Cincinnati Children’s Hospital Medical Center

7 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Studying Wnt Signaling During Patterning of Conducting Airways

John Snowball¹, Manoj Ambalavanan¹, 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.

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.

Developmental Biology

Transuterine Fetal Tracheal Occlusion Model in Mice

Emrah Aydın^1,2, Rashika Joshi³, Marc Oria¹, Foong-Yen Lim¹, Brian Michael Varisco³, Jose Luis Peiro¹

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.

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.

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 Wilder⁴, 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.

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 Barrera¹, Sierra Upton², Megan Rauch³, Tara Notarianni⁴, Ki Suk Eum⁵, Megan Liberty⁶, Sarmila Venkoba Sah⁷, Robert Liu⁸, Sean Newcomer⁹, Linda E. May¹⁰, Emre Agbas¹¹, Jessica Sage¹², Edina Kosa⁷, 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.

Bioengineering

Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality

Gauri Kulkarni^*1, Athanasia Apostolou^*1, Lorna Ewart¹, Carolina Lucchesi¹, 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.