Intestinal dysmotility in a zebrafish (Danio rerio) shank3a;shank3b mutant model of autism 06 Biological Sciences 0604 Genetics

David M. James, Robert A. Kozol, Yuji Kajiwara, Adam L. Wahl, Emily C. Storrs, Joseph D. Buxbaum, Mason Klein, Baharak Moshiree, Julia Dallman

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background and aims: Autism spectrum disorder (ASD) is currently estimated to affect more than 1% of the world population. For people with ASD, gastrointestinal (GI) distress is a commonly reported but a poorly understood co-occurring symptom. Here, we investigate the physiological basis for GI distress in ASD by studying gut function in a zebrafish model of Phelan-McDermid syndrome (PMS), a condition caused by mutations in the SHANK3 gene. Methods: To generate a zebrafish model of PMS, we used CRISPR/Cas9 to introduce clinically related C-terminal frameshift mutations in shank3a and shank3b zebrafish paralogues (shank3abΔC). Because PMS is caused by SHANK3 haploinsufficiency, we assessed the digestive tract (DT) structure and function in zebrafish shank3abΔC +/- heterozygotes. Human SHANK3 mRNA was then used to rescue DT phenotypes in larval zebrafish. Results: Significantly slower rates of DT peristaltic contractions (p < 0.001) with correspondingly prolonged passage time (p < 0.004) occurred in shank3abΔC +/- mutants. Rescue injections of mRNA encoding the longest human SHANK3 isoform into shank3abΔC +/- mutants produced larvae with intestinal bulb emptying similar to wild type (WT), but still deficits in posterior intestinal motility. Serotonin-positive enteroendocrine cells (EECs) were significantly reduced in both shank3abΔC +/- and shank3abΔC -/- mutants (p < 0.05) while enteric neuron counts and overall structure of the DT epithelium, including goblet cell number, were unaffected in shank3abΔC +/- larvae. Conclusions: Our data and rescue experiments support mutations in SHANK3 as causal for GI transit and motility abnormalities. Reductions in serotonin-positive EECs and serotonin-filled ENS boutons suggest an endocrine/neural component to this dysmotility. This is the first study to date demonstrating DT dysmotility in a zebrafish single gene mutant model of ASD.

Original languageEnglish (US)
Article number3
JournalMolecular Autism
Volume10
Issue number1
DOIs
StatePublished - Jan 31 2019

Fingerprint

Biological Science Disciplines
Zebrafish
Autistic Disorder
Gastrointestinal Tract
Enteroendocrine Cells
Serotonin
Gastrointestinal Motility
Larva
Clustered Regularly Interspaced Short Palindromic Repeats
Gastrointestinal Transit
Haploinsufficiency
Messenger RNA
Frameshift Mutation
Mutation
Goblet Cells
Heterozygote
Genes
Protein Isoforms
Epithelium
Cell Count

Keywords

  • Digestive transit
  • Enteroendocrine
  • Peristaltic rate
  • Phelan-McDermid syndrome

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Neuroscience
  • Developmental Biology
  • Psychiatry and Mental health

Cite this

Intestinal dysmotility in a zebrafish (Danio rerio) shank3a;shank3b mutant model of autism 06 Biological Sciences 0604 Genetics. / James, David M.; Kozol, Robert A.; Kajiwara, Yuji; Wahl, Adam L.; Storrs, Emily C.; Buxbaum, Joseph D.; Klein, Mason; Moshiree, Baharak; Dallman, Julia.

In: Molecular Autism, Vol. 10, No. 1, 3, 31.01.2019.

Research output: Contribution to journalArticle

James, David M. ; Kozol, Robert A. ; Kajiwara, Yuji ; Wahl, Adam L. ; Storrs, Emily C. ; Buxbaum, Joseph D. ; Klein, Mason ; Moshiree, Baharak ; Dallman, Julia. / Intestinal dysmotility in a zebrafish (Danio rerio) shank3a;shank3b mutant model of autism 06 Biological Sciences 0604 Genetics. In: Molecular Autism. 2019 ; Vol. 10, No. 1.
@article{699ad028faa0472f91c894c1db97f1a0,
title = "Intestinal dysmotility in a zebrafish (Danio rerio) shank3a;shank3b mutant model of autism 06 Biological Sciences 0604 Genetics",
abstract = "Background and aims: Autism spectrum disorder (ASD) is currently estimated to affect more than 1{\%} of the world population. For people with ASD, gastrointestinal (GI) distress is a commonly reported but a poorly understood co-occurring symptom. Here, we investigate the physiological basis for GI distress in ASD by studying gut function in a zebrafish model of Phelan-McDermid syndrome (PMS), a condition caused by mutations in the SHANK3 gene. Methods: To generate a zebrafish model of PMS, we used CRISPR/Cas9 to introduce clinically related C-terminal frameshift mutations in shank3a and shank3b zebrafish paralogues (shank3abΔC). Because PMS is caused by SHANK3 haploinsufficiency, we assessed the digestive tract (DT) structure and function in zebrafish shank3abΔC +/- heterozygotes. Human SHANK3 mRNA was then used to rescue DT phenotypes in larval zebrafish. Results: Significantly slower rates of DT peristaltic contractions (p < 0.001) with correspondingly prolonged passage time (p < 0.004) occurred in shank3abΔC +/- mutants. Rescue injections of mRNA encoding the longest human SHANK3 isoform into shank3abΔC +/- mutants produced larvae with intestinal bulb emptying similar to wild type (WT), but still deficits in posterior intestinal motility. Serotonin-positive enteroendocrine cells (EECs) were significantly reduced in both shank3abΔC +/- and shank3abΔC -/- mutants (p < 0.05) while enteric neuron counts and overall structure of the DT epithelium, including goblet cell number, were unaffected in shank3abΔC +/- larvae. Conclusions: Our data and rescue experiments support mutations in SHANK3 as causal for GI transit and motility abnormalities. Reductions in serotonin-positive EECs and serotonin-filled ENS boutons suggest an endocrine/neural component to this dysmotility. This is the first study to date demonstrating DT dysmotility in a zebrafish single gene mutant model of ASD.",
keywords = "Digestive transit, Enteroendocrine, Peristaltic rate, Phelan-McDermid syndrome",
author = "James, {David M.} and Kozol, {Robert A.} and Yuji Kajiwara and Wahl, {Adam L.} and Storrs, {Emily C.} and Buxbaum, {Joseph D.} and Mason Klein and Baharak Moshiree and Julia Dallman",
year = "2019",
month = "1",
day = "31",
doi = "10.1186/s13229-018-0250-4",
language = "English (US)",
volume = "10",
journal = "Acta Veterinaria Scandinavica",
issn = "2040-2392",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Intestinal dysmotility in a zebrafish (Danio rerio) shank3a;shank3b mutant model of autism 06 Biological Sciences 0604 Genetics

