Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
The Cannabinoid Receptor Interacting Proteins 1 of zebrafish are not required for morphological development, viability or fertility.
Fin L
,
Bergamin G
,
Steiner RA
,
Hughes SM
.
???displayArticle.abstract???
The Cannabinoid Receptor Interacting Protein 1 (Cnrip1) was discovered as an interactor with the intracellular region of Cannabinoid Receptor 1 (CB1R, also known as Cnr1 or CB1). Functional assays in mouse show cannabinoid sensitivity changes and Cnrip1 has recently been suggested to control eye development in Xenopus laevis. Two Cnrip1 genes are described in zebrafish, cnrip1a and cnrip1b. In situ mRNA hybridisation revealed accumulation of mRNA encoding each gene primarily in brain and spinal cord, but also elsewhere. For example, cnrip1b is expressed in forming skeletal muscle. CRISPR/Cas9 genome editing generated predicted null mutations in cnrip1a and cnrip1b. Each mutation triggered nonsense-mediated decay of the respective mRNA transcript. No morphological or behavioural phenotype was observed in either mutant. Moreover, fish lacking both Cnrip1a and Cnrip1b both maternally and zygotically are viable and fertile and no phenotype has so far been detected despite strong evolutionary conservation over at least 400 Myr.
Figure 1. Cnrip1 genes in zebrafish. (a) Amino acid sequence alignment of Cnrip1a and Cnrip1b with the two splice variants of the single human CNRIP1 gene, named CRIP1a and CRIP1b. Distinct exons are shown in alternating black and blue font (red indicates amino acids coded by two exons). Comparing both zebrafish genes and CRIP1a asterisk indicates identity and colon similarity between all three, full stop similarity between any two. Bold indicates identity between CRIP1a and CRIP1b. (b) Synteny of adjacent genes in vertebrate CNRIP1 loci. Colours link homologous genes.
Figure 2. Accumulation of mRNAs from cnrip1a and cnrip1b genes. Whole mount in situ mRNA hybridisation of embryos at the indicated stages for antisense probes to cnrip1a (a) and cnrip1b (b). Lateral views (first two images in each panel) are anterior to top dorsal to left, except 24 hpf in which anterior is to left and dorsal to top. Dorsal views (remaining images) are anterior to left flatmounts (a) or wholemount (b), except panel a top right, which is a wholemount montage with anterior to top right. Sense control in inset in panel a top right is anterior to right dorsal to bottom. Quantitative evidence of reproducibility is given in Table S1. tel telencephalon, FB forebrain, MB midbrain, HB hindbrain, SC spinal cord, pf pectoral fin. Bars = 100 µm.
Figure 3. Mutagenesis of cnrip1a and cnrip1b. (a) Alignments of wild type with each of three mutant alleles showing the predicted expressed mutant polypeptide beneath. Bold underline indicates the gRNA target, red font the protospacer motif recognised by Cas9, hyphens deleted bases, blue highlight inserted bases and asterisks novel stop codons. In the mutant protein sequences, bold text indicates the residual wild type fragment and normal font the aberrant polypeptide tail. (b,c). To test for nonsense-mediated decay, about 50 siblings from in-crosses of cnrip1a
+/kg98 (b) and cnrip1b
+/kg101 (c) mutant carriers were subjected to in situ hybridisation for the cognate mRNA at the indicated stages, photographed, DNA extracted, PCR performed across the mutant locus and genotype confirmed by DNA sequencing as indicated beneath each panel. Quantitative evidence of reproducibility is given in Table S1.
Figure 4. Maternal zygotic double cnrip1a
kg98
;cnrip1b
kg101 mutant fish develop normally. (a) Schematic of crosses to generate a cnrip1 double mutant. (b) Darkfield image of 1 dpf double mutant indicating normal complex CNS folds (white arrowhead). (c) Differential interference contrast image of 1 dpf double mutant with arrowheads indicating normal eye (red), ear (purple), notochord (cyan) and haematopoetic tissue (black). (d) Dorsal, lateral and oblique (inset) views of 8 dpf larvae with arrowheads indicating pectoral fins (blue), jaw (turquoise), food traversing gut (green), swim bladder (orange) and xanthophores (yellow). Quantitative evidence of reproducibility is given in Table S1. All fish shown with anterior to left and dorsal up. Bars = 200 μm.
Anavi-Goffer,
Helix 8 Leu in the CB1 cannabinoid receptor contributes to selective signal transduction mechanisms.
2007, Pubmed
Anavi-Goffer,
Helix 8 Leu in the CB1 cannabinoid receptor contributes to selective signal transduction mechanisms.
2007,
Pubmed
Andresen,
Four DNA methylation biomarkers in biliary brush samples accurately identify the presence of cholangiocarcinoma.
2015,
Pubmed
Bethge,
Colorectal cancer DNA methylation marker panel validated with high performance in Non-Hodgkin lymphoma.
2014,
Pubmed
Blume,
Cannabinoid Receptor Interacting Protein 1a Competition with β-Arrestin for CB1 Receptor Binding Sites.
2017,
Pubmed
Blume,
Striatal CB1 and D2 receptors regulate expression of each other, CRIP1A and δ opioid systems.
