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L J Bloem

Publications and source records attributed to L J Bloem.

3 recordsLinked to original sources

The mouse 5-HT1C receptor contains eight hydrophobic domains and is X-linked.

The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) exerts diverse physiological effects in the central and peripheral nervous systems and in smooth muscle by interacting with pharmacologically distinct membrane receptors. We report here the cDNA cloning of the mouse 5-HT1C receptor and its functional expression in Xenopus oocytes. This receptor possesses the unusual feature of containing eight hydrophobic domains capable of forming membrane-spanning alpha-helices, contrary to the usual '7-helix' paradigm for other membrane receptors that function through coupling to GTP-binding proteins. By hybridization analysis of Chinese hamster x mouse somatic cell hybrid lines, the gene for the receptor, designated Htr1c, has been assigned to the mouse X chromosome.

Amino Acid Sequence

Serum stimulation of quiescent human fibroblasts induces the synthesis of tissue factor mRNA followed by the appearance of tissue factor antigen and procoagulant activity.

Expression of the gene for tissue factor, the cell-surface initiator of blood coagulation, was examined by stimulating growth-arrested human fibroblasts with serum and measuring changes in the cellular content of tissue factor mRNA, antigen, and activity. Maximum tissue factor mRNA levels were reached within 1 h following serum induction and slowly declined to basal levels from 24 to 48 h after stimulation. The appearance of the tissue factor mRNA was followed by an increase in tissue factor antigen and activity. The parallel rise in antigenically positive protein and procoagulant activity was first observed about 2 h after serum stimulation with a peak at 12 h followed by a slow decline during the next 36 h. The serum-induced synthesis of the tissue factor mRNA was independent of de novo protein synthesis as demonstrated by the increased tissue factor mRNA levels generated in the presence of cycloheximide. The results of this study suggest that the synthesis of tissue factor in human fibroblasts is regulated principally at the level of transcription. In one strain of fibroblasts the activity/antigen ratio, during the period of maximum synthesis, was indistinguishable from that of tissue factor which had been immunoaffinity purified from human brain and reconstituted into phospholipid vesicles. However, during serum starvation the activity/antigen ratio in these cells was significantly reduced. Western blot analysis revealed that in serum-starved cells there was an accumulation of truncated forms of the tissue factor antigen while in the serum-stimulated cells only the full-length antigen was observed.

Blood