Catecholaminergic innervation of GRF-containing neurons in the rat hypothalamus revealed by electron-microscopic cytochemistry

Summary The Catecholaminergic innervation of neurons containing growth hormone-releasing factor (GRF) was examined by use of a method which combined either 5-hydroxydopamine (5-OHDA) uptake or autoradiography after intraventricular injection of ³H-noradrenaline with immunocytochemistry for GRF in the same tissue sections at the electron-microscopic level. In the ventrolateral part of the arcuate nucleus of the rat hypothalamus a large number of immunonegative axon terminals were found to make synaptic contact with GRF-like immunoreactive (GRF-LI) cell bodies and processes. ³H-noradrenaline autoradiography or 5-OHDA-labeling combined with GRF immunocytochemistry revealed that axon terminals labeled with ³H-noradrenaline or 5-OHDA make synaptic contact with the GRF-LI nerve cell bodies and processes. These findings indicate that catecholamine-containing neurons innervate GRF neurons to regulate GRF secretion via synapses in the rat arcuate nucleus.     

IS421, a new insertion sequence in Escherichia coli

The nucleotide sequence of a new insertion sequence (IS) in Escherichia coli, IS421, was determined. It is 1340 bp long and contains inverted repeats of 22 bp at its termini. It is flanked by 13 bp direct repeats apparently generated upon insertion. There are two ORFs longer than 200 bp in IS421. One can encode a polypeptide of 371 amino acids (aa) and the other, which is on the other strand, can encode a polypeptide of 102 aa. The C-terminal part of the 371 aa polypeptide shows some homology to that of transposases encoded in some other known IS elements. The copy number of IS421 in chromosomal DNA was 4 for E. coli K-12 and B, and 5 for E. coli C, as determined by the Southern hybridization of restriction fragments.     

Cell swelling induced by the permeant molecules urea or glycerol induces immediate high amplitude thyrotropin and prolactin secretion by perifused adenohypophyseal cells

The permeant molecules, urea and glycerol, evoked a prompt secretory burst of TSH and PRL when added to the extracellular medium of acutely dispersed anterior pituitary cells. Secretion of both hormones was proportional to the concentration of urea or glycerol between 26 and 104 mM (r greater than 0.89, P less than 0.001). Equivalent concentrations of the impermeant molecule, mannitol, did not induce secretion. The acute TSH and PRL secretory responses to TRH, hyposmolarity, and permeant molecules were qualitatively indistinguishable. These data support our hypothesis that cell swelling and resultant plasmalemma expansion is a potent inducer of hormone secretion. Since the secretory response to permeant molecules was not reduced in a Ca2+-free medium containing 0.1 mM EGTA, an increase in Ca2+ transport across the plasmalemma to raise cytosol Ca2+ concentration does not appear involved.     

Effect of cell cholesterol content on apolipoprotein B secretion and LDL receptor activity in the human hepatoma cell line, HepG2

Human hepatoma HepG2 cells were used to study the effects of cholesterol loading and depletion on apolipoprotein B (apoB) secretion and low-density lipoprotein (LDL) receptor activity. Exposure of HepG2 cells to cholesterol and oleic acid, which elevated intracellular cholesterol levels, stimulated apoB secretion and reduced receptor-mediated uptake of LDL, whereas recombinant complexes of apolipoprotein A-I with dimyristoylphosphatidylcholine, which depleted the cellular cholesterol pool, inhibited apoB secretion and up-regulated LDL receptors. Significant negative correlation (r = -0.92, P less than 0.001) between the levels of apoB secretion and LDL uptake was found. These data suggest that the cholesterol content of the cells may induce concomitant changes in apoB secretion and LDL receptor activity.     

A real time learning algorithm for recurrent analog neural networks

A new learning algorithm is described for a general class of recurrent analog neural networks which ultimately settle down to a steady state. Recently, Pineda (Pineda 1987; Almeida 1987; Ikeda et al. 1988) has introduced a learning rule for the recurrent net in which the connection weights are adjusted so that the distance between the stable outputs of the current system and the desired outputs will be maximally decreased. In this method, many cycles are needed in order to get a target system. In each cycle, the recurrent net is run until it reaches a stable state. After that, the weight change is calculated by using a linearized recurrent net which receives the current error of the system as a bias input. In the new algorithm the weights are changed so that the total error of neuron outputs over the entire trajectory is minimized. The weights are adjusted in real time when the network is running. In this method, the trajectory to the target system can be controlled, whereas Pineda's algorithm only controls the position of the fixed point. The relation to the back propagation method (Hinton et al. 1986) is also discussed.     

