Cell type-specific expression of the human renin gene.

We have previously produced transgenic mice carrying the human renin gene, whose expression is regulated in a tissue-specific manner. In the present study, we further characterized expression of the transgene. Northern blot analysis showed that the human renin gene is expressed in the kidney but not in the liver of two lines of transgenic mice with 10 and 50 copies of the transgene, suggesting that the integrated copy number of the human renin gene does not influence the dominant-renal expression pattern. Immunohistochemical study using a monoclonal antibody specific for human renin demonstrated that expression of human renin in the transgenic mouse kidney is confined to the epithelioid juxtaglomerular cells. Transfection experiments indicated that the chloramphenicol acetyltransferase fusion gene containing the 3-kb upstream sequences of the renin gene is activated only in human epithelioid embryonic 293 cells derived from kidney but not in human HepG2 cells from liver. These findings suggest that transfer of the cloned renin gene into mice and in vitro cultured cell lines can give rise to cell type-specific expression.     

Mycobacteria in the light of modern genetics development

It is generally assumed that genetic research of mycobacteria is delayed as compared with other, more commonly used, bacterial models, particularly in the field of genetic transfers. In the field of mutagenesis the problems have been studied to such an extent that replication maps of the chromosome of M. phlei and M. tuberculosis H37 Rv have already been constructed and a new model of the cell cycle of bacteria exhibiting a slow growth rate has been worked out. When the problems of mycobacterial genetics are looked upon in the light of gene manipulations it may be concluded that mycobacteria belong to a few models whose genes are used for cloning and that problems of practical significance will be studied by means of the most modern approaches.     

Pigmentation changes in Brevibecteria induced by mutagenic treatment

Inducible pigmentation changes were observed in pigmented strains of Brevibacterium sp. M27 and B. flavum treated with N-methyl-N'-nitro-N-nitrosoguanidine. The highest frequency of induction was reached already at a survival of 30-40% with the maximal yield of 6-10%. As compared with the initial yellow colour, three new pigmentation types, viz. white, pink and orange, were observed. The yellow pigmented parent strains are most resistant to the lethal effects of UV radiation. By selecting pigmented mutants of all types on media containing antibiotics it was possible to obtain strains that were resistant either to tetracycline or to streptomycin. Auxotrophic pigmented mutants were also isolated. In multiple mutant strains of Brevibacterium sp. M27 a number of strainsexhibited a changed L-lysine production. In some strains the production was variable, whereasother strains did not produce L-lysine at all and stains with a limited production of other amino acids were also detected.     

Protoplast forming ability in the genusBrevibacterium

Formation of protoplasts and their reversion were followed in 7 strains of brevibacteria. The formation of protoplasts and their reversion differed both between various species of brevibacteria and between various mutant strains of the same species.     

Mobilization of the transposable element mdg4 by hybrid dysgenesis generates a family of unstable cut mutations in Drosophila melanogaster

Summary A family of unstable mutations at the cut locus in Drosophila melanogaster was obtained under the conditions of hybrid dysgenesis (Gerasimova 1981, 1982). The in situ hybridization experiments have shown that, in the original unstable ct ^MR2 mutation, the 7B region of the X chromosome (where cut is located) contains a mobile dispersed genetic element, mdg4. All other unstable ct mutations derived from ct ^MR2 including visible and lethal alleles and unstable ct ⁺ reversions, also contain mdg4 in the 7B region. The X chromosomes of the parent strain (wild type) do not contain mdg4 at all. All stable revertants derived from ct ^MR2, from other unstable ct mutations, or from ct lethals lost mdg4 from the 7B region. The ct ^MR2 X chromosome does not contain P-elements, although a few copies are present in the autosomes. The instability of the ct ^MR2./ct ^MR2 strain remained at a high level for 50 generations (1.5 years) and then rapidly decreased. A new cross with an MRh12/Cy strain (originally used for dysgenesis induction and containing a number of P-elements) increased the instability to a level exceeding the original one. The data strongly suggest that unstable ct mutations in our system are induced by transpositions of mdg4, possibly activated by P-elements.     

