Characterization and cloning of fasciclin I and fasciclin II glycoproteins in the grasshopper.

Abstract

Monoclonal antibodies were previously used to identify two glycoproteins, called fasciclin I and II (70 and 95 kDa, respectively), which are expressed on different subsets of axon fascicles in the grasshopper (Schistocerca americana) embryo. Here the monoclonal antibodies were used to purify these two membrane-associated glycoproteins for further characterization. Fasciclin II appears to be an integral membrane protein, whereas fasciclin I is an extrinsic membrane protein.The amino acid sequences of the amino terminus and fragments of both proteins were determined. Using synthetic oligonucleotide probes and antibody screening, we isolated genomic and cDNA clones. Partial DNA sequences of these clones indicate that they encode fasciclins I and II.Previous studies gave rise to the labeled pathways hypothesis, which predicts that axon fascicles in the embryonic neuropil are differentially labeled by surface recognition molecules used for growth cone guidance (e.g., see refs. 1-6). To identify candidates for axonal recognition molecules, monoclonal antibodies (mAbs) were generated that recognize surface antigens expressed on subsets of axon fascicles in both grasshopper (Schistocerca americana) (7) and Drosophila (8) embryos.These mAbs were used to characterize three different membrane-associated glycoproteins, called fasciclin I and II in the grasshopper and fasciclin III in Drosophila, which have several features in common. All three proteins (i) are expressed on different subsets of axon fascicles during development, (ii) are regionally expressed on particular portions of embryonic neurons where their axons fasciculate together, (iii) are dynamically expressed duiing axon outgrowth, and (iv) are expressed outside the developing nervous system at other times and places.

Identification of two Drosophila TGF-? family members in the grasshopper Schistocerca americana

“…540,000 phage from a ~gtl 1 cDNA library constructed from S. americana staged at 40% of development (Snow et al 1988) were screened at low stringency (hybridization conditions--30% formamide, 5 × SSC [SSC: 0.15 M NaC1, 0.15 M sodium citrate], 1% SDS, 1% BLOTTO [BLOTTO: 10% Carnation nonfat dry milk] at 42°C; wash conditions--2 × SSC, 0.1% SDS at 50°C). An 850-bp subclone (850PN) from D. melanogaster dpp con-…”

Lineage‐based analysis of the development of the central complex of the drosophila brain

“…The antibody was used to screen a 9-12-hour embryonic Drosophila phage-gt11 cDNA library (Snow et al,1988) that identified two phages containing a 435-bp EcoR1 fragment that did not include the full open reading frame. A radiolabeled probe derived from this fragment was used to screen the cDNA library and identify a large open reading frame (Hortsch et al, 1990).…”

Embryonic development and evolutionary origin of the orthopteran auditory organs

“…Fasciclin I and Fasciclin I1 are membrane-associated glycoproteins that are expressed in an ordered, temporally and spatially dynamic pattern on different neurons in the developing central and peripheral nervous system of the grasshopper and other insects. (Bastiani et al, 1987;Harrelson and Goodman, 1988;Snow, Zinn, Harrelson, et al, 1988;Zinn, McAllister, and Goodman, 1988). The Fasciclins are thought to be specific cell-cell recognition molecules with adhesive-binding properties.…”