Analysis Tools
growth/size/body
| N |
• mice are born in approximate Mendelian ratios, overtly normal, and show normal postnatal growth curves up to 24 days of age
|
nervous system
|
• distal connecting cilium (CC) and axonemal compartments are severely perturbed
• at P8, the domain of Ahi1 (a CC compartmental protein) fails to extend distally and become contiguous with Rp1 (a photoreceptor axonemal marker), unlike in most wild-type cilia
• axonemal Rp1 extension is halted after P8, with further shortening of the Ahi1 domain at P14
• quantification of Ahi1 domain length, gap length between Ahi1 and Rp1, and Ahi1 domain length plus gap length shows that the total domain length of Ahi1 + gap is roughly normal, suggesting that the persistent gap of the distal CC is a missing part of the Ahi1 domain
• basal cilia staining of Ift88 (a key intraflagellar transport protein) is attenuated at P8 and P14, with increased intensity at the distal end; a shortened axonemal Ift88 domain is noted at P14 with no major changes in Kif3a staining
• a shortened Rp1 domain length is observed at P8 and P14, whereas the domain length of acetylated alpha-tubulin (spanning the CC) is relatively normal
• however, the array of nine microtubule doublets of the CC is similar to that in wild-type photoreceptors
|
hydrocephaly
(
J:369940
)
|
• all mice exhibit hydrocephalus, as evidenced by enlarged brain ventricles at P56
|
|
• at P56
|
|
• H&E staining indicates enlarged lateral ventricles at P56
|
|
• upon immuno-EM for rhodopsin, many immunogold particles are found at the inner segment (IS) plasma membrane, indicating ectopic localization of rhodopsin
• at P8, PRPH2/RDS and PROM1/CD133 are extensively mislocalized to the IS
|
|
• at P21, the OS disc protein peripherin 2 (PRPH2/RDS) is essentially lost from the OS, with punctate staining in the cell bodies
• starting at P5, rhodopsin localization remains randomly distributed throughout cell bodies rather than being polarized to the ciliary protrusions as in wild-type photoreceptors
• at P8 and P14, OSs appear to be degenerating with reduced levels of rhodopsin and PDE6B
• from P7 to P12, extracellular vesicles of varying sizes accumulate around photoreceptors with occasional intraciliary vesicles
• upon immuno-EM for rhodopsin, disorganized membranous structures are observed in place of OSs, with reduced rhodopsin labeling
• at P8 and P14, extracellular vesicles heavily labeled with rhodopsin are observed
• failed OS morphogenesis appears to be due to aberrant trafficking of rhodopsin and other OS-bound proteins
|
|
• no photoreceptor OSs are found at P8 and P14, when wild-type photoreceptors start to grow OSs distally from the connecting cilium (CC)
• stages S5 and S6 of photoreceptor ciliogenesis, which are specific to photoreceptor OS morphogenesis between P5 and P7, are absent
• however, stages S1 to S4 of photoreceptor ciliogenesis appear normal, with no major changes in the structural features, number of cilia, or length of the CC before P8
|
|
• at P21, rhodopsin (RHO), alpha-transducin (GNAT1), recoverin (RCVRN), and phosphodiesterase 6B (PDE6B) are barely detectable in the rod outer segment (OS); a significant fraction of rhodopsin is mislocalized in the cell bodies
|
|
• at P8 and P14, OSs appear to be degenerating with reduced levels of rhodopsin and PDE6B
|
|
• at P21, cone arrestin is mislocalized throughout the cell bodies
• however, the total number of cones is similar to that in wild-type retinas
|
|
• all mice exhibit rapid retina photoreceptor loss with only one row of nuclei remaining at P42
|
|
• mice exhibit early-onset rod loss
|
reproductive system
azoospermia
(
J:369940
)
 |
|
|
|
• all mice exhibit defective spermatozoa at 6 months of age
|
|
|
• sperm tails are absent at 6 months of age
|
vision/eye
|
• distal connecting cilium (CC) and axonemal compartments are severely perturbed
• at P8, the domain of Ahi1 (a CC compartmental protein) fails to extend distally and become contiguous with Rp1 (a photoreceptor axonemal marker), unlike in most wild-type cilia
• axonemal Rp1 extension is halted after P8, with further shortening of the Ahi1 domain at P14
• quantification of Ahi1 domain length, gap length between Ahi1 and Rp1, and Ahi1 domain length plus gap length shows that the total domain length of Ahi1 + gap is roughly normal, suggesting that the persistent gap of the distal CC is a missing part of the Ahi1 domain
• basal cilia staining of Ift88 (a key intraflagellar transport protein) is attenuated at P8 and P14, with increased intensity at the distal end; a shortened axonemal Ift88 domain is noted at P14 with no major changes in Kif3a staining
• a shortened Rp1 domain length is observed at P8 and P14, whereas the domain length of acetylated alpha-tubulin (spanning the CC) is relatively normal
• however, the array of nine microtubule doublets of the CC is similar to that in wild-type photoreceptors
|
|
• TUNEL staining indicates extensive cell death in the ONL after P14; elevated cell death is detected as early as P14 using anti-Caspase-3 and p-H2A.X antibodies
