Tuft Cells: A New Player in Hirschsprung's Disease

Abstract Introduction “Tuft” cells, also known as brush or caveolated cells, are characteristically fusiform shaped, with a distinct apical “tuft” of microvilli extending into the lumen. Double cortin-like kinase 1 (DCLK1) is a microtubule kinase and is a specific marker of intestinal tuft cells. DCLK1-positive tuft cells have been shown to play a key role in gastrointestinal chemosensation, inflammation, and neurotransmission. DCLK1 and Choline acetyltransferase (ChAT), the enzymes responsible for acetylcholine production, are reported to be coexpressed within the gastrointestinal tract. We designed this study to investigate the hypothesis that DCLK1 gene expression is altered in Hirschsprung's disease (HSCR). Materials and Methods HSCR tissue specimens (n = 6) were collected at the time of pull-through surgery, while control samples were obtained at the time of colostomy closure in patients with imperforate anus (n = 6). Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was undertaken to quantify DCLK1 gene expression, and immunolabeling of DCLK1-positive tuft cells was visualized using confocal microscopy. Results qRT-PCR analysis revealed significant downregulation of the DCLK1 gene in both aganglionic and ganglionic HSCR specimens compared with controls (p < 0.05). Confocal microscopy revealed DCLK1-positive tuft cell expression within the colonic mucosa, with a reduction in expression in both aganglionic and ganglionic HSCR colon compared with controls. Conclusion DCLK1 is significantly downregulated in HSCR colon, suggesting a role for tuft cells in cholinergic neurotransmission of the distal colon. The marked decrease in DCLK1 expression within ganglionic specimens highlights the physiologically abnormal nature of this segment in HSCR patients.

Download Full-text

Acquired Aganglionic Megacolon in a Premature Infant: Report of a Case

PEDIATRICS ◽

10.1542/peds.56.3.459 ◽

1975 ◽

Vol 56 (3) ◽

pp. 459-462

Author(s):

Robert J. Touloukian ◽

Raymond Duncan

Keyword(s):

Case Report ◽

Environmental Factors ◽

Premature Infant ◽

Ganglion Cell ◽

Hirschsprung's Disease ◽

Hirschsprung’S Disease ◽

Genetic Factors ◽

Male Infant ◽

Distal Colon ◽

Premature Newborn

Hirschsprung's disease is presumably caused by intrauterine environmental or genetic factors which prevent the migration and formation of the intramural ganglion cell (IMG) in the distal colon. While the IMG is known to be particularly sensitive to anoxemia and other postnatal environmental factors, its selective loss following such stress has not been substantiated in an unoperated patient. The following report of a stressed premature newborn with the clinical and radiographic features of Hirschsprung's disease clearly documents the histologic disappearance of the IMG from the distal colon. CASE REPORT D.J. (#88-65-29), a 1,525-gm male infant, was born to a healthy 22-year-old abortus 0, gravida 1, para 0 mother following an uncomplicated 30-week gestation, ending in a spontaneous uncomplicated delivery.

Download Full-text

Pullthrough Operation for Hirschsprung's Disease: Importance of a Circumferential (Donut) Biopsy at the Level of the Anastomosis

European Journal of Pediatric Surgery Reports ◽

10.1055/s-0039-1693494 ◽

2019 ◽

Vol 07 (01) ◽

pp. e55-e57

Author(s):

Susan Jehangir ◽

Soundappan Venkatraman Sannappa Soundappan ◽

Micheal Krivanek ◽

Susan Arbuckle ◽

Nicole Graf

Keyword(s):

Transition Zone ◽

Transverse Colon ◽

Hirschsprung's Disease ◽

Hirschsprung’S Disease ◽

Ganglion Cells ◽

Distal Colon ◽

Dentate Line ◽

Full Thickness ◽

Mesenteric Border ◽

Antimesenteric Border

AbstractHirschsprung's disease is characterized by the absence of ganglia in the distal colon, resulting in a functional obstruction. It is managed by excision of the aganglionic segment and anastomosis of the ganglionated bowel just above the dentate line. The level of aganglionosis is determined by performing multiple seromuscular biopsies and/or full thickness biopsy on the antimesenteric border of the bowel to determine the level of pullthrough. The transition zone is described as being irregular, and hence a doughnut biopsy is recommended so that the complete circumference can be assessed. Herein, we described a child in whom there was a selective absence of ganglion cells in 30% of the circumference of the bowel along the mesenteric border for most of the transverse colon. This case defies the known concept of neural migration in an intramural and transmesenteric fashion and emphasizes the importance of a doughnut biopsy of the pulled-down segment.

Download Full-text

“Too much guts and not enough brains”: (epi)genetic mechanisms and future therapies of Hirschsprung disease — a review

Clinical Epigenetics ◽

10.1186/s13148-019-0718-x ◽

2019 ◽

Vol 11 (1) ◽

Cited By ~ 6

Author(s):

Emilie G. Jaroy ◽

Lourdes Acosta-Jimenez ◽

Ryo Hotta ◽

Allan M. Goldstein ◽

Ragnhild Emblem ◽

...

