AhR−/− mice retinal cells are functional despite scattered bipolar to RCG connections.

<p>Immunohistochemistry experiments were performed in retina sections of adult AhR+/+ and AhR−/− mice. AhR+/+ and AhR−/− retina at P56 is represented. Retinal ganglion cells (RGC) are labeled with antibodies against ßIII-tubulin; bipolar cells are labeled with antibodies against PKCα (Protein Kinase Cα), nucleus is labeled with DAPI; amacrine and displaced amacrine cells are labeled with both antibodies against calbindin (CaBP)-D28k and calretinin. Horizontal cells are stained with antibodies against calbindin. The density of RGC, bipolar, amacrine and displaced amacrine, and horizontal cells are not significant different between AhR+/+ and AhR−/− mice (see quantifications at the bottom of the figure). The thickness of the ONL is the same between both genotypes. The white arrows in the bottom panel of the PKCa staining indicate a disorganization of the synapses between the bipolar cells and the RGCs in some AhR−/− mice. Scale bar  = 25 µm. ONL: Outer Nuclear Layer, INL: Inner Nuclear Layer, GCL: Ganglion Cells Layer, OPL: Outer Plexiform Layer, IPL: Inner Plexiform Layer.</p

AhR−/−mice have a horizontal pendular nystagmus.

<p>(A) Positions of the eyes of the AhR+/+ mice in both horizontal (EHp) and vertical (EVp) plans in the absence of head movements in the dark. (B) Positions of the eyes of 2 different AhR−/− mice showing spontaneous nystagmus at low frequency in light (upper) and at high frequency in dark (bottom). The AhR−/− mice have an ocular instability exclusively in the horizontal plan whereas the eyes of AhR+/+ mice and AhR+/− mice are stable. (C) Frequencies of the nystagmus in light and dark conditions for each AhR−/− mouse (n = 12). The linear regression is represented by the dotted line in grey. EHp: Eye Horizontal position; EVp: Eye vertical position. In this and all following figures, eye movements to the right are presented upward.</p

Normal vestibular function in AhR−/− mice.

<p>(A) Example of eye movements evoked in AhR+/+ (<i>left</i>) and AhR−/− (<i>right</i>) during sinusoidal oscillation in the horizontal plan at a peak velocity of 50°/s. <i>top</i>: frequency of the oscillation equal to 0.2 Hz; <i>bottom</i>: frequency of 2 Hz. Shaded areas indicate quick phases. At low frequency, the nystagmus overlaps the normal VOR pattern. At high frequency, both AhR+/+ and AhR−/− mice show comparable reflexive eye movements. (B–C), Gain (B) and phase (C) of the VOR in the dark. Blue lines correspond to the AhR+/+ mice, green lines correspond to the AhR+/− mice and red lines correspond to the AhR−/− mice. At low frequency, the VOR gain and phase in the AhR−/− mice decreased compared to the AhR +/+ mice. At high frequency, VOR gain and phase did not differ from that of other groups suggesting that the vestibulo-ocular reflex is not affected in the AhR−/− mice. Asterisk indicates statistical difference with <i>p</i><0.05, EHp, Eye Horizontal position; EVp, Eye Vertical position; Ev, Eye velocity; Tv, Table velocity. In this and all following figures: head velocity traces have been inverted to facilitate comparison with eye velocities; the phase lead is positive (up); plots present mean ± Standard Deviation (SD).</p

The OKR of AhR−/− mice is decreased.

<p>(A) Example of eye movements evoked in both AhR+/+ and AhR−/− mice in horizontal position (EHp) during a rotation at 5°/sec in light in counter-clockwise. (B) OKR gain at 5°/sec. The OKR gain is significantly decreased in the AhR−/− mice compared with AhR+/+ mice. The figure presents discontinuous time, as indicated by the vertical lines. Asterisk indicates statistical difference with p<0.01, EHp: Eye Horizontal position.</p

Electroretinograms are normal in AhR−/− mice.

<p>(A) Representative scotopic ERG at different light intensities (from −2 to −0.49 log cd.s/m²) in both AhR+/+ and AhR−/− mice. Quantifications of a-Wave and b-Wave amplitudes are represented in the lower panel. (B) Representative photopic ERG. The photopic response is obtained using a flash intensity to −0.49 log cd.s/m² on light-adapted mice and measured for both mice. There are no significant differences between both genotypes in scotopic and photopic ERG. Blue lines correspond to the AhR+/+ mice, and red lines correspond to the AhR−/− mice.</p

The AhR is expressed in the eyes during the development of AhR+/+ mice.

<p><i>In situ</i> hybridization is performed on coronal sections of the mice eyes at two embryonic stages (E12 and E14) with digoxigenin-labeled riboprobes for AhR. (A & C) E12 coronal section of AhR+/+ (A) and AhR−/− mice (C); no expression of AhR is detected at this stage. (B & D) E14 coronal section of AhR+/+ (B) and AhR−/− mice (D); the black arrow (in B) indicates the expression of AhR in the retinal ganglion cells. Scale bar represent 250 µm.</p

Cerebellar morphology and function are not affected by the AhR invalidation.

<p>(A) The morphology of the cerebellum in both AhR+/+ (<i>left</i>) and AhR−/−mice (<i>right</i>) is similar. <i>Top:</i> The size of the cerebellum and its foliation are normal in AhR−/−, as shown on sagittal sections labeled with anti-calbindin antibodies and counterstained with DAPI. <i>Bottom:</i> Climbing fibers (stained by VGLUT2) properly innervate the dendritic arborization of Purkinje cells (CaBP). Scale bars represent 250 µm (<i>top</i>) and 25 µm (<i>bottom</i>). (B–C) Normalized gain (C) and phase (D) during the visuo-vestibular conflict at 0.5 Hz. The AhR−/− and AhR+/+ mice are capable of adaptation during and after the vestibulo-ocular conflict. Blue lines correspond to the AhR+/+ mice, and red lines correspond to the AhR−/− mice. Asterisk indicates statistical difference between the VOR gain before and after the conflict with <i>p</i><0.001, # represent the statistical difference between the VOR phase during all the adaptation protocol with p<0.001.</p

Patient Characteristics.

<p>Abbreviations: *Age at diagnosis; ⁺Response to induction therapy, CCR, complete clinical responder; NR, non-responder; <b>^†</b> Alive (A) or Deceased (D) in 2009 ; MA, microarray; TLDA, Taqman low density array card; PF, Cisplatin/5FU; T1PF, Cisplatin/5FU/Paclitaxel; T2PF, Cisplatin/5FU /Docetaxel.</p

Representative HES, p16 and HPV staining of biopsies from NR and CCR individuals.

<p>HES-NR and HES CCR correspond to hematoxylin-eosin-safran staining of representative non-responder and complete clinical responder individuals, respectively, and IHC-p16 and HIS-oncogenic HPV correspond to representative positive immunohistochemistry for the p16 antigen (brown staining) and hybridization <i>in situ</i> for oncogenic HPV DNA (blue punctate nuclear staining), respectively. The bar represents 40 microns in the upper panels and 20 microns in the lower panels.</p

Gene Signature That Predicts Response to PF.

<p>Gene Signature That Predicts Response to PF.</p