Quantitation of peptide hormone in single cultured secretory neurons of the crab, Cardisoma carnifex.
The content of crustacean hyperglycemic hormone (CHH) in single cultured neurons of the crab Cardisoma carnifex was determined by a sensitive enzyme-linked immunosorbent assay (ELISA), using purified CHH (1-50 pg) of the crab Carcinus maenas as standard. The somata were dissociated from the group of approximately 150 peptidergic neurons that form the X-organ--sinus gland neuroendocrine system. As previously reported, the neurons show immediate regenerative outgrowth in defined culture conditions, and develop, generally, into one of two morphological types: cells that produce broad, lamelliform growth cones (veils), and others that are characterized by branching of neurites. In this study, all but one of 64 veiling cells taken after various times in culture up to 12 days contained CHH. They could be readily categorized as having "high" (> 33 pg; mean 86 +/- 5, S.E., n = 47) or "low" (< or = 33 pg; mean 22 +/- 2.5; n = 17) Carcinus CHH equivalents. Thus, CHH is associated with neurons showing veiling outgrowth, but veiling neurons with low CHH form a distinct, but not morphologically distinguishable group. They may contain an isoform of CHH with limited cross-reactivity. In 24 branching neurons assayed, Carcinus CHH equivalents averaged 7.2 +/- 2 pg. This figure includes 14 neurons in which CHH was undetectable, and one that had 40 pg of Carcinus CHH equivalents. There was no significant change of the hormone content in cells of either type during 6 days of culturing.
Keller
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Békésy Laboratory of Neurobiology, University of Hawaii, Honolulu 96822, USA.
Grau
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S
Cooke
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IM
D000818:Animals; D060829:Arthropod Proteins; D003386:Brachyura; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D007447:Invertebrate Hormones / Q000032:analysis*; D008297:Male; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009474:Neurons / Q000166:cytology*; D009490:Neurosecretory Systems / Q000166:cytology*; D012680:Sensitivity and Specificity
D060829:Arthropod Proteins; D007447:Invertebrate Hormones; D009419:Nerve Tissue Proteins; C015947:hyperglycemic hormone, crustacean