Figure 2.

Identification of CHH in hemolymph and tissue extracts. (A) RIA analysis of fractions from HPLC separation (HPLC RIA) of Sep-Pak purified premolt (stage D₄–E₃₀) hemolymph. The arrow indicates retention time of authentic CHH. (B) HPLC RIA identification of hemolymph extracts from crabs without eyestalks in intermolt (10 ml; stage C₄; n = 5; filled bars) and premolt (13 ml; stage E₅₀; n = 8; open bars). The arrows indicate retention times of authentic CHH and CPRP. (C) Reverse transcription–PCR of total RNA derived from premolt X organs of the eyestalk (XO) and gut tissues. The amplified (421-bp) gut PCR product was cloned and sequenced and was identical to the previously sequenced XO preprohormone cDNA sequence (15).