Puff development and DNA synthesis in Sciara salivary gland chromosomes in tissue culture

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Abstract
A method for the tissue culture of salivary glands of Sciara coprophila in a chemically defined medium is presented. When the chromosomes of the posterior lobe of the gland have puffs and the chromosomes of the anterior lobe do not have puffs prior to culture, puffs will develop in the anterior lobe chromosomes within a 24 hour culture period. When these chromosome II puffs are at their largest, fully extended state and a bulb on chromosome III is in a larva nearing pupation, the puffs and bulb become condensed in culture more rapidly than in normal development. This condensed state of the chromatin is thought to be heterochromatin. The uptake of tritiated thymidine was compared in homologous glands of 20 larvae in six different stages of development and all glands were labeled before and after culture, indicating that the culture method does not interfere with DNA synthesis.