Hisaka Jingu

Current perspectives on the role of thyroid hormone in growth and development of cerebellum

The thyroid hormone (TH) is essential for growth and development of brain, including the cerebellum. Deficiency of TH during the perinatal period results in abnormal cerebellar development, which is well documented in rodent animal models. TH exerts its major effect by binding to the nuclear TH receptor (TR), a ligand-regulated transcription factor. Although TR is highly expressed in many brain regions, including the cerebellum, TH-target genes that likely play critical roles in brain development have not yet been fully clarified. At present, however, expression of many cerebellar genes is known to be altered by perinatal hypothyroidism. Interestingly, after the critical period of TH action (first 2 weeks of postnatal life in rodent cerebellum), the activities of many genes that are altered by perinatal hypothyroidism return to the same levels as those of euthyroid animal despite morphological alterations. Several prominent candidate genes that may play key roles in TH-mediated cerebellar development are discussed in this review. On the other hand, TR-mediated transcription may be modulated by various substances. The nuclear hormone receptor superfamily contains more than 40 transcriptional factors and, most of these receptors are present in the brain. Possible interactions between TR and such transcription factors are also discussed. Further, several additional issues that need to be clarified are discussed. One such issue is the discrepancy of phenotypes among TR-knockout and perinatal hypothyroid mice. Recent studies have provided several important clues to address this issue. Another current area that needs attention is the effect of endocrine disruptors on brain development. Since the molecular structures of TH and several endocrine disrupting chemicals are similar, the effect of such chemicals on brain may be exerted at least in part through the TH system. Recent studies have shown the possible interaction between TR and such chemicals. Overall, this review provides current findings regarding molecular mechanisms on TH action in cerebellar development.

Barosensitive and chemosensitive neurons in the rat medulla: A double labeling study with c-Fos/glutamate, GAD, PNMT and calbindin

The purpose of this study was to survey distribution and density of the barosensitive and chemosensitive neurons in the medulla of rats anesthetized with fentanyl/midazolam, using immunohistochemical methods. After stimulation of the arterial baroreceptor or the chemoreceptor, we identified c-Fos-labeled neurons with immunoreactions to antisera of glutamate. PNMT, GAD and calbindin in the nucleus tractus solitarii (NTS) and the ventrolateral medulla (VLM). The double labeled neurons were located in the medical part of the NTS, and in the lateral part of the paragigantocellular reticular nucleus and the ventral division of the ambiguus nucleus. Main findings were as follows: (1) No significant difference was found in distribution and density of glutamatergic, adrenergic and calbindin-containing neurons between the barosensitive and chemosensitivie types; (2) a few GABAergic neurons were distributed almost evenly in the NTS and VLM, and in these neurons the barosensitive type outnumbered the chemosensitive one; (3) glutamatergic and calbindin-containing neurons were dominant in the NTS; adrenergic neurons in the VLM. (4) as for the adrenergic neurons in the NTS, the chemosensitive type significantly outnumbered the barosensitive one. This study showed that distribution and density of the barosensitive neurons, either glutamatergic, adrenergic, or calbindin-containing neurons, overlapped with those of the chemosensitive corresponding neurons, suggesting presence of the neural matrix of the cardiopulmonary interaction. Exceptionally, the number of the barosensitive GABAergic neurons was significantly larger than that of the chemosensitive GABAergic ones.

Molecular cloning of Rhombex-40 a transmembrane protein from the ventral medullary surface of the rat brain by differential display.

Respiration-related neurons, which detect various chemicals in cerebrospinal fluid, are localized to the ventral medullary surface (VMS). We hypothesized that expression of genes involved in respiratory function is upregulated in the VMS. By differential display, we looked for genes differentially expressed in VMS neurons and cerebral cortical neurons. Seventeen clones of interest were isolated, and sequence analysis revealed that one of these clones encoded a putative transmembrane protein, rhombencephalic expression protein-40 kDa (Rhombex-40). The rat Rhombex-40 was composed of 374 amino acid residues, and the predicted secondary structure displays a signal peptide in the N-terminus and single-pass transmembrane domain in the center of the sequence. An analysis of consensus sequences identified several phosphorylation sites in the intracellular domain. Expression of rat Rhombex-40 mRNA is high in the brain, and low in lung, liver and kidney. No homologous protein sequence was found in database searches. Whereas the biological function of this protein is presently unknown, its structural features and high expression in the brain suggest that Rhombex-40 may function as a novel transmembrane molecule in neural cells of the brain.

First Successful Synthesis of Acryloyl-L-Prolylamide Derivatives

Abstract Acryloyl-L-prolylamide (1b) and its two simple derivatives, acryloyl-L-prolyl-N′, N′-dimethylamide (1a) and acryloyl-L-prolyl-N′-methylamide (1c), were synthesized for the first time.

H+-sensitivity of cultured neurons from the dorsomedial and ventrolateral medulla of neonate rats

The H+-sensitivity of neonate rat cultured neurons derived from the dorsomedial medulla (DMM) containing the nucleus tractus solitarii and the ventrolateral medulla (VLM) was determined by H+-sensitive fluorescent probe BCECF-AM and immunohistochemical methods. Against an extracellular pH as low as 7.2-7.3, H+-sensitivity was verified in 2.6% of the DMM neurons (46/ 1800) and 2.1% of the VLM neurons (38/1800). This H+-sensitive neurons of the DMM were immunoreactive to glutamate (52.4%) and glutamic acid decarboxylase (GAD) (28.6%), while those of the VLM were immunoreactive to glutamate (66.7%) and GAD (33.3%). There was no immunoreactivity to tyrosine hydroxylase, phenylethanolamine-N-methyltransferase or choline acetyltransferase in the H+-sensitive neurons are present in the DMM and VLM besides the ventral medullary surface, the site of the central chemoreceptors.

Ulcerogenic cinchophen induces c-Fos expression in CRH-secreting cells in the PVH of rat brain.

This study aimed at obtaining evidence that cinchophen, an ulcerogenic drug, stimulates the corticotropin-releasing hormone (CRH)-secreting cells in the paraventricular nucleus of the hypothalamus (PVH) to induce c-Fos expression. Without colchicine pretreatment, cinchophen was injected i.p. 60 min before the time of maximum CRH level in the hypothalamus, as decided by radioimmunoassay. Eighty percent of the c-Fos/CRH double-labelled cells were concentrated in the parvicellular subnuclei. In the medial and anterior parvicellular subnuclei, the double-labelled neurones of treated rats significantly outnumbered those of controls. The result shows that cinchophen induces excitation of the CRH-secreting cells.