Holocene sedimentary history of some coastal plains in Hokkaido,Japan IV. Diatom assemblages in the sediments from Kushiro moor (2)

Diatom assemblages of sediments obtained from three sites on Kushiro Moor were analyzed to investigate the Holocene sedimentary history. The results showed that: 1) The Takkobu site was originally at the bottom of the paleo-Kushiro Bay, and after-wards the paleo-Takkobu Lagoon developed, became sealed off, and changed to a freshwater lake. The succession to peat moor probably began about 2000 yr B.P. at the Takkobu site. 2) The Tsurui site was originally at the bottom of the paleo-Kushiro Bay, then changed to the paleo-Kushiro Lagoon and became peat moor as a result of the first Holocene regression, which finished about 3600 yr B.P. The site then returned to a brackish lake again, probably due to the second Holocene transgression between 3600 and 3000 yr B.P., thereafter passing through brackish lake and freshwater lake stages, and eventually becaming peat moor at about 2000 yr B.P., 3) At the Chuo site, the second paleo-Kushiro Bay developed again as a result of the second Holocene transgression, which finished about 3000 yr B.P. Thereafter, brackish or freshwater lakes, rivers, and then peat moor developed in the central area of Kushiro Moor. 4) The second marine diatom zone (MD₂ Zone), which indicates the second Holocene transgression, complete by about 3000 yr B.P., is detected only at the Chuo site in the central area of Kushiro Moor.     

A quantitative comparison of foliage development among allopatric ferns,Dryopteris crassirhizoma,D. coreano-montana andD. filix-mas

A quantitative comparison was conducted on the foliage development during sporophyte development of three allopatric ferns in cool temperate and subalpine regions of Hokkaido and Tirol, European Alps. The foliage development ofDryopteris crassirhizoma, D. coreano-montana andD. filix-mas was quantitatively described by the leaf development (NV, number of veins); NV correlates the leaf-shape complexity from a circle (DI, L/2(3.14×S)^1/2). Nearly similar patterns were detected on frequency distribution of fertile leaves, fertility increase and number of leaves in threeDryopteris ferns which exhibit funnel-shaped foliage arrangements in mature sporophyte. No difference was found in number of leaves, maximum NV, fertility rate and leaf-shape parameters among three ferns. A positive difference was found only on changes in order of pinnae with maximum number of costa branches (NVP) and the DI of outline of pinnae betweenD. crassirhizoma andD. filix-mas. These allopatricDryopteris ferns seem to have a similar foliage structure, in spite of some sympatricDryopteris ferns capable of producing putative hybrids (D. austriaca andD. amurensis; D. tokyoensis andD. monticola) having different foliage structures in Hokkaido. Contribution No. 3346 from the Institute of Low Temperature Science, Hokkaido University.     

Processing, secretion, and biological properties of a novel growth factor of the fibroblast growth factor family with oncogenic potential.

We recently reported that the protein encoded in a novel human oncogene isolated from Kaposi sarcoma DNA was a growth factor with significant homology to basic and acidic fibroblast growth factors (FGFs). To study the properties of this growth factor (referred to as K-FGF) and the mechanism by which the K-fgf oncogene transforms cells, we have studied the production and processing of K-FGF in COS-1 cells transfected with a plasmid encoding the K-fgf cDNA. The results show that, unlike basic and acidic FGFs, the K-FGF protein is cleaved after a signal peptide, glycosylated, and efficiently secreted as a mature protein of 176 or 175 amino acids. Inhibition of glycosylation impaired secretion, and the stability of the secreted K-FGF was greatly enhanced by the presence of heparin in the cultured medium. We have used the conditioned medium from transfected COS-1 cells to test K-FGF biological activity. Similar to basic FGF, the K-FGF protein was mitogenic for fibroblasts and endothelial cells and induced the growth of NIH 3T3 mouse cells in serum-free medium. Accordingly, K-fgf-transformed NIH 3T3 cells grew in serum-free medium, consistent with an autocrine mechanism of growth. We have also expressed the protein encoded in the K-fgf protooncogene in COS-1 cells, and it was indistinguishable in its molecular weight, glycosylation, secretion, and biological activity from K-FGF. Taken together, these results suggest that the mechanism of activation of this oncogene is due to overexpression rather than to mutations in the coding sequences.