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The ultrastructure of isolated glial (Müller) cells from the turtle retina
Abstract:
Sarthy and Lam (1978) described a method for isolation of glial (Müller) cells from turtle retina which involves papain treatment, mechanical dissociation of the retinae, and subsequent separation of the dispersed cells by velocity sedimentation at unit gravity. In order to establish the cytologic integrity and the extent of contamination, we have carried out ultrastructural studies on the isolated cells using scanning and transmission electron microscopy. During the course of these studies we have also examined the effect of papain on the morphology of the retina. Scanning electron microscopic studies show that the isolated cells have a free surface with limited extraneous material attached. The basal portion of the cell is organized as end bulbs with a convoluted surface while the apical region is specialized with long microvilli. Transmission electron microscopy shows that the isolated cells have intact plasma membranes with very few interruptions. The cytoplasm is dense and contains the normal complement of smooth endoplasmic reticulum, microtubules, and filaments. The extent of contamination as estimated from serial sections of a single cell is about 10-15% of the total cell volume. These studies indicate that Müller cells isolated by this technique should be useful for metabolic studies and for characterizing the Müller cell surface.
Insights
Researchers evaluated the cytologic integrity of turtle retinal Müller cells isolated using papain treatment and velocity sedimentation. Ultrastructural studies confirmed the cells
Area of Science:
- Neuroscience
- Cell Biology
- Retinal Research
Background:
- Müller cells are crucial glial cells in the vertebrate retina.
- Previous methods for isolating Müller cells have varying degrees of success.
- Understanding Müller cell integrity is vital for retinal research.
Purpose of the Study:
- To assess the cytologic integrity of turtle retinal Müller cells isolated by Sarthy and Lam (1978).
- To determine the extent of contamination in the isolated Müller cell preparation.
- To evaluate the effect of papain on retinal morphology.
Main Methods:
- Isolation of turtle retinal Müller cells using papain treatment, mechanical dissociation, and velocity sedimentation.
- Ultrastructural analysis via scanning electron microscopy (SEM) and transmission electron microscopy (TEM).
- Examination of retinal tissue morphology following papain treatment.
Main Results:
- Isolated Müller cells exhibited intact plasma membranes and characteristic cellular structures (end bulbs, microvilli).
- SEM revealed a clean free surface on isolated cells.
- TEM indicated minimal contamination (10-15% by cell volume).
- Papain treatment affected retinal morphology.
Conclusions:
- The described isolation method yields turtle retinal Müller cells with high cytologic integrity.
- The isolated cells are suitable for metabolic studies and surface characterization.
- Further investigation into papain's effect on retinal tissue is warranted.