PUBLICATION

Orphan cytochrome P450 20a1 CRISPR/Cas9 mutants and neurobehavioral phenotypes in zebrafish

Authors

Brun, N.R., Salanga, M.C., Mora-Zamorano, F.X., Lamb, D.C., Goldstone, J.V., Stegeman, J.J.

ID

ZDB-PUB-211216-7

Date

2021

Source

Scientific Reports 11: 23892 (Journal)

Registered Authors

Salanga, Matthew, Stegeman, John J.

Keywords

none

MeSH Terms

Adaptation, Physiological
Loss of Function Mutation
Zebrafish
Anxiety/genetics*
Homozygote
Movement*
CRISPR-Cas Systems
Visual Perception*
Animals

(all 9)

PubMed

34903767 Full text @ Sci. Rep.

Abstract

Orphan cytochrome P450 (CYP) enzymes are those for which biological substrates and function(s) are unknown. Cytochrome P450 20A1 (CYP20A1) is the last human orphan P450 enzyme, and orthologs occur as single genes in every vertebrate genome sequenced to date. The occurrence of high levels of CYP20A1 transcripts in human substantia nigra and hippocampus and abundant maternal transcripts in zebrafish eggs strongly suggest roles both in the brain and during early embryonic development. Patients with chromosome 2 microdeletions including CYP20A1 show hyperactivity and bouts of anxiety, among other conditions. Here, we created zebrafish cyp20a1 mutants using CRISPR/Cas9, providing vertebrate models with which to study the role of CYP20A1 in behavior and other neurodevelopmental functions. The homozygous cyp20a1 null mutants exhibited significant behavioral differences from wild-type zebrafish, both in larval and adult animals. Larval cyp20a1-/- mutants exhibited a strong increase in light-simulated movement (i.e., light-dark assay), which was interpreted as hyperactivity. Further, the larvae exhibited mild hypoactivity during the adaptation period of the optomotor assays. Adult cyp20a1 null fish showed a pronounced delay in adapting to new environments, which is consistent with an anxiety paradigm. Taken together with our earlier morpholino cyp20a1 knockdown results, the results described herein suggest that the orphan CYP20A1 has a neurophysiological role.

Genes / Markers

Figures

Show all Figures

Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
cyp20a1^wh60/wh60 (AB)	control	Adult	swimming behavior decreased process quality, abnormal	Figure 3
cyp20a1^wh60/wh60 (AB)	standard conditions	Day 6	optomotor response decreased process quality, abnormal	Figure 2
cyp20a1^wh60/wh60 (AB)	lighting conditions	Day 6	optomotor response increased process quality, abnormal	Figure 2
cyp20a1^wh60/wh60 (AB)	vibration	Day 6	swimming behavior increased process quality, abnormal	Figure 2
cyp20a1^wh61/wh61 (AB)	control	Adult	swimming behavior decreased process quality, abnormal	Figure 3
cyp20a1^wh61/wh61 (AB)	standard conditions	Day 6	optomotor response decreased process quality, abnormal	Figure 2
cyp20a1^wh61/wh61 (AB)	standard conditions	Day 6	swim bladder uninflated, abnormal	Fig. S1
cyp20a1^wh61/wh61 (AB)	standard conditions	Adult	whole organism decreased pigmentation, abnormal	text only
cyp20a1^wh61/wh61 (AB)	lighting conditions	Day 6	optomotor response increased process quality, abnormal	Figure 2
cyp20a1^wh61/wh61 (AB)	vibration	Day 6	swimming behavior increased process quality, abnormal	Figure 2

1 - 10 of 10

Show

Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
wh60		Indel	cyp20a1
wh61		Complex	cyp20a1

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cyp20a1	CRISPR2-cyp20a1	CRISPR

Fish

Fish
cyp20a1^wh60/wh60 (AB)
cyp20a1^wh61/wh61 (AB)

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Brun et al., 2021 ZDB-PUB-211216-7 PMID:34903767

Orphan cytochrome P450 20a1 CRISPR/Cas9 mutants and neurobehavioral phenotypes in zebrafish

Brun et al., 2021

ZDB-PUB-211216-7
PMID:34903767