The mating system ofTokunagayusurika akamusi (Diptera; Chironomidae): II. Experimental analysis of male mating behaviour at the resting place

Mating behaviour ofTokunagayusurika akamusi searching at the resting place (lakeside vegetation, although mating also occurs in the air by swarming) was investigated at the shore of Lake Biwa. Coinciding with the arrival of newly emerged adults, previously emerged males walked about to search for mates at the resting place. When a searching male came into contact with a fresh (newly emerged) individual, he attempted to copulate with it regardless of its sex. Consequently, males copulated with fresh females, which were more likely to be virgin at the resting place, suggesting that the first male might contribute best to the fertilization.     

Mating system ofTokunagayusurika akamusi (Diptera: Chironomidae): I. Copulation in the air by swarming and on the ground by searching

Observations on the mating system of the midge,Tokunagayusurika akamusi, revealed mating to occur both in the air by swarming and on the ground by searching. At the shores of Lake Biwa, midges appeared from November to early December. Newly emerged adults arrived at the resting place, lakeside vegetation, in the morning, during which time a number of males also walked about in search of mates. Many copulating pairs were observed at the resting place. Huge swarms occurred chiefly before sunset but the frequency of copulation observed in the swarm was extremely low. It is likely that, in the Lake Biwa population, the proportion of females inseminated by searching males at the resting place was much larger than that by swarming males in the air. Furthermore, by searching, males copulated with younger females than by swarming. The differences between the searching and swarming tactics are discussed.     

Formation of partially phosphorylated phosphorylase in isoproterenol stimulated rat hearts

Summary Phosphorylase ab hybrid was demonstrated in perfused rat hearts and during the in vitro conversion of purified rat heart phosphorylase b. Phosphorylase ab hybrid was determined in rat heart extracts by the activating effect of AMP in the presence of caffeine. These results were confirmed by the quantitative determination of incorporated ³²P in vitro and through the characteristic inhibition of ab hybrid by glucose-6-phosphate.As shown by our results, in aerobically perfused control hearts only the ab hybrid represents the active form of phosphorylase, its activity reaching about 20% of the total. In response to isoproterenol (5–1000 ng), the amount of ab hybrid rose to about 30–40%, preceding the rise of the a form, which increased in a dose-dependent manner up to 45% of the total.The great sensitivity of the ab form to AMP activation and glucose-6-phosphate inhibition supports its physiological significance in heart under in vivo conditions as well. Our results strongly suggest that the activity ratio -AMP/+AMP reflects rather the percentage ratio of phosphorylated subunits than that of the activated (partially or totally phosphorylated) phosphorylase molecules.     

Glomerular mesangium as an effector locus for the tubuloglomerular feedback system and renal sympathetic innervation

To define the effector loci for the tubuloglomerular feedback system, the determinants of the single-nephron glomerular filtration rate (SNGFR) were assessed in Munich-Wistar rats by direct glomerular puncture during perfusion of Henle's loop with isotonic Ringer's solution at rates of 0 and 40 nl/min. At the higher flow rate, SNGFR averaged only approximately 65% that measured during the lower flow rate. Whereas mean glomerular capillary hydraulic pressure was unaffected, both glomerular plasma flow rate and ultrafiltration coefficient Kf were found to decrease significantly in response to increase in loop perfusion rate, thereby accounting for the measured reduction in SNGFR. These changes were associated with increased afferent (RA) and efferent (RE) arteriolar resistances. Based on the close anatomic contact between mesangial cells and these arterioles, a single effector mechanism channeled through mesangial contractility is suggested to account for the observed reduction in Kf and increase in RA and RE. Mesangial contractility appears to be under sympathetic nerve control. In our recent micropuncture study with Munich-Wistar rats, a marked reduction in SNGFR was observed during high-frequency stimulation (5 Hz) of the renal nerve. This reduction in SNGFR was accompanied by a marked fall in Kf and increase in RA and RE. When kidneys were perfusion-fixed during high-frequency stimulation, a marked reduction in the number of open channels was demonstrated together with marked narrowing of afferent and efferent arterioles. These observations are consistent with the view that sympathetic innervation of mesangium may modulate GFR through its ability to regulate mesangial contractility.