Purpose: To determine the activity, cell specificity, and developmental expression profiles of fragments of the chicken guanylate cyclase activating protein (GCAP)-1 promoter.
Methods: The intrinsic activities of five GCAP1 promoter-luciferase constructs were measured in transiently transfected primary chicken embryonic retinal cultures. Lentivirus vectors carrying GCAP1 promoter-nlacZ transgenes were used to examine the cell specificities and temporal expression characteristics of selected promoter fragments in developing retina.
Results: Three of the five GCAP1 promoter fragments exhibited significant activity in vitro. The expression characteristics of the promoter fragments in vivo varied as a function of promoter length. Expression of nlacZ driven by the 0.6- and 1.7-kb GCAP1 promoter fragments was first observed on embryonic day (E)12 and was restricted to the inner nuclear layer (INL). By E16, nlacZ staining was also detected in the outer nuclear layer (ONL). Expression of nlacZ driven by the 4.2-kb GCAP1 promoter fragment was not observed until E16 and was restricted to the ONL.
Conclusions: The general organization of regulatory cis elements within the GCAP1 promoter is different from other photoreceptor-specific gene promoters. Elements located within 0.3 kb upstream of the transcription start point are capable of producing efficient gene expression; however, additional elements located within 4.0 kb upstream of the transcription start point are necessary to confer on the fragment the cell specificity and developmental expression characteristics of the native GCAP1 promoter. The results of the current study show that the lentiviral vector system is a useful tool for the characterization of the promoters of genes expressed in neural retina.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Retinal guanylyl cyclase activation by calcium sensor proteins mediates photoreceptor degeneration in an mouse model of congenital human blindness.
J Biol Chem
September 2019
Pennsylvania College of Optometry, Salus University, Elkins Park, Pennsylvania 19027.
Deficiency of RD3 (retinal degeneration 3) protein causes recessive blindness and photoreceptor degeneration in humans and in the mouse strain, but the disease mechanism is unclear. Here, we present evidence that RD3 protects photoreceptors from degeneration by competing with guanylyl cyclase-activating proteins (GCAPs), which are calcium sensor proteins for retinal membrane guanylyl cyclase (RetGC). RetGC activity in / retinas was drastically reduced but stimulated by the endogenous GCAPs at low Ca concentrations.
View Article and Find Full Text PDFGene expression networks underlying retinoic acid-induced differentiation of human retinoblastoma cells.
Invest Ophthalmol Vis Sci
March 2003
Mary D Allen Laboratory for Vision Research, Doheny Eye Institute, and Department of Cell and Neurobiology, The Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9112, USA.
Purpose: To understand the genetic regulatory pathways underlying the retinoic acid (RA) induction of cone arrestin, gene array technology and other molecular tools were used to profile global gene expression changes in human retinoblastoma cells.
Methods: Weri-Rb-1 retinoblastoma cells were cultured in the absence or presence of RA for various periods. DNA microarray analysis profiled gene expression followed by real-time PCR and Northern and immunoblot analyses to confirm the change in expression of selected retinal genes and their gene products.
Analyses of the guanylate cyclase activating protein-1 gene promoter in the developing retina.
Invest Ophthalmol Vis Sci
May 2002
Department of Neuroscience, College of Medicine, University of Florida McKnight Brain Institute, 100 Newell Drive, Bldg. 59, Gainesville, FL 32610-0255, USA.
Purpose: To determine the activity, cell specificity, and developmental expression profiles of fragments of the chicken guanylate cyclase activating protein (GCAP)-1 promoter.
Methods: The intrinsic activities of five GCAP1 promoter-luciferase constructs were measured in transiently transfected primary chicken embryonic retinal cultures. Lentivirus vectors carrying GCAP1 promoter-nlacZ transgenes were used to examine the cell specificities and temporal expression characteristics of selected promoter fragments in developing retina.
GCAP1 rescues rod photoreceptor response in GCAP1/GCAP2 knockout mice.
EMBO J
April 2002
Department of Ophthalmology, Moran Eye Center, University of Utah Health Science Center, Salt Lake City, UT 84112-5330, USA.
Visual transduction in retinal photoreceptors operates through a dynamic interplay of two second messengers, Ca(2+) and cGMP. Ca(2+) regulates the activity of guanylate cyclase (GC) and the synthesis of cGMP by acting on a GC-activating protein (GCAP). While this action is critical for rapid termination of the light response, the GCAP responsible has not been identified.
View Article and Find Full Text PDFCharacterization of the chicken GCAP gene array and analyses of GCAP1, GCAP2, and GC1 gene expression in normal and rd chicken pineal.
Mol Vis
July 1999
University of Florida Brain Institute, Department of Neuroscience, Gainesville, FL 32610, USA.
Purpose: This study had three objectives: (1) to characterize the structures of the chicken GCAP1 and GCAP2 genes; (2) to determine if GCAP1, GCAP2, and GC1 genes are expressed in chicken pineal gland; (3) if GC1 is expressed in chicken pineal, to determine if the GC1 null mutation carried by the retinal degeneration (rd) chicken is associated with degenerative changes within the pineal glands of these animals.
Methods: GCAP1 and GCAP2 gene structures were determined by analyses of chicken cosmid and cDNA clones. The putative transcription start points for these genes were determined using 5'-RACE.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!