CIQUIBIC   05472
CENTRO DE INVESTIGACIONES EN QUIMICA BIOLOGICA DE CORDOBA
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
N O V E L I S O L AT I O N O F C H I C K E N R E T I N A L HORIZONTAL CELLS
Autor/es:
MORERA L P; DÍAZ NM; GUIDO ME.
Lugar:
Córdoba
Reunión:
Jornada; V Jornadas de posgrado; 2011
Institución organizadora:
UNC
Resumen:
We described that the non-visual photopigment melanopsin X is
expressed mainly in horizontal cells of the chicken retina (Verra et
al., 2011). In this work our aim was to purify and culture horizontal
cells (HC´s) from the chicken embryonic retina for further
characterization.
Disaggregated retinas of chicken embryos at day 14 were subjected
to a bovine serum albumin (BSA) discontinuous gradient of
concentrations ranging from 1 to 5%. After centrifugation, cells
collected from the different phases were cultured for 4 days and
characterized by immunochemistry and cell morphology. Phases
were examined with specifics antibodies against HC markers:
PROX-1 and Islet-1. Results showed that only the fraction
corresponding to 2.5% of BSA was highly enriched in PROX-1
positive cells (> 95%) displaying a typical HC morphology. In fact,
some cells in this fraction resembled axon-less candelabrum-shaped
HCs. Preliminary results showed that around 50% of the PROX-1
cells in this phase were positive for Islet-1. Moreover, Western blot
assays demonstrated that mainly the phase at 2.5% BSA, exhibited
positive PROX-1 immunoreactivity (MW: 83kDa).
In conclusion the BSA gradient proved to be the most effective
method to separate different retinal cell populations, and
particularly HC´s which will allow us to characterize them as
potential photoreceptors by biochemical and pharmacological
studies