Gap junction disappearance in astrocytes and leptomeningeal cells as a consequence of protozoan infection.
Trypanosoma cruzi and Toxoplasma gondii are protozoan parasites capable of causing infections of the nervous system. In order to determine effects of infection by these organisms on intercellular communication in the brain, dye coupling and connexin abundance and distribution were examined in leptomeningeal cells and astrocytes infected with T. cruzi or T. gondii. For both cell types infected with either type of protozoan parasite, intercellular diffusion of intracellularly injected Lucifer Yellow was dramatically reduced. Immunocytochemistry with antibodies specific for connexin43 (in astrocytes) or both connexin43 and connexin26 (for leptomeningeal cells) demonstrated that punctate gap junctional staining was much reduced in infected cells, although uninfected neighbors could display normal connexin abundance and distribution. Western blot analyses revealed that connexin43 abundance in both cell types infected with either parasite was similar to that in uninfected cells. Phosphorylation state of connexin43 (inferred from electrophoretic mobility of connexin43 isoforms) was not significantly affected by the infection process. Immunocytochemistry of whole brains from animals acutely infected with either parasite also showed a marked reduction in connexin43 expression. We conclude that infection of both types of brain cells with either protozoan parasite results in a loss of intercellular communication and organized gap junction plaques without affecting expression levels or posttranslational processing of gap junction proteins. Presumably, these changes in gap junction distribution result from altered targeting of the junctional protein to the plasma membrane, and/or from changes in assembly of subunits into functional channels.
Campos de Carvalho
A C
AC
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Roy
C
C
Hertzberg
E L
EL
Tanowitz
H B
HB
Kessler
J A
JA
Weiss
L M
LM
Wittner
M
M
Dermietzel
R
R
Gao
Y
Y
Spray
D C
DC
D000818:Animals; D001099:Arachnoid / Q000166:cytology / Q000469:parasitology; D001253:Astrocytes / Q000737:chemistry / Q000469:parasitology* / Q000648:ultrastructure; D015153:Blotting, Western; D002450:Cell Communication / Q000502:physiology; D002540:Cerebral Cortex / Q000737:chemistry / Q000166:cytology / Q000469:parasitology; D014355:Chagas Disease / Q000503:physiopathology*; D000072259:Connexin 26; D018031:Connexin 43 / Q000032:analysis; D017630:Connexins / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D017629:Gap Junctions / Q000737:chemistry / Q000502:physiology*; D007546:Isoquinolines / Q000493:pharmacokinetics; D010841:Pia Mater / Q000166:cytology / Q000469:parasitology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014124:Toxoplasmosis, Animal / Q000503:physiopathology*
D018031:Connexin 43; D017630:Connexins; D005456:Fluorescent Dyes; D007546:Isoquinolines; D000072259:Connexin 26; C017475:lucifer yellow