Molecular phylogeny of hyperoliid treefrogs: biogeographic origin of Malagasy and Seychellean taxa and re-analysis of familial paraphyly

Preparation of small vertebrates cleared after alcian blue staining of cartilage is facilitated by trypsin digestion. Specimens are fixed in formation, washed, skinned, and eviscerated. After staining in a solution of alcian blue in acetic acid-alcohol for 24-48 hours, they are transferred to water through graded alcohols. Excess alcian blue is removed over a period of up to three weeks by changes every 2-3 days of 1% trypsin in approximately one-third-saturated sodium borate. Bony tissues may be stained after this in a solution of alizarin red S in 0.5% KOH. Specimens are bleached if necessary and dehydrated through graded KOH-glycerine mixtures for storage in glycerine. Since alcohol treatment in addition to formalin fixation does not affect results with this method, it should be useful to researchers who want to study the cartilage or cartilaginous skeletons in museum specimens, which are routinely fixed in formalin and stored in alcohol.

Recent discoveries of fossil vertebrates from the Late Cretaceous of Madagascar include several specimens of a large theropod dinosaur. One specimen includes a nearly complete and exquisitely preserved skull with thickened pneumatic nasals, a median frontal horn, and a dorsal projection on the parietals. The new materials are assigned to the enigmatic theropod group Abelisauridae on the basis of a number of unique features. Fossil remains attributable to abelisaurids are restricted to three Gondwanan landmasses: South America, Madagascar, and the Indian subcontinent. This distribution is consistent with a revised paleogeographic reconstruction that posits prolonged links between these landmasses (via Antarctica), perhaps until late in the Late Cretaceous.