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Cincinnati Children’s Hospital Medical Center

7 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Studying Wnt Signaling During Patterning of Conducting Airways
John Snowball 1, Manoj Ambalavanan 1, Debora Sinner 1,2
1Neonatology and Pulmonary Biology-Perinatal Institute, Cincinnati Children’s Hospital Medical Center, 2University of Cincinnati, College of Medicine

The use of reporter mice coupled to whole mount and section staining, microscopy and in vivo assays facilitates the analysis of mechanisms underlying the normal patterning of the respiratory tract. Here we describe how these techniques contributed to the analysis of Wnt signaling during tracheal development.

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Developmental Biology

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration
Chandrima Sinha *1,2, Kavisha Arora *1, Anjaparavanda P. Naren 1,2
1Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children’s Hospital Medical Center, 2Department of Physiology, University of Tennessee Health Science Center

MRP4 regulates various cyclic nucleotide-dependent signaling events including a recently elucidated role in cell migration. We describe a direct, but multifaceted approach to unravel the downstream molecular targets of MRP4 resulting in identification of a unique MRP4 interactome that plays key roles in the fine-tuned regulation of fibroblast migration.

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Developmental Biology

Transuterine Fetal Tracheal Occlusion Model in Mice
Emrah Aydın 1,2, Rashika Joshi 3, Marc Oria 1, Foong-Yen Lim 1, Brian Michael Varisco 3, Jose Luis Peiro 1
1The Center for Fetal, Cellular & Molecular Therapy, Cincinnati Fetal Center, Division of Pediatric General and Thoracic Surgery, Cincinnati Children’s Hospital Medical Center, 2Department of Pediatric Surgery, Tekirdag Namik Kemal University School of Medicine, 3The Division of Critical Care Medicine, Department of Pediatrics, Cincinnati Children’s Hospital Medical Center

Various animal models of congenital diaphragmatic hernia and fetal tracheal occlusion present advantages and disadvantages regarding ethical issues, cost, surgical difficulty, size, survival rates, and availability of genetic tools. This model provides a new tool to study the impact of both tracheal occlusion and increased luminal pressure on lung development.

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Developmental Biology

A 3-D Tail Explant Culture to Study Vertebrate Segmentation in Zebrafish
M. Fethullah Simsek 1,3, Ertugrul M. Özbudak 1,2,3
1Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, 2Department of Pediatrics, University of Cincinnati College of Medicine, 3Department of Genetics, Albert Einstein College of Medicine

Here, we present the protocol for 3-D tissue culture of the zebrafish posterior body axis, enabling live study of vertebrate segmentation. This explant model provides control over axis elongation, alteration of morphogen sources, and subcellular resolution tissue-level live imaging.

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Behavior

Using a Murine Model of Psychosocial Stress in Pregnancy as a Translationally Relevant Paradigm for Psychiatric Disorders in Mothers and Infants
Sandra P. Zoubovsky 1,2,3, Akil Wilder 4, Louis Muglia 1,2,3
1Molecular Developmental Biology Program, Cincinnati Children’s Hospital Medical Center, 2Division of Human Genetics, Cincinnati Children’s Hospital Medical Center, 3Department of Pediatrics, University of Cincinnati College of Medicine, 4Division of Veterinary Services, Cincinnati Children’s Hospital Medical Center

The chronic psychosocial stress (CGS) paradigm employs clinically relevant stressors during pregnancy in mice to model psychiatric disorders of mothers and infants. Here, we provide a step-by-step procedure of applying the CGS paradigm and downstream assessments to validate this model.

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Biochemistry

Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes
Daniel Barrera 1, Sierra Upton 2, Megan Rauch 3, Tara Notarianni 4, Ki Suk Eum 5, Megan Liberty 6, Sarmila Venkoba Sah 7, Robert Liu 8, Sean Newcomer 9, Linda E. May 10, Emre Agbas 11, Jessica Sage 12, Edina Kosa 7, Abdulbaki Agbas 7,13
1AdventHelath, 2University of Illinois Chicago School of Public Health, 3Lincoln Memorial University, 4Children’s Mercy Hospital, 5Scripps Clinic, 6Cincinnati Children’s Hospital Medical Center, 7Kansas City University, 8Roblex Rex Veterans Affairs Medical Center, 9California State University San Marcos, 10East Carolina University, 11Massachusetts Institute of Technology, 12Boehringer Ingelheim Norway KS, 13Heartland Center for Mitochondrial Medicine

Preparation of mitochondria-enriched samples from previously frozen archived solid tissues allowed the investigators to perform both functional and analytical assessments of mitochondria in various experimental modalities. This study demonstrates how to prepare mitochondria-enriched preparations from frozen heart tissue and perform analytical assessments of mitochondria.

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Bioengineering

Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality
Gauri Kulkarni *1, Athanasia Apostolou *1, Lorna Ewart 1, Carolina Lucchesi 1, Magdalena Kasendra 2,3
1Emulate Inc. Boston, 2Center for Stem Cell and Organoid Medicine, Cincinnati Children’s Hospital Medical Center, 3Division of Pediatric General and Thoracic Surgery, Cincinnati Children’s Hospital Medical Center

Biopsy-derived intestinal organoids and organ-on-a-chips technologies are combined into a microphysiological platform to recapitulate region-specific intestinal functionality.

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