These results suggest that the non-mammalian type opsin 5 (Opn5L2) functions as a second UV sensor in the photoreceptive organs, while it might function as chemosensor using its direct binding ability to agonist all-in a panel of ten poultry cells by quantitative polymerase chain reaction (PCR). coating (IPL). A weakly immunoreactive bipolar cell is also localized to the INL (arrow), as reported previously (George A et al. (2005) Exp. Attention Res. 81, 616C625).(TIF) pone.0031534.s005.tif (2.3M) GUID:?34BD43DE-A6A5-468F-BF29-98B9DB4ED650 Abstract A mammalian type opsin 5 (neuropsin) is a recently identified ultraviolet (UV)-sensitive pigment of the retina and additional photosensitive organs in parrots. Two additional opsin 5-related molecules have been found in the genomes Peptide M of non-mammalian vertebrates. However, their functions have not been examined as yet. Here, we determine the molecular properties of a second avian opsin 5, cOpn5L2 (chicken opsin 5-like 2), and its localization in the post-hatch chicken. Spectrophotometric analysis and radionucleotide-binding assay have exposed that cOpn5L2 is definitely a UV-sensitive bistable pigment that couples with the Gi subtype of guanine nucleotide-binding protein (G protein). Like a bistable pigment, it also shows the direct binding ability to agonist all-mRNA in the adrenal gland, which is not NCR2 photoreceptive but an endocrine organ, while lower manifestation was found in the brain and retina. In the protein level, cOpn5L2 immunoreactive cells were present in the chromaffin cells of the adrenal gland. In the brain, cOpn5L2 immunoreactive cells were found in the paraventricular and supraoptic nuclei of the anterior hypothalamus, known for photoreceptive deep mind areas. In the retina, cOpn5L2 protein Peptide M was localized to subsets of cells in the ganglion cell coating and the inner nuclear coating. These results suggest that the non-mammalian type opsin 5 (Opn5L2) functions as a second UV sensor in the photoreceptive organs, while it might function as chemosensor using its direct binding ability to agonist all-in a panel of ten chicken cells by quantitative polymerase chain reaction (PCR). Since relatively higher manifestation was observed in the post-hatching chick adrenal glands, mind, and retina (Number 2A), we focused on characterizing Peptide M the is definitely expressed in part of the adrenal glands of the post-hatching chick (Number 2B, C). In contrast, we could not detect mRNA in the post-hatching retina by standard in situ hybridization with digoxigenin-labeled probes (not demonstrated), which shows lower amount of mRNA manifestation than the level of sensitivity of in situ hybridization. We then raised specific antibodies against peptides related to the N-terminal or C-terminal region of cOpn5L2. We found that both antibodies were specific to cOpn5L2, with anti-cOpn5L2 (N) or anti-cOpn5L2 (C) only identifying cOpn5L2-transfected cells, as demonstrated by western blot analysis (Number 2D). We compared the immunoreactivity of the two antibodies and did not detect noticeable variations in their staining pattern (Number 2E, F). Since the anti-cOpn5L2 (C) antibody exhibited stronger immnunoreactivity with lower background than the anti-cOpn5L (N) antibody, we used the cOpn5L2 (C) antibody for subsequent immunostaining experiments. Open in a separate window Number 2 The manifestation pattern of cOpn5L2.Exposed by quantitative PCR (A), in situ hybridization (B, C), western blot analysis (D), and immunohistochemistry (E, F). mRNA level in retina is referred to as 1. shows no staining in the consecutive section (C). mRNA was recognized in a portion of Peptide M the adrenal glands (Number 2B). Immunohistochemical studies also showed that cOpn5L2 protein was localized to a portion of the adrenal glands (Number 3A). The adrenal glands consist of two unique cell lineages; the adrenal cortex, which is derived from the mesoderm, similar to the urogenital system, and generates steroids, and the medulla, which is derived from the neural crest, Peptide M similar to the sympathetic nervous system, and generates catecholamines. However, in avian adrenal glands, the cortical and medullary cells are intermingled throughout the gland, which is different from that of.