• however, early developmental cell death in the INL is normal from P5 to P8
|
|
• upon immuno-EM for rhodopsin, many immunogold particles are found at the inner segment (IS) plasma membrane, indicating ectopic localization of rhodopsin
• at P8, PRPH2/RDS and PROM1/CD133 are extensively mislocalized to the IS
|
|
• at P21, the OS disc protein peripherin 2 (PRPH2/RDS) is essentially lost from the OS, with punctate staining in the cell bodies
• starting at P5, rhodopsin localization remains randomly distributed throughout cell bodies rather than being polarized to the ciliary protrusions as in wild-type photoreceptors
• at P8 and P14, OSs appear to be degenerating with reduced levels of rhodopsin and PDE6B
• from P7 to P12, extracellular vesicles of varying sizes accumulate around photoreceptors with occasional intraciliary vesicles
• upon immuno-EM for rhodopsin, disorganized membranous structures are observed in place of OSs, with reduced rhodopsin labeling
• at P8 and P14, extracellular vesicles heavily labeled with rhodopsin are observed
• failed OS morphogenesis appears to be due to aberrant trafficking of rhodopsin and other OS-bound proteins
|
|
• no photoreceptor OSs are found at P8 and P14, when wild-type photoreceptors start to grow OSs distally from the connecting cilium (CC)
• stages S5 and S6 of photoreceptor ciliogenesis, which are specific to photoreceptor OS morphogenesis between P5 and P7, are absent
• however, stages S1 to S4 of photoreceptor ciliogenesis appear normal, with no major changes in the structural features, number of cilia, or length of the CC before P8
|
|
• at P21, rhodopsin (RHO), alpha-transducin (GNAT1), recoverin (RCVRN), and phosphodiesterase 6B (PDE6B) are barely detectable in the rod outer segment (OS); a significant fraction of rhodopsin is mislocalized in the cell bodies
|
|
• at P8 and P14, OSs appear to be degenerating with reduced levels of rhodopsin and PDE6B
|
|
• at P21, cone arrestin is mislocalized throughout the cell bodies
• however, the total number of cones is similar to that in wild-type retinas
|
|
• all mice exhibit rapid retina photoreceptor loss with only one row of nuclei remaining at P42
|
|
• mice exhibit early-onset rod loss
|
|
• OCT imaging shows a much less severe thinning of the inner nuclear layer (INL) at P21 and P42 relative to the observed ONL thinning
|
|
• OCT imaging shows significant thinning of the outer retina encompassing all segments of the photoreceptors as early as P14, with rapid loss of the outer nuclear layer (ONL) in the following weeks
• ONL thickness is reduced to approximately 50% (50 um) by P21, with continued but slower thinning thereafter
|
|
• ONL is completely lost at P42
|
|
• all mice exhibit severe retinal degeneration at P42
|
|
• Muller glia activation in response to retinal injury is noted by P21, as shown by upregulation of glial fibrillary acidic protein
|
|
• at P21, the cone (photopic) response is reduced but still measurable under higher light intensities; by P42, both rod and cone responses are undetectable
• partial preservation of cone ERG function at early postnatal ages likely reflects slower cone loss than rods
|
|
• electroretinography (ERG) shows that the rod (scotopic) response is abolished at P21; by P42, both rod and cone responses are undetectable
|
cellular
|
• distal connecting cilium (CC) and axonemal compartments are severely perturbed
• at P8, the domain of Ahi1 (a CC compartmental protein) fails to extend distally and become contiguous with Rp1 (a photoreceptor axonemal marker), unlike in most wild-type cilia
• axonemal Rp1 extension is halted after P8, with further shortening of the Ahi1 domain at P14
• quantification of Ahi1 domain length, gap length between Ahi1 and Rp1, and Ahi1 domain length plus gap length shows that the total domain length of Ahi1 + gap is roughly normal, suggesting that the persistent gap of the distal CC is a missing part of the Ahi1 domain
• basal cilia staining of Ift88 (a key intraflagellar transport protein) is attenuated at P8 and P14, with increased intensity at the distal end; a shortened axonemal Ift88 domain is noted at P14 with no major changes in Kif3a staining
• a shortened Rp1 domain length is observed at P8 and P14, whereas the domain length of acetylated alpha-tubulin (spanning the CC) is relatively normal
• however, the array of nine microtubule doublets of the CC is similar to that in wild-type photoreceptors
|
azoospermia
(
J:369940
)
|
|
|
|
|
• all mice exhibit defective spermatozoa at 6 months of age
|
|
|
• sperm tails are absent at 6 months of age
|
|
• elevated autophagy in the retina is detected as early as P14 using an anti-LCA3A/B antibody
|
|
• TUNEL staining indicates extensive cell death in the ONL after P14; elevated cell death is detected as early as P14 using anti-Caspase-3 and p-H2A.X antibodies
• however, early developmental cell death in the INL is normal from P5 to P8
|
homeostasis/metabolism
|
• elevated autophagy in the retina is detected as early as P14 using an anti-LCA3A/B antibody
|
