Cell Recognition and Synapse Formation

Nirenberg, M.

Abstract

Lambdagt11 cDNA libraries derived from human brain poly A+ RNA were screened for recombinants that code for Alpha-subunits of G signal transduction proteins. Eleven alpha s and two alpha i clones were characterized. Four species of alpha s cDNA were found. A mechanism for generating the four species of alpha s mRNA by alternative splicing of precursor RNA was proposed. Treatment of NG108-15 neuroblastoma-glioma hybrid cells cAMP for several days results in the appearance of voltage-sensitive calcium channels and other ions channels. Twenty cDNA clones were obtained that hybridize to species of poly A+ RNA that increase in abundance 10-90 fold due to treatment of cells with dibutyryl cAMP. Affinity-purified antibodies to the Alpha or Gamma protein subunits of voltage-sensitive calcium channels were used to screen a Lambdagt11 cDNA library. Twenty putative voltage-sensitive calcium channel Alpha subunit cDNA clones and 29 putative Gamma subunit clones were found. Antigenic molecules termed TOP, which are distributed in a dorsal to ventral concentration gradient in chicken retina, were purified. TOP was shown to be a protein with an Mr of 47,000. The gradient of TOP in the retina is formed early in embryonic development. Thereafter, perpetuation of the gradient does not depend on the continuous presence of an extracellular gradient of diffusable molecules or on maintenance of interactions between cells. Synapses and neurites in the retina of developing chick embryos were reduced markedly by injection of anti-TOP antibody into the eye.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Heart, Lung, and Blood Institute (NHLBI)
• Type: Intramural Research (Z01)
• Project #: 1Z01HL000009-12
• Application #: 3966495
• Support Year: 12
• Fiscal Year: 1986

Institution

• Name: U.S. National Heart Lung and Blood Inst
• Country: United States

Publications

Xia, Menghang; Huang, Ruili; Guo, Vicky et al. (2009) Identification of compounds that potentiate CREB signaling as possible enhancers of long-term memory. Proc Natl Acad Sci U S A 106:2412-7

Koizumi, Keita; Higashida, Haruhiro; Yoo, Siuk et al. (2007) RNA interference screen to identify genes required for Drosophila embryonic nervous system development. Proc Natl Acad Sci U S A 104:5626-31

Sharma, Shail K; Nirenberg, Marshall (2007) Silencing of genes in cultured Drosophila neurons by RNA interference. Proc Natl Acad Sci U S A 104:12925-30

Chen, Xiaobing; Vinade, Lucia; Leapman, Richard D et al. (2005) Mass of the postsynaptic density and enumeration of three key molecules. Proc Natl Acad Sci U S A 102:11551-6

Ivanov, Andrej I; Rovescalli, Alessandra C; Pozzi, Paola et al. (2004) Genes required for Drosophila nervous system development identified by RNA interference. Proc Natl Acad Sci U S A 101:16216-21

Lum, Lawrence; Yao, Shenqin; Mozer, Brian et al. (2003) Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells. Science 299:2039-45

Nirenberg, Marshall (2003) The genetic revolution: the importance of flies and worms. Am J Psychiatry 160:615

Koizumi, Keita; Lintas, Carla; Nirenberg, Marshall et al. (2003) Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: transgenic alterations and tertiary structure. Proc Natl Acad Sci U S A 100:3119-24

Hwang, Kae-Jung; Xiang, Bosong; Gruschus, James M et al. (2003) Distortion of the three-dimensional structure of the vnd/NK-2 homeodomain bound to DNA induced by an embryonically lethal A35T point mutation. Biochemistry 42:12522-31

Wang, Lan-Hsiang; Chmelik, Rebecca; Nirenberg, Marshall (2002) Sequence-specific DNA binding by the vnd/NK-2 homeodomain of Drosophila. Proc Natl Acad Sci U S A 99:12721-6