. The Biological bulletin. Biology; Zoology; Marine biology. Posterior midinlestine Posterior spiracle Figure 2. Diagram of endogenous ^-galactosidase activity in the late third-instar larva stained at pH After X-gal staining, /3-galactosidase activity (black) is evident in the anterior and posterior spiracles, distinct segments of both the anterior and posterior midintestine, cellular epi- dermis, lymph glands, and eye-antenna imaginal discs. (Anatomy after Bodenstein, 1965; Tyler, 1994.) In the prepupal stage P4, the amount of foamy-ap- pearing tissue with diffuse stain increased, and s
. The Biological bulletin. Biology; Zoology; Marine biology. Posterior midinlestine Posterior spiracle Figure 2. Diagram of endogenous ^-galactosidase activity in the late third-instar larva stained at pH After X-gal staining, /3-galactosidase activity (black) is evident in the anterior and posterior spiracles, distinct segments of both the anterior and posterior midintestine, cellular epi- dermis, lymph glands, and eye-antenna imaginal discs. (Anatomy after Bodenstein, 1965; Tyler, 1994.) In the prepupal stage P4, the amount of foamy-ap- pearing tissue with diffuse stain increased, and stain was now found throughout the developing intestinal system. The fat body showed light staining. Again, there was no staining in the brain or ventral ganglion. Pupal stages (P5-P15). During metamorphosis, the delicate nature of the remodeling tissue disallowed whole- mount dissections, but X-gal staining of cryostat serial sections revealed dramatic changes in staining intensity and distribution (Figs. 6 and 7). After head eversion, P5, and in stage P6, there was an increase in dark, punctate staining which co-localized with the fat-body tissue of the head capsule, thorax, and ab- domen. By stage P6, dark staining was associated with the first pupal spiracles and trachea. The staining asso- ciated with foamy-appearing tissues was no longer found in the head region but persisted in the developing legs Ring gland Corpus allatum R-othoracic gland Trachea Lymph glands Aorta Gastric caeca. Esopha; Brain hemisphere Subesophageal ganglion ^^^^^ ^^j,^ Figure 3. Diagram of late third-instar larval nervous system and lymph glands. X-gal staining at pH of/3-galactosidase activity (black) is found in three pairs of larval lymph glands flanking the aorta. Light X-gal staining was seen in the lymph glands of some active larvae. Pro- nounced staining of these tissues was apparent in all of the inactive larvae sampled. Anterior to left, dorsal is up. (Anatomy after King, 1970.) Antennal
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