Hyperpolarizing potentials induced by Ca-mediated K-conductance increase in hamster submandibular ganglion cells

The mechanisms of three types of hyperpolarizing electrogenesis in hamster submandibular ganglion cells were analyzed with intracellular microelectrodes. These included (1) spike-induced hyperpolarizing afterpotential (S-HAP), (2) spontaneous transient hyperpolarizing potential (HP), and (3) the hyperpolarizing (H) phase of postsynaptic potential (PSP). Most of these hyperpolarizing potentials were due to conductance increases and reversed polarity at membrane potential (E_m) between ?70 and ?85 mV, which was close to the K-equilibrium potential. The average resting potential of ganglion cells was ?53 mV. Action potential overshoot increased slightly in high [Ca²⁺]₀ and decreased in low [Ca²⁺]₀. In most neurons action potentials were completely suppressed by 10^?7 M tetrodotoxin (TTX). The S-HAP has an initial component due to delayed rectification and a late component. The late component is enhanced by increasing [Ca²⁺]₀, or by applying Ca-ionophore (A23187), TEA, caffeine, or dibutyryl cyclic (DBc-) AMP; it is suppressed by decreasing [Ca²⁺]₀, or by applying Mn²⁺. Perfusion with Cl^?-free saline reduced membrane potential slightly but did not modify the S-HAP. Depolarizing pulses also induced hyperpolarizing afterpotential (D-HAP), similar to the S-HAP. Spontaneous transient HPs occurred in some neurons at irregular intervals. HPs were insensitive to TTX but were suppressed by Mn²⁺. Caffeine induced low frequency rhythmic HPs in many neurons, often alternating with periods of repetitive spiking. The PSP was a monophasic depolarizing (D-) potential in some neurons, but in others the D-phase was followed by a small H-phase. Perfusion with A23187, caffeine or DBc-AMP increased the H-phase of the PSP. Perfusion with K⁺-free saline or treatment with 10^?5M ouabain did not abolish the H-phase of PSPs. These membrane potential-dependent phenomena appear to be induced mainly by Ca-mediated K-conductance increases. This mechanism contributes to the regulation of low-frequency repetitive firing in submandibular ganglion cells.     

The merR regulatory gene in Thiobacillus ferrooxidans is spaced apart from the mer structural genes

Two distinct merR genes, which regulate expression of the mercuric ion resistance gene (mer), of Thiobacillus ferrooxidans strain E-15 have been cloned, sequenced and termed merR1 and merR2. As a result of gene walking around two merR genes, it was found that these two genes were quite close in distance. The nucleotide sequence of the region (5,001 base pairs; PstI-EcoRI fragment) containing the merR genes was determined. Between the two merR genes, there were five potential open reading frames (ORFs). Two of these were identified as merC genes, and the other three as ORFs 1 to 3. ORFs 1 to 3 show significant homology to merA, tnsA from transposon Tn7, and merA, respectively. Both merR genes consist of a 408 bp ORF coding for 135 amino acids. Their gene products, MerR1 and MerR2, differed at three amino acid positions, and shared 56-57% and 32-38% identity with the MerRs from other Gram-negative and Gram-positive bacteria, respectively. Competitive primer extension analysis revealed that both regulatory genes were expressed in the host cells. These merR genes were located more than 6 kb from either end of the mer structural genes (merC-merA). This is the first example of merR being separated from the mer structural genes. The two merC genes, each of which coded for a 140-amino-acid protein, appeared to be functionally active because Escherichia coli cells carrying these merC genes on plasmid vectors showed hypersensitivity to HgCl2. However, ORFs 1 and 3, which were homologous to merA, seemed to be inactive both structurally and enzymatically. The gene arrangement in this region took on a mirror image, with the truncated tnsA as the symmetrical centre. It is suggested that the Tn7-like factor may have participated in gene duplication events of the mer region, and in its chromosomal integration.     

Growth and survival rate of the larvae ofHynobius nebulosus Tokyoensis Tago (Amphibia,Hynobiidae)

Growth and population density of the larvae, Hynobius nebulosus tokyoensisTago , were estimated in a small pond within the study site settled in Habu village of Hinodemachi, a suburb of Tokyo City, during the period from 1975 to 1980. The mortality factors which influenced the survival rate of larvae were also evaluated from the ecological point of view. Laboratory experiments on the growth of larvae and predation by newts were conducted in pararell with the field survey. The results showed that growth rate of larvae under the natural condition was very slow, as compared with that under the laboratory condition with sufficient food supply, and mean body size at metamorphosis was negatively correlated with the density at that time. This suggested that food resources were in short supply in the pond, and there occurred a severe intraspecific competition for food among larvae. The mortality rate of larvae was so high, 80–99% in each year, and the density of larvae survived until metamorphosis varied so greatly from year to year that the larval stage was the most important stage throughout the life cycle to the maintenance of a population for this salamander. The most important factors which contributed to this high mortality were the predation by the newt, Triturus pyrrhogaster pyrrhogasterBoie , and cannibalism. From the laboratory experiment, it was found that predators could attack only small larvae successfully, and successful attack rate decreased sharply as larvae grew larger. This relationship resulted in the characteristic L-shaped pattern of survivorship curve of larvae; that is, heavy mortality just after hatching period.     

Decidual cells are the initial target of polyriboinosinic–polyribocytidylic acid in a mouse model of maternal viral infection

BackgroundMaternal immune activation has been implicated in the pathophysiology of neurodevelopmental disorders such as autism spectrum disorders caused by maternal infection. It has been suggested that the placental origin of inflammatory cytokines leads to neurodevelopmental disorders. However, the identity of the initial immune-activated site in the placenta, in response to maternal viral infection, is not clear.MethodsBy cross-breeding male enhanced green fluorescent protein (EGFP) transgenic mice with wild-type females, the placental tissues of maternal origin can be distinguished from those of paternal origin by EGFP expression. Using this method, at embryonic day (E) 12.5, dams were administered an intraperitoneal polyriboinosinic–polyribocytidylic acid (poly [I:C]) injection. We quantitatively analyzed the levels of phosphorylated interferon (IFN) regulatory factor 3 (pIRF3) in the placenta, and investigated the distribution of pIRF3 positive cells.ResultsWe show that maternally derived decidual cells are the initial target of maternal poly (I:C) through the toll-like receptor 3/TIR-domain-containing the adapter-inducing interferon-β signaling pathway. We also show that the expression of interferon-β was upregulated in the placenta after maternal injection with poly (I:C).ConclusionThese results suggest that maternally derived decidual cells are the initial target of maternal poly (I:C) and that this innate immune response is likely associated with a state of maternal immune activation.