2013,
Pubmed
Blume,
Cannabinoid receptor interacting protein (CRIP1a) attenuates CB1R signaling in neuronal cells.
2015,
Pubmed
Blume,
Cannabinoid receptor interacting protein suppresses agonist-driven CB1 receptor internalization and regulates receptor replenishment in an agonist-biased manner.
2016,
Pubmed
Boehm,
Exon Junction Complexes: Supervising the Gene Expression Assembly Line.
2016,
Pubmed
Bojnik,
Changes in the cannabinoid (CB1) receptor expression level and G-protein activation in kainic acid induced seizures.
2012,
Pubmed
De Petrocellis,
The endocannabinoid system: a general view and latest additions.
2004,
Pubmed
Di Marzo,
The endocannabinoid system and its therapeutic exploitation.
2004,
Pubmed
Egertová,
Localisation of cannabinoid receptors in the rat brain using antibodies to the intracellular C-terminal tail of CB.
2000,
Pubmed
Elphick,
The evolution and comparative neurobiology of endocannabinoid signalling.
2012,
Pubmed
Fernández-Ruiz,
Endocannabinoids and Neurodegenerative Disorders: Parkinson's Disease, Huntington's Chorea, Alzheimer's Disease, and Others.
2015,
Pubmed
Flores,
Cannabinoid-hypocretin cross-talk in the central nervous system: what we know so far.
2013,
Pubmed
Guggenhuber,
Cannabinoid receptor-interacting protein Crip1a modulates CB1 receptor signaling in mouse hippocampus.
2016,
Pubmed
Hu,
Architecture of cannabinoid signaling in mouse retina.
2010,
Pubmed
Hwang,
Efficient genome editing in zebrafish using a CRISPR-Cas system.
2013,
Pubmed
Jin,
In situ hybridization: methods and applications.
1997,
Pubmed
Koressaar,
Enhancements and modifications of primer design program Primer3.
2007,
Pubmed
Lind,
Identification of an epigenetic biomarker panel with high sensitivity and specificity for colorectal cancer and adenomas.
2011,
Pubmed
Ludányi,
Downregulation of the CB1 cannabinoid receptor and related molecular elements of the endocannabinoid system in epileptic human hippocampus.
2008,
Pubmed
Maccarrone,
Endocannabinoid signaling at the periphery: 50 years after THC.
2015,
Pubmed
Maida,
A user's guide to cannabinoid therapies in oncology.
2016,
Pubmed
Mailleux,
Distribution of cannabinoid receptor messenger RNA in the human brain: an in situ hybridization histochemistry with oligonucleotides.
1992,
Pubmed
Martini,
Ligand-induced down-regulation of the cannabinoid 1 receptor is mediated by the G-protein-coupled receptor-associated sorting protein GASP1.
2007,
Pubmed
Matsuda,
Localization of cannabinoid receptor mRNA in rat brain.
1993,
Pubmed
Matsuda,
Structure of a cannabinoid receptor and functional expression of the cloned cDNA.
1990,
Pubmed
Nie,
Structural domains of the CB1 cannabinoid receptor that contribute to constitutive activity and G-protein sequestration.
2001,
Pubmed
Niehaus,
CB1 cannabinoid receptor activity is modulated by the cannabinoid receptor interacting protein CRIP 1a.
2007,
Pubmed
Oltrabella,
Role of the endocannabinoid system in vertebrates: Emphasis on the zebrafish model.
2017,
Pubmed
Oster,
Identification and validation of highly frequent CpG island hypermethylation in colorectal adenomas and carcinomas.
2011,
Pubmed
Pasquier,
Gene evolution and gene expression after whole genome duplication in fish: the PhyloFish database.
2016,
Pubmed
Pertwee,
Inhibitory effects of certain enantiomeric cannabinoids in the mouse vas deferens and the myenteric plexus preparation of guinea-pig small intestine.
1992,
Pubmed
Piomelli,
The molecular logic of endocannabinoid signalling.
2003,
Pubmed
Sander,
ZiFiT (Zinc Finger Targeter): an updated zinc finger engineering tool.
2010,
Pubmed
Sharma,
Small Molecules from Nature Targeting G-Protein Coupled Cannabinoid Receptors: Potential Leads for Drug Discovery and Development.
2015,
Pubmed
Shaye,
OrthoList: a compendium of C. elegans genes with human orthologs.
2011,
Pubmed
Smith,
Cannabinoid receptor-interacting protein 1a modulates CB1 receptor signaling and regulation.
2015,
Pubmed
Sánchez,
The CB(1) cannabinoid receptor of astrocytes is coupled to sphingomyelin hydrolysis through the adaptor protein fan.
2001,
Pubmed
Thisse,
High-resolution in situ hybridization to whole-mount zebrafish embryos.
2008,
Pubmed
Tiburu,
Human cannabinoid 1 GPCR C-terminal domain interacts with bilayer phospholipids to modulate the structure of its membrane environment.
2011,
Pubmed
Tsou,
Immunohistochemical distribution of cannabinoid CB1 receptors in the rat central nervous system.
1998,
Pubmed
Zheng,
cnrip1 is a regulator of eye and neural development in Xenopus laevis.
2015,
Pubmed
,
Xenbase
