Comparative physiology of peripheral clocks: clock genes (CLOCK, PER1, PER2, CRY1 and BMAL 1) and their modulation by light and hormones in fish, amphibians and mammals

• Grant number: 06/03381-1
• Support Opportunities: Research Projects - Thematic Grants
• Start date: August 01, 2007
• End date: July 31, 2012
• Field of knowledge: Biological Sciences - Physiology - Compared Physiology
• Principal Investigator: Ana Maria de Lauro Castrucci
• Grantee: Ana Maria de Lauro Castrucci
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Pesquisadores principais: Maria Aparecida Visconti
• Associated researchers: Mauro Cesar Isoldi
• Associated scholarship(s): 11/04048-2 - Modulatory role and signaling pathways of cortisol on the temporal expression of clock genes in ZEM-2S cell line, BP.MS; 11/02475-0 - Clock gene modulation by glutamate ionotropic receptors, BP.MS; 09/05227-8 - Role of adrenal and ovarian steroids on the entrainment mechanism of the biological clock, BP.PD; + associated scholarships 09/02679-5 - Alpha-MSH effect on gene expression of rhodopsin, tyrosinase, and alpha-MSH receptor, subtype MCR1, in the Mus musculus B16 melanocyte., BP.MS; 09/02673-7 - Endothelin effects on gene expression of melanopsins (Opn4x and Opn4m) and of ETC receptor in Xenopus laevis melanophores, BP.MS; 08/01499-0 - Expression of melatonin receptors (MT1 and MT2) and rhodopsin in B16 melanocytes of Mus musculus, BP.MS - associated scholarships

Abstract

Melanopsin can be expressed, as well as other opsins, in skin pigment cells, to mediate photoresponses. Having in mind that cultured cells may constitute peripheral clocks and respond to visible light, we will analyse: 1. the mechanisms to adjust biological clocks in single cells of teleosts, amphibians and mammals; 2. their regulation by varying photoperiod regimes and hormones; We will investigate: 1. whether ZEM-2S embryonic cell line of Danio rerio, melanophores of Xenopus laevis, and B-16 F10 murine melanoma cells are able to cycle genes such as Per1, independently of light:dark cycle; 2. whether opsin expression is rhythmic, dependent on the integrity of the cellular clock and independent of the light:dark cycle; 3. whether opsin and clock gene expressions may be modified by hormones. These hypotheses will be tested by the quantification of luciferin bioluminescence originated from the activation of luciferase located in Per1 promoter, of mRNA (and of proteins whenever possible) of opsinsand clock genes, under light:dark cycles, or constant darkness, in the presence of increasing hormone (melatonin, endothelin and alfa-MSH) concentrations, for increasing periods of time. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ISOLDI, MAURO CESAR; PROVENCIO, IGNACIO; DE LAURO CASTRUCCI, ANA MARIA. Light modulates the melanophore response to alpha-MSH in Xenopus laevis: An analysis of the signal transduction crosstalk mechanisms involved. General and Comparative Endocrinology, v. 165, n. 1, p. 104-110, JAN 1 2010. (01/02460-1, 06/03381-1)