeks GA) via hematoxylin-eosin staining and anti-human defensin 5 immunohistochemistry.
- Semi-quantitative assessment of PC and immune-competent PC numbers per crypt based on GA.
Main Results:
- Both PC and immune-competent PC numbers significantly increased with advancing GA.
- Significantly higher PC numbers were found after 37 weeks GA.
- Immune-competent PCs were notably higher in infants with GA >29 weeks compared to those <29 weeks.
Conclusions:
- The development of immune-competent Paneth cells (PCs) by 29 weeks gestational age (GA) parallels the increased incidence of necrotizing enterocolitis (NEC) in preterm infants.
- This finding suggests a potential role for immune-competent PCs in the pathogenesis of NEC.
Background:
Little is known about the perinatal development of Paneth cells (PCs) during gestation and the relation with necrotizing enterocolitis (NEC). We aimed to investigate when PCs arise and when they become immune competent during gestation.
Methods:
We included 57 samples of ileum tissue of fetuses/infants with a gestional age (GA) between 9 and 40 wk taken as part of a standard autopsy procedure. Hematoxylin-eosin staining and anti-human defensin 5 immunohistochemistry were performed. We performed a semi-quantitative assessment of (immune-competent) PC numbers per 10 crypts per tissue section per GA.
Results:
The number of PCs and the number of immune-competent PCs increased with increasing GA (Spearman's ρ = 0.41, P = 0.002 and ρ = 0.61, P < 0.001, respectively). Whereas significantly higher PC numbers were observed after 37 wk gestation (median 7, range 0-12) compared to preterm infants (median 0, range 0-15; P = 0.002), we counted higher numbers of immune-competent PCs already in infants with GA above 29 wk (median 6, range 0-18) compared to infants with GA under 29 wk (median 2, range 0-9; P < 0.001).
Conclusion:
The significant increase of immune-competent PCs starting from a GA of 29 wk mimics the rise in incidence of NEC during a similar postmenstrual age in preterm infants.
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