Pre-translational control of the differentiated state: Test for free cytoplasmic Hb-Messenger ribonucleoprotein in chick erythroblasts and chick embryo brain

Summary Non-ribosome-bound 9S RNA was prepared from cytoplasmic ribonucleoprotein of erythropoietic and non-erythropoietic tissues. The RNA was assayed for Hb-mRNA content in a cell-free protein synthesizing system. Free Hb-mRNA was found in the cytoplasm of chick erythroblasts but not in chick embryo brain cytoplasm.

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Zusammenfassung 9S RNS wurde aus cytoplasmischen Ribonucleoprotein-Partikeln von Hühnchen-Erythroblasten sowie Hühnerembryo-Gehirn isoliert. Die RNS wurde auf ihren Gehalt an Hb-Messenger-RNS in einem zellfreien System getestet. Freie Hb-Messenger-RNS wurde nur im Cytoplasma von Erythroblasten gefunden.

     

Multilocus markers for mouse genome analysis: PCR amplification based on single primers of arbitrary nucleotide sequence

Polymerase chain reaction (PCR) based on single primers of arbitrary nucleotide sequence provides a powerful marker system for genome analysis because each primer amplifies multiple products, and cloning, sequencing, and hybridization are not required. We have evaluated this typing system for the mouse by identifying optimal PCR conditions; characterizing effects of GC content, primer length, and multiplexed primers; demonstrating considerable variation among a panel of inbred strains; and establishing linkage for several products. Mg²⁺, primer, template, and annealing conditions were identified that optimized the number and resolution of amplified products. Primers with 40% GC content failed to amplify products readily, primers with 50% GC content resulted in reasonable amplification, and primers with 60% GC content gave the largest number of well-resolved products. Longer primers did not necessarily amplify more products than shorter primers of the same proportional GC content. Multiplexed primers yielded more products than either primer alone and usually revealed novel variants. A strain survey showed that most strains could be readily distinguished with a modest number of primers. Finally, linkage for seven products was established on five chromosomes. These characteristics establish single primer PCR as a powerful method for mouse genome analysis.     

Enrichment for the globin coding region in a chromatin fraction from chick reticulocytes by endonuclease digestion.

A nuclease-sensitive fraction was obtained from chick reticulocyte chromatin by brief digestion with an endonuclease (DNAase II, deoxyribonucleate 3'-oligonucleotidohydrolase, EC 3.1.4.6). The nuclease-sensitive fraction typically contained less than 1% of the chromatin-DNA but about 50% or more of the nascent chromatin-bound RNA. Hybridization of chick globin complementary DNA to the DNA component of the nuclease-sensitive fraction of reticulocyte chromatin indicated a 3--5 fold enrichment for the globin coding region of the chromatin. The control experiment utilizing DNA from a nuclease-sensitive fraction of chick liver chromatin did not show a comparable enrichment for the globin coding region. This suggests that the endonuclease-effected enrichment for the globin coding region in the nuclease-sensitive fraction of reticulocyte chromatin is to some degree specific for structural genes transcribed in reticulocytes.     

Effects of exogenous gonadotropic and steroid hormones on the social behaviour and gonadal maturation of male domestic ducklings Anas platyrhynchos L.

Male domestic ducklings were injected during their first month of life with mammalian gonadotrophins (ovine LH or FSH, HMG) or gonadal steroids (testosterone or oestradiol). LH and testosterone stimulated sexual behaviour while oestradiol inhibited the increase of aggression observed in control birds during the experiment. The mammalian gonadotrophins did not increase plasma testosterone but nevertheless they all stimulated the testis growth. Several hypotheses which could explain this finding (stimulation of spermatogenesis without any apparent effect on testosterone) are discussed and the possibility of a direct action of LH on the sexual behaviour is analysed. Social displays were only moderately stimulated by testosterone and not at all by gonadotrophins. The hormonal controls of these behaviour patterns remains thus largely unknown.     

Immunoenzyme assay of human cartilaginous proteoglycans and their antibodies

An ELISA has been developed to measure human cartilaginous proteoglycans (PG) and their auto-antibodies. The assay is described step by step: successively the nature of the microtiter wells, the concentration of the PG to be coated, the optimal dilution of the antiserum, the length of various incubations and their respective temperature, are described. Standard curve obtained with purified PG is linear in a logit-log representation. The sensitivity of the assay is 2 ng/tube. Finally, biological fluids-sérum and synovial fluid-show a good parallelism with PG.