The effect of cycloheximide on the entry into the S period of the nuclei in heterodikaryons]

Serum-deprived (0.2%) resting NIH 3T3 mouse fibroblasts were fused with serum-stimulated (10%) proliferating cells to elucidate mechanisms of entering into S-period operating in the nuclei of the heterokaryons under the effect of cycloheximide--an inhibitor of protein synthesis. Using radioautography DNA synthesis was investigated in mono-, homo- and heterodikaryons. After short (0.5-3.0 h) depressing of protein synthesis, the nuclei of stimulated cells in heterokaryons were found to enter into S-period. Under these conditions no induction of DNA synthesis was found in the nuclei of resting cells in heterodikaryons. In other experiments, resting cells were under the effect of cycloheximide during 2-4 h before the fusion, that led to a great induction of DNA synthesis in the nuclei of these cells in heterodikaryons. The data obtained are consistent with the idea of fibroblast transition to the rest under the action of labile proteins-repressors.     

Electrophysiological analysis of corticofugal influences on preoptic neurons

Evoked potential (EPs) and responses of the medial (MPO) and lateral (LPO) preoptic region (RPO) and adjacent structures of the hypothalamus to stimulation of the prefrontal (area 8) and cingulate (area 24) cortex, piriform lobe (periamygdaloid cortex), and hippocampus (area CA3) were investigated in acute experiments on cats under ketamine anesthesia. The most pronounced EPs were observed in the RPO after stimulating the piriform and cingulate cortex. A close relation was found between neuronal responses and EP components. The majority of neurons responding to stimulation of various cortical structures were localized in the LPO, where primarily excitatory responses dominate. The MPO contained somewhat fewer neurons responding to cortical stimulation, and the dominant response here was primarily inhibitory. The ratio of inhibitory and excitatory responses in the LPO was 0.6:1 and in the MPO 5.8:1. Primarily in-inhibitory responses dominated also in the LPO zone adjacent to the bed nucleus stria terminalis (BST) and primarily excitatory in the region surrounding the supraoptic nucleus (SO) (respective ratios 4.9:1 and 0.7:1). The RPO is a broad convergence zone, where 3/4 of the neurons responded to stimuli of two and more cortical regions.A. M. Gorky Medical Institute, Ukrainian Minstry of Health, Donetsk. Translated from Neirofiziologiya, Vol. 23, No. 6, pp. 709–719, November–December, 1991.     

rRNA-protein neighbourhood in Escherichia coli 70 S ribosomes and 70 S initiation complex. Probing by bifunctional Pt(II)-containing reagent

The cleavable homobifunctional reagent dichloro[N,N,N',N'-tetrakis(2-aminoethyl)-1,6-hexamethylenediamminedi platinum (II)] dichloride was used for studying rRNA-protein cross-links in free 35S-labelled 70 S ribosomes and within initiation complex ribosome.AUGU6.fMet-tRNA(fMet). It was shown that the sets of proteins cross-linked to 16 S and 23 S rRNA in free 70 S ribosomes and in 70 S initiation complex do not differ significantly. The authors are the first to demonstrate most of the 23 S rRNA-protein cross-links and some 16 S rRNA-protein cross-links, in particular those with L7/L12 protein.     

The channel-forming component of the Theridiidae spider venom neurotoxins

It is known that Steatoda (Lityphantes) paykulliana and Latrodectus mactans tredecimguttatus spider venoms are toxic to mammals and insects. These venoms act presynaptically eliciting massive release of transmitters. They also form channels in bilayer lipid membranes (BLM) that are selective for cations. Venoms of both spider species were fractionated by gel filtration on a Sephadex G-100 column. The fraction obtained were tested on neuromuscular preparations of frog and locust and on BLM. A fraction of low molecular weight components (about 5000 daltons and less) was disclosed. This fraction showed presynaptic and channel-forming effects similar to those of crude venoms and of high molecular weight toxin fractions, obtained simultaneously from these venoms. It was shown that channels formed in BLM by crude venoms and its different fractions are identical. Also, it was found that the low molecular weight channel-forming component is a construction element of high molecular weight toxins. On the basis of data obtained a toxin structure model of the Theridiidae family spider venoms was proposed.     

Properties of the channels induced by Lityphantes paykullianus venom

It was found that common venom of Lityphantes paykullianus is capable of forming conductivity channels on bilayer phospholipid membrane. The values of channel conductivity are remarkably distinguished in the solutions of different alkaline metals. The selectivity of channels is mainly cationic at pH 7.5 with weak discrimination of ions few tell one from the other by size. These results indicate that there is an active component in the common venom accountable for channels formation.     

Electrophysiological analysis of corticofugal connections with the medial dorsal thalamic nucleus

In acute experiments on cats anesthetized with pentobarbital and chloralose, focal responses were recorded to study projections of various parts of the orbitofrontal cortex and cortex of the temporal pole in the region of the medial dorsal nucleus of the thalamus and interaction in this nucleus between stimuli arriving from the medio-basal portions of the neocortex. Different parts of the orbitofrontal cortex were found to have local projections in the medial dorsal nucleus so arranged that the rostral zones of the cortex send stimuli to the medio-dorsal portions of the nucleus, whereas regions of the cortex radiating fanwise from the pole in dorsal and caudal directions are arranged in the lateral and basal portions of the nucleus. The cortex of the temporal pole has relatively diffuse projections in the medial part of the medial dorsal nucleus. Stimuli reaching the medial dorsal nucleus from the basal structures of the neocortex (temporal pole) were shown to facilitate response to stimulation of the orbitofrontal cortex. Meanwhile, stimulation of this region of the cortex depresses the receptive capacity of the nucleus for impulses arriving from the temporal cortex.     

Organization of neurons forming descending corticohypothalamic pathways

Retrograde horseradish peroxidase axonal transport techniques were used for quantifying the spatial organization of cortical neurons constituting descending fiber systems running to hypothalamic structures. This revealed the diversity of corticohypothalamic connections: the prefrontal cortex exerts a more marked effect on the hypothalamus than other cortical regions of the limbic system. Corticohypothalamic cells form neuronal groups at different frontal levels of the prefrontal cortex and cingulate gyrus; shape and orientation of cell groups are determined by the topography of predominant axonal systems travelling to the hypothalamus. Various forms of cell combinations occur within neuronal populations. Neuronal arrays may underlie the organizational principle of direct corticohypothalamic pathways.A. M. Gor'kii Medical Institute, Ministry of Public Health of the Ukrainian SSR, Donetsk. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 435–441, June–July, 1990.     

Anonymous nuclear DNA markers in the American oyster and their implications for the heterozygote deficiency phenomenon in marine bivalves

A puzzling population-genetic phenomenon widely reported in allozyme surveys of marine bivalves is the occurrence of heterozygote deficits relative to Hardy-Weinberg expectations. Possible explanations for this pattern are categorized with respect to whether the effects should be confined to protein-level assays or are genomically pervasive and expected to be registered in both protein- and DNA-level assays. Anonymous nuclear DNA markers from the American oyster were employed to reexamine the phenomenon. In assays based on the polymerase chain reaction (PCR), two DNA-level processes were encountered that can lead to artifactual genotypic scorings: (a) differential amplification of alleles at a target locus and (b) amplification from multiple paralogous loci. We describe symptoms of these complications and prescribe methods that should generally help to ameliorate them. When artifactual scorings at two anonymous DNA loci in the American oyster were corrected, Hardy-Weinberg deviations registered in preliminary population assays decreased to nonsignificant values. Implications of these findings for the heterozygote-deficit phenomenon in marine bivalves, and for the general development and use of PCR-based assays, are discussed.