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Nuclei from hypodermis and midgut gland of the crayfish, Orconectes limosus, have been isolated with a slightly modified Chauveau-method. The isolated nuclei are well preserved and free from cytoplasmic contamination. They are able to incorporate [ 3 H]UTP into acid-insoluble material. This labelling is α-amanitin sensitive. The isolated nuclei are devoid of the mitochondrial/microsomal ecdysone-C-20-hydroxylase. The DNA: RNA: protein ratio is 1:0.8: 3.2 (using Triton X-100 during the isolation procedure the ratio is 1:0.4: 4.6) for nuclei from the hypodermis and 1:1: 3.8 for nuclei from the midgut gland. The absolute amount of DNA per cell is 1.6, resp. 3.5 pg. The final DNA-recovery is about 20%. The yield increases to about 60% if the final ultrazentrifugation step is omitted. The isolated nuclei bind  3 H-ecdysone and  3 H-20-OH-ecdysone specifically. As revealed by Scatchard analysis the affinity of the nuclear 20-OH-ecdysone receptor is 1-2 × 10 -9  ᴍ

Isolation of Nuclei from Crayfish Tissues and Demonstration of Nuclear Ecdysteroid Receptors