|
Display additional annotations [+]
| Gene |
Clone |
Species |
Stages |
Anatomy |
|
foxj1
|
|
laevis
|
NF stage 10
|
mesoderm
,
dorsal marginal zone
,
superficial
|
|
foxj1
|
|
laevis
|
NF stage 19
|
floor plate
,
neural tube
|
|
foxj1
|
|
laevis
|
NF stage 33 and 34
|
brain
,
hindbrain
,
midbrain-hindbrain boundary
,
spinal cord
,
pronephric nephrostome
,
[+]
|
|
foxj1
|
|
laevis
|
NF stage 45
|
brain
,
spinal cord
,
stomach
,
floor plate
,
filter chamber
,
[+]
|
|
cfap43
|
|
laevis
|
NF stage 10
|
mesoderm
,
dorsal marginal zone
,
superficial
|
|
cfap43
|
|
laevis
|
NF stage 19
|
gastrocoel roof plate
,
floor plate
,
neural tube
|
|
| |
Fig. 2. Expression of cfap43 in wt and foxj1-manipulated Xenopus embryos.
(A) Foxj1 and cfap43 are co-expressed as shown by comparative analysis of foxj1 and cfap43 mRNA expression in staged embryos using specific antisense RNA probes. (a, b) foxj1 expression, but not cfap43 was shown in the precursor of the left-right organizer (LRO), the superficial mesoderm (SM) of stage 10 gastrula embryos. (c, d) Dorsal explants and transversal histological sections (c’, d’) through neurula embryos at stage 19 revealed mRNA transcription of both genes in the floor plate (FP), and cfap43 mRNA in addition in the LRO (GRP; outlined by dashed lines), where expression is induced already at stage 15 (not shown). (e, f) Co-expression in the nephrostomes (black arrowheads) and MCCs (white arrowheads) of stage 33 larvae. (g, h) Staining of stage 45 tadpoles in whole-mounts demonstrated strong neural expression of foxj1 in the sub-commissural organ (SCO), zona limitans intrathalamica (ZLI) and the FP, which was absent for cfap43 (g’’‘, h’’‘). Non-neural expression was found in the dorsal lining of the branchial chamber (BC) (g’‘, h’‘) and in the stomach (stom.) (g’, h’). (B) Cfap43 is a foxj1 target gene. Embryos were unilaterally injected with foxj1 mRNA and analyzed for cfap43 expression. *, injected side. (C) Foxj1 F0 crispant embryos (b, b’) lacked detectable cfap43 expression except for the proctodeum (black arrowhead). Scale bars = 100 μm. |
|
|
| |
cfap43 (cilia and flagella associated protein 43) gene expression in a Xenopus laevis embryo, NF stage 24, (a) lateral view, anterior left, dorsal up, and (a') in high magnification of lateral epidermis. Scale bars = 100 μm. |
|
Display additional annotations [+]
|
| |
Fig. 4. Ciliary defects in Xenopus cfap43 morphant and crispant larval skin.
(A) (a) Kymographs of ciliary motility of single MCCs were generated from control (co) wt, TBMO-, sgRNA1-and sgRNA2-injected specimens. (b) Statistical evaluation of results from 3 independent experiments with 15 embryos each and 5 analyzed MCCs per embryo revealed elevated ciliary beat frequencies upon cfap43 loss-of-function. (B) (a) Schematic depiction of the region used for flow analysis in control wt and manipulated stage 32 embryos. Embryo scheme taken from https://www.xenbase.org/anatomy/alldev.do. (b) Maximum intensity projections of single control wt, TBMO-, sgRNA1-and sgRNA2-injected specimens represent cilia-generated flow. (c) Evaluation of bead transport from 3 independent experiments with 8 analyzed specimens each shows reduced mean velocities in cfap43 morphants and crispants. (C) cfap43 mRNA transcripts were reduced in cfap43 crispant embryos (b, d) at stage 33, with a focus on nephrostomes (a, b) and MCCs (c, d). Morphant Embryos were injected with 0.5 pmol TBMO. (D) IF staining of basal bodies (centrin GFP) and basal feet (Tubg1) in skin MCCs of stage 32 wildtype and crispant specimens. Scale bars: C = 100 μm, D = 10 μm ***, p<0.001. |
|
|
| |
cfap43 (cilia and flagella associated protein 43) gene expression in a Xenopus laevis embryo, NF stage 33 & 34, lateral view, anterior left, dorsal up, showing expression in pronephric nephrostomes. (black arrowheads) and multicilliated epidermal cells (white arrowheads). |
