ZFIN ID: ZDB-FIG-180620-40
Leacock et al., 2018 - Structure/Function Studies of the α4 Subunit Reveal Evolutionary Loss of a GlyR Subtype Involved in Startle and Escape Responses. Frontiers in molecular neuroscience   11:23 Full text @ Front. Mol. Neurosci.
ADDITIONAL FIGURES
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 8

Morpholino oligonucleotides (SMO1 and SMO2) block splicing of glra4a. (A) Gross morphology of wild-type embryos and those injected with GlyR α4a translation-blocking (TMO) or splice site (SMO1 and SMO2) morpholinos. (B) Schematic of RT-PCR analysis of glra4a morpholinos confirmed the deletion of exon 7 for α4a-SMO1 (lane 3) and the deletion of part of exon 7 for α4a-SMO2 (lane 4). Intact GlyR α4a exon 7–9 PCR products are seen in the control (393 bp, lane 2) and smaller amounts are also observed in lanes 3 and 4. A fragment of around 178 bp is seen in SMO1, which is made up of exon 6 and 8 only. A fragment of 321 bp in SMO2 contains exons 6 and 8 and part of exon 7, which excludes the region that codes for TM2. GlyR α1 exon 7 is present in control and both SMO fish (lower panel). (C,D) DNA sequencing of GlyR α4a cDNAs from SMO1 and SMO2 treated fish reveals that SMO1 results in a 215 bp deletion, resulting in a frameshift and premature stop codon before M1, while SMO2 results in a 72 bp deletion and loss of 24 amino acids, including the majority of the pore-forming M2 domain.

Gene Expression Details No data available
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-glra4a standard conditions Long-pec eye decreased size, abnormal
Long-pec whole organism morphology, normal
WT + MO3-glra4a standard conditions Long-pec eye decreased size, abnormal
Long-pec whole organism morphology, normal
WT + MO4-glra4a standard conditions Long-pec whole organism morphology, normal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front. Mol. Neurosci.