A Comparison of In Vitro- and In Vivo Phosphorylated Neurofilament Polypeptides

Abstract: Neurofilament polypeptides phosphorylated in vitro by incubation of neurofilament-enriched preparations from rat CNS with [γ-³²P]ATP were compared with the corresponding polypeptides labeled in vivo by injection of ³²P_i into the lateral ventricles of rats. Autoradiography of sodium dodecyl sulfate (SDS)-polyacrylamide gels revealed that the major phosphorylated species in both preparations were the three neurofilament subunits, which have molecular weights of 200K, 145K, and 68K. However, the relative levels of ³²P detected in the three in vitro -labeled subunits differed from the relative in vivo levels. The two larger neurofilament polypeptides displayed similar ³²P isoprotein distribution patterns on two-dimensional gels, whereas additional isoproteins were seen in the in vitro -labeled 68K species. Limited proteolysis in SDS-polyacrylamide gels revealed the presence of common phosphopeptides in the corresponding pairs of in vitro- and in vivo-labeled subunits, but the in vivo -labeled 145K and in vitro -labeled 200K polypeptides contained additional digestion products. Two-dimensional peptide mapping of the 68K polypeptide digested with a mixture of trypsin and chymotrypsin indicated that this component was phosphorylated at a single, identical site, both in vivo and in vitro. These results indicate that the protein kinase that copurifies with neurofilament preparations may be involved in their in vivo phosphorylation.     

A Fossilized Microcenosis In Triassic Amber

ABSTRACT Detailed data on bacterial and protistan microfossils are presented from a 0.003 mm³ piece of Triassic amber (Schlierseerit, Upper Triassic period, 220-230 million years old). This microcenosis, which actually existed as such within a very small, probably semiaquatic habitat, included the remains of about two bacteria species, four fungi ( Palaeodikaryomyces baueri, Pithomyces -like conidia, capillitium-like hyphae, yeast cells) two euglenoids, two chlamydomonas ( Chlamydomonas sp., Chloromonas sp.), two coccal green microalgae ( Chlorella sp., Choricystis -like cells), one zooflagellate, three testate amoebae ( Centropyxis aculeata var. oblonga-like, Cyclopyxis eurystoma -like, Hyalosphenia baueri n. sp.), seven ciliates ( Pseudoplaryophrya nana -like, Mykophagophrys terricola -like, Cyrtolophosis mucicola -like, Paracondylostoma sp., Bryometopus triquetrus -like. Tetrahymena rostrata -like, Paramecium triassicum n. sp.) the microfossils correspond to or diverge from extant species only slightly.     

Vindesine As A Stathmokinetic Agent In Human Rectal Tumours In Organ Culture

Organ culture, using human colorectal mucosa and tumours, is a good system in which to test a new stathmokinetic agent such as vindesine. Using this system we have found that vindesine has similar metaphase-arresting properties to vincristine, including at least a 6-fold dose response difference in its ability to arrest mitosis in mucosa and tumour, mucosa being the more sensitive. Vindesine is a satisfactory stathmokinetic agent, but in view of its greater cost offers no particular advantages over vincristine.