Tissue-specific expression of phospholipase C encoded by the norpA gene of Drosophila melanogaster.
Mutations in the norpA gene of Drosophila melanogaster severely affect the light-evoked photoreceptor potential with strong mutations rendering the fly blind. Molecular cloning of the norpA gene revealed that it encodes phosphatidylinositol-specific phospholipase C, which enzymes play a pivotal role in one of the largest classes of signaling pathways known. We have used Northern analysis, Western blots, phospholipase C activity assays, and immunohistochemical staining of tissues to examine the tissue-specific expression of the norpA gene and found that it is expressed in a variety of tissues besides the eye. Hybridization of norpA cRNA probe to blots of poly(A+) RNA reveals that the gene encodes at least four transcripts: a 7.5-kilobase (kb) transcript that is expressed in eye and 6.5-, 5.5-, and 5.0-kb transcripts that are expressed in adult body or early stages of development. Antiserum generated against the major gene product of norpA recognizes a 130-kDa protein that is abundant in eyes but severely reduced or absent in norpA mutants, a result which is consistent with previous conclusions that the norpA gene encodes an essential component of the visual system. However, the norpA antiserum also recognizes a 130-kDa protein in adult legs, thorax, and male abdomen, but not female abdomen. These localizations are supported by results of phospholipase C activity assays which show that the norpA mutation reduces phospholipase C activity in each of the tissues in which norpA protein can be detected. Furthermore, immunohistochemical staining of tissue sections with the norpA antiserum demonstrates that the norpA protein is abundant in the retina and ocelli and is present to a lesser extent in the brain and thoracic nervous system. Since some of the above mentioned tissues that express norpA (such as thorax, legs, and abdomen) have no known photoreceptor tissue, we conclude that the norpA gene product is also likely to have a role in signaling pathways other than phototransduction.
Zhu
L
L
Department of Biological Sciences, State University of New York, Buffalo 14260.
McKay
R R
RR
Shortridge
R D
RD
D000818:Animals; D015152:Blotting, Northern; D004331:Drosophila melanogaster / Q000201:enzymology*; D005260:Female; D007106:Immune Sera; D007150:Immunohistochemistry; D008297:Male; D009154:Mutation; D010786:Photoreceptor Cells / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D007106:Immune Sera; D012333:RNA, Messenger; D010738:Type C Phospholipases