Keyword(s):

Gene Expression ◽

Nervous System ◽

Enteric Nervous System ◽

Hirschsprung's Disease ◽

Hirschsprung’S Disease ◽

Current Knowledge ◽

Hirschsprung Disease ◽

Gut Development ◽

Dna And Rna ◽

Expression Of Genes

Abstract Hirschsprung disease is a neurocristopathy, characterized by aganglionosis in the distal bowel. It is caused by failure of the enteric nervous system progenitors to migrate, proliferate, and differentiate in the gut. Development of an enteric nervous system is a tightly regulated process. Both the neural crest cells and the surrounding environment are regulated by different genes, signaling pathways, and morphogens. For this process to be successful, the timing of gene expression is crucial. Hence, alterations in expression of genes specific for the enteric nervous system may contribute to the pathogenesis of Hirschsprung’s disease. Several epigenetic mechanisms contribute to regulate gene expression, such as modifications of DNA and RNA, histone modifications, and microRNAs. Here, we review the current knowledge of epigenetic and epitranscriptomic regulation in the development of the enteric nervous system and its potential significance for the pathogenesis of Hirschsprung’s disease. We also discuss possible future therapies and how targeting epigenetic and epitranscriptomic mechanisms may open new avenues for novel treatment.

Download Full-text

Human Gene Expression Changes Following Diaphragm Unloading Via Controlled Mechanical Ventilation: QRT-PCR Analysis

10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a5038 ◽

2010 ◽

Author(s):

Harsha Deoghare ◽

Tseng-Tien Huang ◽

D Martin

Keyword(s):

Gene Expression ◽

Mechanical Ventilation ◽

Human Gene ◽

Pcr Analysis ◽

Human Gene Expression ◽

Qrt Pcr ◽

Controlled Mechanical Ventilation

Download Full-text

Comparison of Reliable Reference Genes Following Different Hormone Treatments by Various Algorithms for qRT-PCR Analysis of Metasequoia

International Journal of Molecular Sciences ◽

10.3390/ijms20010034 ◽

2018 ◽

Vol 20 (1) ◽

pp. 34 ◽

Cited By ~ 3

Author(s):

Jing-Jing Wang ◽

Shuo Han ◽

Weilun Yin ◽

Xinli Xia ◽

Chao Liu

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Elongation Factor ◽

Pcr Analysis ◽

Qrt Pcr ◽

Gene Normalization ◽

Accurate Normalization ◽

Suitable Reference ◽

Different Tissues ◽

Gene Expression Levels

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most sensitive technique for evaluating gene expression levels. Choosing appropriate reference genes for normalizing target gene expression is important for verifying expression changes. Metasequoia is a high-quality and economically important wood species. However, few systematic studies have examined reference genes in Metasequoia. Here, the expression stability of 14 candidate reference genes in different tissues and following different hormone treatments were analyzed using six algorithms. Candidate reference genes were used to normalize the expression pattern of FLOWERING LOCUS T and pyrabactin resistance-like 8. Analysis using the GrayNorm algorithm showed that ACT2 (Actin 2), HIS (histone superfamily protein H3) and TATA (TATA binding protein) were stably expressed in different tissues. ACT2, EF1α (elongation factor-1 alpha) and HIS were optimal for leaves treated with the flowering induction hormone solution, while Cpn60β (60-kDa chaperonin β-subunit), GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and HIS were the best reference genes for treated buds. EF1α, HIS and TATA were useful reference genes for accurate normalization in abscisic acid-response signaling. Our results emphasize the importance of validating reference genes for qRT-PCR analysis in Metasequoia. To avoid errors, suitable reference genes should be used for different tissues and hormone treatments to increase normalization accuracy. Our study provides a foundation for reference gene normalization when analyzing gene expression in Metasequoia.

Download Full-text

Reference gene selection for qRT-PCR analysis of season- and tissue-specific gene expression profiles in the honey bee Apis mellifera

Scientific Reports ◽

10.1038/s41598-020-70965-4 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ji Hyang Jeon ◽

KyungHwan Moon ◽

YeongHo Kim ◽

Young Ho Kim

Keyword(s):

Gene Expression ◽

Apis Mellifera ◽

Honey Bee ◽

Gene Selection ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Pcr Analysis ◽

Specific Gene ◽

Qrt Pcr ◽

Selection For

Download Full-text

Comparison of Stemness and Gene Expression between Gingiva and Dental Follicles in Children

Stem Cells International ◽

10.1155/2016/8596520 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 1

Author(s):

Chung-Min Kang ◽

Seong-Oh Kim ◽

Mijeong Jeon ◽

Hyung-Jun Choi ◽

Han-Sung Jung ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Pcr Analysis ◽

Expression Levels ◽

Qrt Pcr ◽

Stem Cell Source ◽

Regenerative Dentistry ◽

Induced Pluripotent

The aim of this study was to compare the differential gene expression and stemness in the human gingiva and dental follicles (DFs) according to their biological characteristics. Gingiva (n=9) and DFs (n=9) were collected from 18 children. Comparative gene expression profiles were collected using cDNA microarray. The expression of development, chemotaxis, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSs) related genes was assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Histological analysis was performed using hematoxylin-eosin and immunohistochemical staining. Gingiva had greater expression of genes related to keratinization, ectodermal development, and chemotaxis whereas DFs exhibited higher expression levels of genes related to tooth and embryo development. qRT-PCR analysis showed that the expression levels of iPSc factors includingSOX2,KLF4, andC-MYCwere58.5±26.3,12.4±3.5, and12.2±1.9times higher in gingiva andVCAM1(CD146) andALCAM(CD166) were33.5±6.9and4.3±0.8times higher in DFs. Genes related to MSCs markers includingCD13,CD34,CD73,CD90, andCD105were expressed at higher levels in DFs. The results of qRT-PCR and IHC staining supported the microarray analysis results. Interestingly, this study demonstrated transcription factors of iPS cells were expressed at higher levels in the gingiva. Given the minimal surgical discomfort and simple accessibility, gingiva is a good candidate stem cell source in regenerative dentistry.

Download Full-text