Circadian clocks regulate adenylyl cyclase activity rhythms in human RPE cells

Barbara Pavan, Elena Frigato, Sara Pozzati, Puttur D Prasad, Cristiano Bertolucci, Carla Biondi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Genes and components of the circadian clock may represent relevant drug targets for diseases involving circadian dysfunctions. By exploiting an established cell line derived from human retinal pigment epithelium (HRPE), the cell constituting the blood-retinal barrier that is essential to maintain the visual functions of the sensorineural retina, we showed serum-shock induction of rhythmic changes in forskolin-evoked adenylyl cyclase (AC) activity. In the presence of Ca2+ and protein kinase A, the forskolin-induced AC activity is significantly, but not completely inhibited, suggesting the involvement of both Ca2+-sensitive and Ca2+-insensitive AC isoforms in the regulation of circadian rhythmicity in these cells. Semi-quantitative RT-PCR showed circadian profile in the expression of three AC isoforms, the Ca2+-inhibitable AC5 and AC6 and the Ca2+-insensitive AC7, and the clock genes hPer1 and hPer2. Our results demonstrate for the first time circadian rhythmicity in a human cell line, identifying the isoforms involved in the circadian profile of AC activity and showing a rhythmicity of the clock gene mRNA expression in these cells. Therefore, the results reported here provide evidence for an intertwine between AC/[Ca2+]i signalling pathways and Per genes in the HRPE circadian clockwork.

Original languageEnglish (US)
Pages (from-to)169-173
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume350
Issue number1
DOIs
StatePublished - Nov 10 2006

Fingerprint

Circadian Clocks
Adenylyl Cyclases
Clocks
Periodicity
Genes
Protein Isoforms
Retinal Pigments
Retinal Pigment Epithelium
Colforsin
Cells
Blood-Retinal Barrier
Gene Components
Cell Line
Cyclic AMP-Dependent Protein Kinases
Retina
Shock
Blood
Gene Expression
Polymerase Chain Reaction
Messenger RNA

Keywords

  • Adenylyl cyclase
  • Circadian clock
  • Forskolin
  • HRPE cells
  • PKA
  • Per genes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Circadian clocks regulate adenylyl cyclase activity rhythms in human RPE cells. / Pavan, Barbara; Frigato, Elena; Pozzati, Sara; Prasad, Puttur D; Bertolucci, Cristiano; Biondi, Carla.

In: Biochemical and Biophysical Research Communications, Vol. 350, No. 1, 10.11.2006, p. 169-173.

Research output: Contribution to journalArticle

Pavan, Barbara ; Frigato, Elena ; Pozzati, Sara ; Prasad, Puttur D ; Bertolucci, Cristiano ; Biondi, Carla. / Circadian clocks regulate adenylyl cyclase activity rhythms in human RPE cells. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 350, No. 1. pp. 169-173.
@article{eb45d0f819a24989a9235411ccb8c46b,
title = "Circadian clocks regulate adenylyl cyclase activity rhythms in human RPE cells",
abstract = "Genes and components of the circadian clock may represent relevant drug targets for diseases involving circadian dysfunctions. By exploiting an established cell line derived from human retinal pigment epithelium (HRPE), the cell constituting the blood-retinal barrier that is essential to maintain the visual functions of the sensorineural retina, we showed serum-shock induction of rhythmic changes in forskolin-evoked adenylyl cyclase (AC) activity. In the presence of Ca2+ and protein kinase A, the forskolin-induced AC activity is significantly, but not completely inhibited, suggesting the involvement of both Ca2+-sensitive and Ca2+-insensitive AC isoforms in the regulation of circadian rhythmicity in these cells. Semi-quantitative RT-PCR showed circadian profile in the expression of three AC isoforms, the Ca2+-inhibitable AC5 and AC6 and the Ca2+-insensitive AC7, and the clock genes hPer1 and hPer2. Our results demonstrate for the first time circadian rhythmicity in a human cell line, identifying the isoforms involved in the circadian profile of AC activity and showing a rhythmicity of the clock gene mRNA expression in these cells. Therefore, the results reported here provide evidence for an intertwine between AC/[Ca2+]i signalling pathways and Per genes in the HRPE circadian clockwork.",
keywords = "Adenylyl cyclase, Circadian clock, Forskolin, HRPE cells, PKA, Per genes",
author = "Barbara Pavan and Elena Frigato and Sara Pozzati and Prasad, {Puttur D} and Cristiano Bertolucci and Carla Biondi",
year = "2006",
month = "11",
day = "10",
doi = "10.1016/j.bbrc.2006.09.015",
language = "English (US)",
volume = "350",
pages = "169--173",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Circadian clocks regulate adenylyl cyclase activity rhythms in human RPE cells

