CIBICI   14215
CENTRO DE INVESTIGACION EN BIOQUIMICA CLINICA E INMUNOLOGIA
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
DEIMINATION AS A POTENTIAL ASTROCYTIC ACTIVATION MARKER FOR RETINAL ASTROCYTES FOLLOWING TEMPERATURE INCUBATION
Autor/es:
ALGECIRAS M; SANJOY K. BHATTACHARYA; HORACIO M. SERRA
Reunión:
Congreso; ARVO 2014; 2014
Resumen:
Purpose: To determine
whether temperature fluctuation elicits an increase in deimination (conversion
of protein bound arginine into citrulline) in retinal astrocytes, and elevated
deimination levels can serve as an astrocytic activation marker (along with
other activation markers) in the retina.
Methods:
Isolated retinal astrocytes obtained from C57BL/6J mice (about 1000 per plate)
were cultured and subjected to different incubation temperatures for one hour,
followed by a stabilization period of 23 hours at 37°C. The exposed and control
astrocytes were evaluated for deimination levels as well as for levels of other
established markers of astrocyte activation (for example, glial fibrillary
acidic protein: GFAP) using immunohistochemical, Western blot and ELISA
analyses.
Results: Optimal
growth of retinal astrocytes occurred at 37°C. Increased levels of astrocyte activation
markers such as Aquaporin4, and Thrombospondin were found in astrocytes
subjected to hypothermia; whereas decreased
GFAP, deimination and peptidylarginine deiminase type 2 (PAD2) levels were
found in cells subjected to this conditions. The astrocytes subjected to
hyperthermia showed an increase in deimination and PAD2 levels. Deimination and
PAD2 levels were higher for hyperthermia (approximately 1.5-fold) than for the
cells subjected to hypothermia. Hyperthermia was accompanied by an increase in
some astrocytic markers such as GFAP and aldehyde dehydrogenase 1 family member
L1 (ALDH1L1).
Conclusion: The
level of deimination undergoes a shift (change in
level compared to baseline) on either side of optimal temperature incubation (37°C)
for retinal astrocytes. The level of deimination correlates with astrocyte activation
markers in the retina when cells are subjected to temperature treatment. These
results agree with our previous work performed on brain cortex astrocytes, also
from the C57BL/6J mouse strain, in which a similar pattern is observed.