AU - James, David M.

AU - Kozol, Robert A.

AU - Kajiwara, Yuji

AU - Wahl, Adam L.

AU - Storrs, Emily C.

AU - Buxbaum, Joseph D.

AU - Klein, Mason

AU - Moshiree, Baharak

AU - Dallman, Julia

PY - 2019/1/31

Y1 - 2019/1/31

N2 - Background and aims: Autism spectrum disorder (ASD) is currently estimated to affect more than 1% of the world population. For people with ASD, gastrointestinal (GI) distress is a commonly reported but a poorly understood co-occurring symptom. Here, we investigate the physiological basis for GI distress in ASD by studying gut function in a zebrafish model of Phelan-McDermid syndrome (PMS), a condition caused by mutations in the SHANK3 gene. Methods: To generate a zebrafish model of PMS, we used CRISPR/Cas9 to introduce clinically related C-terminal frameshift mutations in shank3a and shank3b zebrafish paralogues (shank3abΔC). Because PMS is caused by SHANK3 haploinsufficiency, we assessed the digestive tract (DT) structure and function in zebrafish shank3abΔC +/- heterozygotes. Human SHANK3 mRNA was then used to rescue DT phenotypes in larval zebrafish. Results: Significantly slower rates of DT peristaltic contractions (p < 0.001) with correspondingly prolonged passage time (p < 0.004) occurred in shank3abΔC +/- mutants. Rescue injections of mRNA encoding the longest human SHANK3 isoform into shank3abΔC +/- mutants produced larvae with intestinal bulb emptying similar to wild type (WT), but still deficits in posterior intestinal motility. Serotonin-positive enteroendocrine cells (EECs) were significantly reduced in both shank3abΔC +/- and shank3abΔC -/- mutants (p < 0.05) while enteric neuron counts and overall structure of the DT epithelium, including goblet cell number, were unaffected in shank3abΔC +/- larvae. Conclusions: Our data and rescue experiments support mutations in SHANK3 as causal for GI transit and motility abnormalities. Reductions in serotonin-positive EECs and serotonin-filled ENS boutons suggest an endocrine/neural component to this dysmotility. This is the first study to date demonstrating DT dysmotility in a zebrafish single gene mutant model of ASD.

AB - Background and aims: Autism spectrum disorder (ASD) is currently estimated to affect more than 1% of the world population. For people with ASD, gastrointestinal (GI) distress is a commonly reported but a poorly understood co-occurring symptom. Here, we investigate the physiological basis for GI distress in ASD by studying gut function in a zebrafish model of Phelan-McDermid syndrome (PMS), a condition caused by mutations in the SHANK3 gene. Methods: To generate a zebrafish model of PMS, we used CRISPR/Cas9 to introduce clinically related C-terminal frameshift mutations in shank3a and shank3b zebrafish paralogues (shank3abΔC). Because PMS is caused by SHANK3 haploinsufficiency, we assessed the digestive tract (DT) structure and function in zebrafish shank3abΔC +/- heterozygotes. Human SHANK3 mRNA was then used to rescue DT phenotypes in larval zebrafish. Results: Significantly slower rates of DT peristaltic contractions (p < 0.001) with correspondingly prolonged passage time (p < 0.004) occurred in shank3abΔC +/- mutants. Rescue injections of mRNA encoding the longest human SHANK3 isoform into shank3abΔC +/- mutants produced larvae with intestinal bulb emptying similar to wild type (WT), but still deficits in posterior intestinal motility. Serotonin-positive enteroendocrine cells (EECs) were significantly reduced in both shank3abΔC +/- and shank3abΔC -/- mutants (p < 0.05) while enteric neuron counts and overall structure of the DT epithelium, including goblet cell number, were unaffected in shank3abΔC +/- larvae. Conclusions: Our data and rescue experiments support mutations in SHANK3 as causal for GI transit and motility abnormalities. Reductions in serotonin-positive EECs and serotonin-filled ENS boutons suggest an endocrine/neural component to this dysmotility. This is the first study to date demonstrating DT dysmotility in a zebrafish single gene mutant model of ASD.

KW - Digestive transit

KW - Enteroendocrine

KW - Peristaltic rate

KW - Phelan-McDermid syndrome

UR - http://www.scopus.com/inward/record.url?scp=85060957760&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060957760&partnerID=8YFLogxK

U2 - 10.1186/s13229-018-0250-4

DO - 10.1186/s13229-018-0250-4

M3 - Article

C2 - 30733854

AN - SCOPUS:85060957760

VL - 10

JO - Acta Veterinaria Scandinavica

JF - Acta Veterinaria Scandinavica

SN - 2040-2392

IS - 1

M1 - 3

ER -