Reverse-transcription polymerase chain reaction (RT-PCR) analysis in the development of zebrafish embryos and adult tissues. (A) RT-PCR analysis was performed using RNA extracted from zebrafish embryos 0.5, 6, 12, 14, 24, and 30 hpf, and 2, 3, 4, 7, 14, 21, 28, and 42 dpf. (B) Expression of DPT in various adult tissues. Expression of EF1± served as an internal control. NC, negative control (RNase free water).

In situ hybridization of dermatopontin (DPT) mRNA in adult zebrafish eye. In this and other figures, unless otherwise mentioned, sections were transversely cut with the dorsal on top. Sections (8 μm) were hybridized with the DPT probes. Scale bar = 100 μm. (A) Hybridization of the antisense DPT probe. (B) Hybridization of sense DPT probe. (C) Hematoxylin and eosin staining. Higher magnification images of the green box and red box in (A, B, and C) are shown in (D, G), (E, H), and (F, I), respectively. (J) A schematic diagram showing the process of optic nerve signal transmission. Abbreviations: iri, iris; epi, epithelium; str, stroma; gcl, ganglion cell layer; inl, inner nuclear layer; opl, outer plexiform layer; onl, outer nuclear layer; acl, amacrine cell layer; bcl, bipolar cell layer; pho, photoreceptors.

In situ hybridization of dermatopontin (DPT) mRNA in zebrafish brain. (A, C, E) In situ hybridization with DPT antisense probe. Transverse sections through the telencephalon, mesencephalon, and myelencephalon, respectively. (B, D, and F) Hematoxylin and eosin staining in the brain, counterpart to A, C, and E. (G) In situ hybridization with the DPT sense probe in the myelencephalon. (H) Schematic drawing of the neurogenic zone in the adult zebrafish brain (green areas). Abbreviations: gm, gray matter; ot, optic tectum; nt, neural tract; mo, medulla oblongata; tel, telencephalon; mes, mesencephalon; mye, myelencephalon. Scale bar = 100 μm.

In situ hybridization of DPT mRNA in zebrafish tissues. Hematoxylin and eosin staining in adult zebrafish tissues: (A) skin; (C) spinal cord; (E) dorsal aorta; (G) atrium; (I) ventricle; (K) kidney; (M) gut; (O) notochord; (Q) muscle; (S) fin; (U) bladder. (B, D, F, H, J, L, N, P, R, T, and V) DPT mRNA detection in zebrafish tissues as a counterpart to A, C, E, G, I, K, M, O, Q, S, and U. (W) In situ hybridization of DPT in the regenerating scale; 7 days after scale removal. (X) Kidney section was hybridized with sense DPT probe as a negative control. Abbreviations: mc, Malpighian cell; der, dermis; epi, epidermis; mn, Mauthner neurons; gm, gray matter; hae, hematocyte; myo, myocardium; car, cardiomyocyte; hc, hematopoietic cell; ce, columnar epithelium; vc, vacuolated cell; fis, fibrous inner sheath; eos, elastic outer sheath; sar, sarcolemma; ct, connective tissue; nb, nerve bundle; ep, epithelium; sml, smooth muscle layer; es, episquamal scleroblast; hs, hyposquamal scleroblast; ms, marginal scleroblast. Scale bar = 100 μm.

In situ hybridization of dermatopontin (DPT) mRNA in zebrafish larvae 28 dpf. In these figures, sections were cut frontally from rostral to caudal. The sections (8 µm) were hybridized with the DPT probes. Scale bar is shown bottom left. (A, B) Representative sections of larvae 28 dpf stained using hematoxylin and eosin staining. (C, D) In situ hybridization with DPT antisense probe. DPT mRNA expression was observed in the gut, kidney (C), eye, and skin (D). (E, F) In situ hybridization with DPT sense probes. (G, H) Whole-mount in situ hybridization of the tail zone with DPT antisense probe. (I, J) Whole-mount in situ hybridization of the tail zone with DPT sense probe. Abbreviations: gut, gut; kid, kidney; ski, skin; eye, eye; fin, fin; sca, scale; scale bar = 100 μm.

(A, C, E) Hematoxylin and eosin staining of transverse sections through gill, cartilage, and tooth in adult zebrafish. (B, D, and F) In situ hybridization with DPT antisense probes in counterpart with (A, C, and E). (G and H) In situ hybridization with DPT sense probes in the tooth and gill. Abbreviations: ct, cartilaginous tissue; ep, epithelium; cho, chondrocyte; pc, pulp core. Scale = 100 μm.