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CLAPHAM, D.H. and OSTERGREN, G. 1984. Immunocytochemistry of tubulin at meiosis in Tradescantia by a protein-A gold method.–Hereditas 101:137–142. Lund, Sweden. ISSN 0018–0661. A method for the immunocytochemical demonstration of microtubules in pollen mother cells of Tradescantia paludosa is described. Cells were fixed in paraformaldehyde and attached to slides; and cell-walls were partially degraded with enzymes. Then, in the first stage of the indirect staining procedure, monoclonal antibodies to yeast tubulin were applied to the preparations, followed in the second stage by protein-A coupled to colloidal gold. At first metaphase, separately organized cone-shaped bands of microtubules extend from the polar region to each half-bivalent, with the broad end of each cone located near the kinetochore. Most of the microtubules lie between the kinetochores and the nearer pole. A few fibres curve round the half-bivalents and: occasionally cross the equator, but rarely if ever extend into the opposite half-spindle. It appears that kinetochore microtubules are arranged in a manner that does not permit them to interact with fibres from the opposite half-spindle at first metaphase. Further, to judge from the gradient in the density of staining along each cone of microtubules, many or most of the fibres that begin at the kinetochore region do not reach the pole, which reduces the opportunity for kinetochore microtubules to react with polar microtubules in the same half-spindle. |
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