AU - Pavan, Barbara

AU - Frigato, Elena

AU - Pozzati, Sara

AU - Prasad, Puttur D

AU - Bertolucci, Cristiano

AU - Biondi, Carla

PY - 2006/11/10

Y1 - 2006/11/10

N2 - Genes and components of the circadian clock may represent relevant drug targets for diseases involving circadian dysfunctions. By exploiting an established cell line derived from human retinal pigment epithelium (HRPE), the cell constituting the blood-retinal barrier that is essential to maintain the visual functions of the sensorineural retina, we showed serum-shock induction of rhythmic changes in forskolin-evoked adenylyl cyclase (AC) activity. In the presence of Ca2+ and protein kinase A, the forskolin-induced AC activity is significantly, but not completely inhibited, suggesting the involvement of both Ca2+-sensitive and Ca2+-insensitive AC isoforms in the regulation of circadian rhythmicity in these cells. Semi-quantitative RT-PCR showed circadian profile in the expression of three AC isoforms, the Ca2+-inhibitable AC5 and AC6 and the Ca2+-insensitive AC7, and the clock genes hPer1 and hPer2. Our results demonstrate for the first time circadian rhythmicity in a human cell line, identifying the isoforms involved in the circadian profile of AC activity and showing a rhythmicity of the clock gene mRNA expression in these cells. Therefore, the results reported here provide evidence for an intertwine between AC/[Ca2+]i signalling pathways and Per genes in the HRPE circadian clockwork.

AB - Genes and components of the circadian clock may represent relevant drug targets for diseases involving circadian dysfunctions. By exploiting an established cell line derived from human retinal pigment epithelium (HRPE), the cell constituting the blood-retinal barrier that is essential to maintain the visual functions of the sensorineural retina, we showed serum-shock induction of rhythmic changes in forskolin-evoked adenylyl cyclase (AC) activity. In the presence of Ca2+ and protein kinase A, the forskolin-induced AC activity is significantly, but not completely inhibited, suggesting the involvement of both Ca2+-sensitive and Ca2+-insensitive AC isoforms in the regulation of circadian rhythmicity in these cells. Semi-quantitative RT-PCR showed circadian profile in the expression of three AC isoforms, the Ca2+-inhibitable AC5 and AC6 and the Ca2+-insensitive AC7, and the clock genes hPer1 and hPer2. Our results demonstrate for the first time circadian rhythmicity in a human cell line, identifying the isoforms involved in the circadian profile of AC activity and showing a rhythmicity of the clock gene mRNA expression in these cells. Therefore, the results reported here provide evidence for an intertwine between AC/[Ca2+]i signalling pathways and Per genes in the HRPE circadian clockwork.

KW - Adenylyl cyclase

KW - Circadian clock

KW - Forskolin

KW - HRPE cells

KW - PKA

KW - Per genes

UR - http://www.scopus.com/inward/record.url?scp=33749251644&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749251644&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2006.09.015

DO - 10.1016/j.bbrc.2006.09.015

M3 - Article

C2 - 16996031

AN - SCOPUS:33749251644

VL - 350

SP - 169

EP - 173

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -