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The Effect of Bcl‐2 on Apoptosis in the Developing Chick Embryo Brain
Author(s) -
Higdon Lindsay Morgan,
Galileo Deni,
Temburni Murali
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a257-b
Subject(s) - biology , clone (java method) , immunostaining , microbiology and biotechnology , viral replication , virus , programmed cell death , viral vector , apoptosis , virology , gene , immunohistochemistry , immunology , genetics , recombinant dna
During brain development neurons migrate along radial glia to their final destinations. Interactions between radial glial substrates and neuronal integrins facilitate migration. Integrin‐substrate interactions have been shown in other systems to induce expression of Bcl‐2, a protein that suppresses apoptosis (programmed cell death). Bcl‐2 is expressed in early chick optic tectum (midbrain) where it is hypothesized to promote neuronal survival, but its role has not been demonstrated. Thus, a replication‐competent retroviral vector expressing Bcl‐2 was injected into the optic tecta in vivo to increase Bcl‐2 levels and characterize the effects on tectal architecture formation. We have been characterizing Bcl‐2 expression patterns by immunostaining infected and uninfected tectal cryosections. To determine the extent of viral spread we will also immunostain the sections for the viral gag protein. We are also developing an in vitro model that will investigate cell contact mediated survival by using time‐lapse microscopy. We have finished constructing a replication‐incompetent retroviral vector that encodes Bcl‐2 and the marker gene lacZ, which should result in discrete infected cell clones (arrays) with higher numbers of surviving marked neurons. We will infect a group of tecta with this virus and another group with a lacZ‐only expressing virus. We will then count and compare the number of cells per clone produced from each virus type. We predict the number of cells per clone with Bcl‐2‐expressing virus will be greater than control clones. This will implicate Bcl‐2 in mediating cell survival during brain development.