Production and characterization of monoclonal antibodies recognizing head activator in precursor form and immunocytochemical localization of head activator precursor and head activator peptide in the neural cell line NH15-CA2 and in hydra

Differentiation; Research in Biological Diversity
M SchawallerH C Schaller

Abstract

A synthetic gene for the hydra neuropeptide head activator (HA) was used to produce large amounts of an HA bacterial fusion protein. From this protein an HA-containing fragment was cleaved out, attached in high copy number to carrier proteins, and used as an immunogen to produce monoclonal antibodies able to recognize head activator in precursor form. Using such antibodies and others with different specificities for HA epitopes in combination with different fixation procedures, we detected HA immunoreactivity in three locations in the HA-rich neural cell line NH15-CA2. A precursor-like HA immunoreactivity was present in the cytoplasm of cells and detected, independent of fixation procedure, by monoclonal antibodies characterized as HA-precursor-specific. With antibodies specific for the HA peptide, two immunoreactivities could be distinguished, one within cells and one at the outer cell membrane. HA was detected within differentiated cells with long processes when crosslinkers such as carbodiimide or glutaraldehyde were applied together with agents like methanol. HA peptide bound to target cells was restricted to small round cells with an undifferentiated morphology, especially to those in the process of cell division. In hydra...Continue Reading

References

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Citations

Jul 24, 1992·Brain Research. Developmental Brain Research·T T QuachR J Wyatt

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