A Novel Gene Family Controls Species-Specific Morphological Traits in Hydra
Figure 7
Silencing of oHym301A in H. oligactis Interferes with Tentacle Formation
(A) Depletion of GFP protein in H. vulgaris AEP Ecto-1 line 3, 7, and 14 d after electroporation with GFP dsRNA. Note dark areas of ectodermal epithelium where GFP transcript and protein are completely depleted.
(B) Electroporation of Ecto-1 animals with oHym301A dsRNA has no effect on GFP expression.
(C) RT-PCR shows depletion of oHym301A transcript by RNAi in H. oligactis polyps at various time-points: 0 d, before electroporation and 4–10 d after electroporation; glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used for equilibration.
(D) Disturbed tentacle formation in H. oligactis after electroporation with oHym301A dsRNA. In situ hybridization 7 d after electroporation indicates that abnormalities in tentacle length are strongly correlated with the absence of oHym301A transcripts. Percentage of animals with normal and aberrant tentacle morphology is shown. Absolute number of experimental animals used is shown in brackets. Three independent RNAi experiments were performed.
(E) Time kinetics of tentacle formation in H. oligactis polyps 7 d after electroporation with oHym301A or GFP dsRNA. Mean number of tentacles per polyp at 42, 66 and 130 h after decapitation (two asterisks, p < 0.01 according to ANOVA test).