Diversity and development of beetle horns. (A) Diversity in number, size, location, and shape of horn expression between (i) and within (ii) species of Onthophagus. (B and C) Drosophila model of limb formation (B) compared with the development of a thoracic beetle horn from embryo to adult (C). Cuticle is shown in black, epidermis in blue, including schematic expression domains of the proximodistal patterning genes homothorax (hth, yellow), dachshund (dac, green), and Distal-less (Dll, red). Drosophila legs develop from imaginal discs; epidermal invaginations specified during embryonic development, which grow throughout larval development. Patterning takes place while the disc is a 2-dimensional sheet of tissue, and all disc growth occurs while the disc is invaginated into the body interior. In contrast, beetle horns appear not to be specified during embryonic development. Instead, horns grow from the start as 3-dimensional epidermal outbuddings and all growth is confined to the relatively brief prepupal stage and takes place while the primordium is evaginated into the space between the epidermis and larval cuticle. In addition to a rapid prepupal growth phase, horn expression is also affected at times by a drastic pupal remodeling phase (C1 and C2) during the early pupal stage. During this stage pupal horn primordia are either converted into a future adult structure (C1) or resorbed (C2) via programmed cell death. In the later case, expression of Dll, but not hth or dac, is shifted more posteriorly. (D) Position of dac, hth, and Dll within the basic Drosophila limb patterning network (hh, hedgehog; en, engrailed; dpp, decapentaplegic; wg, wingless; EGFR, epidermal growth factor receptor; al, aristaless; b, bar; bab, bric a brac; exd, extradenticle).