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      Ultrastructural immunolocalization of leptin receptor in mouse brain.

      Neuroendocrinology
      Animals, Brain, ultrastructure, Brain Chemistry, Carrier Proteins, metabolism, Choroid Plexus, Extracellular Space, Hypothalamus, Intracellular Membranes, Mice, Mice, Inbred C57BL, Mice, Obese, Microscopy, Electron, Neuroglia, Receptors, Cell Surface, Receptors, Cytokine, Receptors, Leptin

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          Abstract

          Antibodies directed to amino acids 877-894 (M-18) and 32-51 (K-20) were used to localize leptin receptor by immunocytochemistry in mouse brain. Both antibodies stained several hypothalamic nuclei (paraventricular nucleus, supraoptic nucleus, supraoptic retrochiasmatic nucleus, suprachiasmatic nucleus, preoptic area, ventromedial nucleus, dorsomedial nucleus, lateral hypothalamus, arcuate nucleus, ventral and dorsal premammillary nuclei), the thalamic and amygdaloid nuclei, neurons of the neocortex and archicortex and the epithelial cells of the choroid plexus. While M-18 staining was concentrated in the Golgi area, with K-20 it was dispersed in the cytoplasm. Glial cells were stained only by K-20. These results suggest that the trans-membrane forms of the receptor are concentrated at the membrane level of the Golgi complex of neurons and in epithelial cells of the choroid plexus while the soluble form is dispersed in their cytoplasm. Glial cells express only the soluble form.

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          Identification and expression cloning of a leptin receptor, OB-R.

          The ob gene product, leptin, is an important circulating signal for the regulation of body weight. To identify high affinity leptin-binding sites, we generated a series of leptin-alkaline phosphatase (AP) fusion proteins as well as [125I]leptin. After a binding survey of cell lines and tissues, we identified leptin-binding sites in the mouse choroid plexus. A cDNA expression library was prepared from mouse choroid plexus and screened with a leptin-AP fusion protein to identify a leptin receptor (OB-R). OB-R is a single membrane-spanning receptor most related to the gp130 signal-transducing component of the IL-6 receptor, the G-CSF receptor, and the LIF receptor. OB-R mRNA is expressed not only in choroid plexus, but also in several other tissues, including hypothalamus. Genetic mapping of the gene encoding OB-R shows that it is within the 5.1 cM interval of mouse chromosome 4 that contains the db locus.
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            Abnormal splicing of the leptin receptor in diabetic mice.

            Mutations in the mouse diabetes (db) gene result in obesity and diabetes in a syndrome resembling morbid human obesity. Previous data suggest that the db gene encodes the receptor for the obese (ob) gene product, leptin. A leptin receptor was recently cloned from choroid plexus and shown to map to the same 6-cM interval on mouse chromosome 4 as db. This receptor maps to the same 300-kilobase interval as db, and has at least six alternatively spliced forms. One of these splice variants is expressed at a high level in the hypothalamus, and is abnormally spliced in C57BL/Ks db/db mice. The mutant protein is missing the cytoplasmic region, and is likely to be defective in signal transduction. This suggests that the weight-reducing effects of leptin may be mediated by signal transduction through a leptin receptor in the hypothalamus.
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              Design and synthesis of multi-haem proteins.

              A water-soluble, 62-residue, di-alpha-helical peptide has been synthesized which accommodates two bis-histidyl haem groups. The peptide assembles into a four-helix dimer with 2-fold symmetry and four parallel haems that closely resemble native haems in their spectral and electrochemical properties, including haem-haem redox interaction. This protein is an essential intermediate in the synthesis of molecular 'maquettes', a novel class of simplified versions of the metalloproteins involved in redox catalysis and in energy conversion in respiratory and photosynthetic electron transfer.
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