Comparative morphology of rodent vestibular periphery. II. Cristae ampullares
Authors: Hussain Ali
Sapan S Desai
A Lysakowski
Anna Lysakowski
Journal: Journal of Neurophysiology (2005)
Discipline: NEUROSCIENCE & BEHAVIOR
Keywords: neuroepithelial preparations; cristae; calretinin immunohistochemistry; calyx afferents; hair cells
Affiliations: Department of Anatomy and Cell Biology, University of Illinois at Chicago, M/C 512 808 S. Wood St., Room 578, Chicago, IL 60612, USA.

Abstract:  We made flattened neuroepithelial preparations of horizontal and vertical (anterior and posterior) cristae from mouse, rat, gerbil, guinea pig, chinchilla, and tree squirrel. Calretinin immunohistochemistry was used to label the calyx class of afferents. Because these afferents are restricted to the central zone of the crista, their distribution allowed us to delineate this zone. In addition to calyx afferents, calretinin also labels approximately 5% of type I hair cells and 20% of type II hair cells throughout the mouse and rat crista epithelium. Measurements of the dimensions of the cristae and counts of hair cells and calyx afferents were determined on all species. Numbers of calyx afferents, hair cells, area, length, and width of the sensory epithelium increase from mouse to tree squirrel. As in the companion paper, we obtained additional data on vestibular end organ dimensions from the literature to construct a power law function describing the relationship between crista surface area and body weight. The vertical cristae of the mouse, rat, and gerbil have an eminentia cruciatum, a region located transversely along the midpoint of the sensory organ and consisting of nonsensory cells. Apart from this eminentia cruciatum, there are no statistical differences between horizontal and vertical cristae with regard to area, width, length, the number and type of hair cells, and number of calretinin-labeled calyx afferents. 

Retracted: No

PubPeer comments:
 Figure 4 appears to be the key figure in this paper. The legend reads  *Horizontal and vertical cristae from the 6 species used in this study are shown.* But I have some concerns about the photos shown in this figure. Many areas in the photos of the cristae appear to contain repetitive features. I marked some of those possible repetitive areas in this figure using boxes of the same color to highlight some of the areas that look unexpectedly similar, without attempting to cover all of them. Could the authors please clarify if there might be some possible biological explanation for these repetitive areas that I could not think of, please? ![file] , 
 Looking at these Melba toast-like images in higher resolution, more repetitive areas appear to become visible. Here is the Chinchilla toast in higher resolution. Boxes of the same color show areas that look similar. ![file] , 
 could be even more duplications in the guinea pig ![file] as well as jpg compression differences in the left part of the tree squirrel ![file] , 
 In the [high-resolution version] , pairs of duplicated features pointed out by Dr Bik can be fused like [Magic Eye] stereoviews; this reveals that many of them fall within much larger, irregular duplicated areas with soft, feathered edges. The  Tree Squirrel PC slice in particular appears to be almost entirely filled with duplications. Note also the uniform feathering around the entire circumference of  Guinea Pig SC : in the center of the slice, even the sub-surface cells are clearly defined, another sign of crude image manipulation. This image also appears in Desai s [thesis](http: search.proquest.com openview 176eb83634d7c4a8cacf81d1cbc5d947 1) (p. 292). , 
 Agree on #4 -first thought that  feathering may be the result of the  enzymatical flattering described in the methods but analysis of the high res pictures shows that feathering was used to mask the composite. ![file] , 
 #4 Here are some of the irregular cloned sections I mentioned. Note that they overlap, so the contents do not appear identical side-by-side, but the identical sections form distinct planes when viewed in stereo. ![file] , 
 Here is the marked up version of Tree Squirrel PC Toast in its high-resolution glory: ![file] , 
 High-resolution redo of the Guinea Pig SC toast. Note that I am reusing colors used in other panels, but that this does not indicate reuse of areas across panels. ![file] , 
 #9 Iu2019m disappointed that the authors have not yet replied, given the numerous concerns. Have any of the previous commentators attempted to communicate with the journal? *** ![file] , 
 High-resolution version of Mouse PC Rat SC Gerbil SC. Note that in this section of Figure 4, there is an overlap between the Rat SC and Gerbil SC panels, in particular on the left side. In addition, the top left part of Rat SC appears to be the same as the bottom right part of the same panel, with a 180 degrees rotation. ![file] , 
 High-resolution version of Mouse HC Rat HC Gerbil HC. There are areas that overlap between Rat SC Gerbil SC Gerbil HC (e.g. ellipses in bright green, dark red, moss, baby blue and mid green), with mirroring or rotations. There are more areas that look repetitive, so please feel free to add more of them. Not that it would further add to the severity of concerns I have about these images. ![file] , 
 https: amp.theguardian.com world 2020 jun 10 surgisphere-sapan-desai-lancet-study-hydroxychloroquine-mass-audit-scientific-papers , 
 I ve been toying with automated methods to determine which tools and parameters work well on known examples of image duplication. The markup below was generated by [sherloq] , which I have to say I m quite impressed with. It s not perfect, but catches areas of duplications between panels and the mirroring within them quite well. ![file] I had to mask the text, as it unsurprisingly generates a ton of noise otherwise. , 
 The University of Illinois in Chicago just let me know that they will not start an investigation because the paper is older than six years. , 
 The journal or the senior author also did not seem to be much concerned, because they issued a Correction in March 2021: https: journals.physiology.org doi full 10.1152 jn.00747.2003_COR  After publication of the article u201cComparative morphology of rodent vestibular periphery. II. Cristae ampullares,u201d it was discovered by an independent postpeer review service (PubPeer) that one figure had alterations that were questioned. The senior author, Anna Lysakowski, has now reexamined all of the original numerical and image data in this and the companion paper u201cComparative morphology of rodent vestibular periphery. I. Saccular and utricular maculaeu201d and has submitted this corrigendum to Fig. 4 in the original article and Fig. 5, which is based on Fig. 4. The flattening procedure described in the methods turned crista ampullares (curved, saddle-shaped structures) into flattened whole mounts and involved both enzymatic and mechanical manipulation, thus some damage was likely inevitable. The corrected figures and figure legends show additional specimens representative of the original flattened images (Fig. 4) and coded calyx (dot) patterns based on these additional specimens (Fig. 5), rather than on the published figure. The corrections impact neither the numerical data nor the overall conclusions of the paper. , 
 Could the senior author perhaps clarify what is meant by the sentences about the sample flattening? Does she imply that the duplications visible in the originally published figure panels are caused by some technical artifact caused by smashing the image or the specimen? I am not satisfied by this explanation. Perhaps some clarification might help. As pointed out by Kaveh Bazargan on Twitter, there is a YouTube video highlighting the duplications, for those who might need a bit more proof: https: www.youtube.com watch?v=WjxngRupl5U ,