RIBONUCLEIC ACID SYNTHESIS DURING MITOSIS AND MEIOSIS IN THE MOUSE TESTIS

The pattern of ribonucleic acid synthesis during germ cell development, from the stem cell to the mature spermatid, was studied in the mouse testis, by using uridine-H³ or cytidine-H³ labeling and autoradiography. Incorporation of tritiated precursors into the RNA occurs in spermatogonia, resting primary spermatocytes (RPS), throughout the second half of pachytene stage up to early diplotene, and in the Sertoli cells. Cells in leptotene, zygotene, and in the first half of pachytene stage do not synthesize RNA. No RNA synthesis was detected in meiotic stages later than diplotene, with the exception of a very low rate of incorporation in a fraction of secondary spermatocytes and very early spermatids. At long intervals after administration of the tracer, as labeled cells develop to more mature stages, late stages of spermatogenesis also become labeled. The last structures to become labeled are the residual bodies of Regaud. Thus, the RNA synthesized during the active meiotic stages is partially retained within the cell during further development. The rate of RNA synthesis declines gradually with the maturation from type A to intermediate to type B spermatogonia and to resting primary spermatocytes. "Dormant" type A spermatogonia synthesize little or no RNA. The incorporation of RNA precursors occurs exclusively within the nucleus: at later postinjection intervals the cytoplasm also becomes labeled. In spermatogonia all mitotic stages, except metaphase and anaphase, were shown to incorporate uridine-H³. RNA synthesis is then a continuous process throughout the cell division cycle in spermatogonia (generation time about 30 hours), and stops only for a very short interval (1 hour) during metaphase and anaphase.     

CHLORAMPHENICOL IN THE TREATMENT OF TYPHOID FEVER

Six patients with typhoid fever were treated with chloramphenicol. The excellent clinical response in four cases suggests that chloramphenicol is the drug of choice in the treatment of this disease. In one case in which clinical relapse occurred, there was good response to re-treatment. One patient, critically ill, in a typhoid state, and treated late in the course of the disease, died without beneficial effect from chloramphenicol, but the patient had been unable to retain the drug because of vomiting.     

人胚肝发育过程中染色质组份的变化

从14-,17-,21-,27-,34-,及38-周令的人胚肝细胞核分离出柒色质,分别对其中的RNA、DNA、组蛋白(HP)及非组蛋白(NHP)进行测定。在胚胎发育过程中肝柒色质HP/DNA比值变化不大。但是,NHP/DNA与NHP/HP比值发生显著改变。人胚肝NHP量的变化一直保持在整个胚胎发育过程中,NHP量的高峰位于21-及34-周。用SDS-聚丙烯酰胺凝胶板电泳分析处于不同发育阶段的人胚肝总染色质蛋白。电泳图谱显示出染色质NHP组分在质与量上有所改变。     

MICRONUCLEAR RIBONUCLEIC ACID IN TETRAHYMENA PYRIFORMIS

The presence of RNA in the micronucleus of Tetrahymena pyriformis was detected by electron microscope radioautography after incubation with tritiated precursors. The specificity of RNA labeling was shown by ribonuclease digestion. The period of appearance of labeled RNA in the micronucleus is approximately coincident with the DNA synthesis period for the micronucleus. Pulse-chase experiments showed that the micronuclear RNA disappears during the interphase period. The experiments do not distinguish whether the micronuclear RNA is synthesized in situ or acquired by migration from the macronucleus. In either case it is notable that the appearance of labeled RNA is detected in the micronucleus only during the micronuclear S phase.     

CHANGE IN ENZYME ACTIVITY AND RIBONUCLEIC ACID CONCENTRATION WITHIN THE EPIDERMAL CELL OF THE RAT DURING THE GROWTH STAGE OF THE HAIR CYCLE

The rat hair cycle produces rapid anatomical and physiological changes in the entire skin. By the 4th day of the cycle, the epidermis has doubled in thickness. The present study shows that, during this proliferative period, there is a correlation between the biological state (growth) and the molecular composition (enzyme concentrations) of the epidermal cell. In proliferated 4th day epidermis there has occurred not only an increase in the number of viable cells but also an increase in succinic dehydrogenase and cytochrome oxidase activity and ribonucleic acid (RNA) content in the viable epidermal cell. Moreover, RNA content and enzyme activities appear to be correlated. The significance of these findings is discussed in the light of recent evidence supporting the relationship between RNA and enzyme synthesis and the role of hormones in the induction of enzyme adaptation.     

THE INTRACELLULAR DISTRIBUTION AND HETEROGENEITY OF RIBONUCLEIC ACID IN STARFISH OOCYTES

A study has been made of the content and composition of RNA in cytoplasm, nucleoplasm, and nucleoli from growing oocytes of the starfish Asterias rubens. The determinations were carried out, using ultramicrochemical methods, on units isolated by microdissection from fixed sections. Macrochemical and interferometric control experiments show that RNA can be quantitatively evaluated in this way. The results show that the growing oocyte represents a system in which the relations between the quantities of nucleolar, nucleoplasmic, and cytoplasmic RNA undergo great changes. These changes are continuous for nucleolar and cytoplasmic RNA so that their amounts may be predicted from the size of the cell. Nucleoplasmic RNA, on the other hand, shows great variations among different cells, independent of cell size. Purine-pyrimidine analyses show that each cell component contains an RNA which differs significantly from that of the other two. Cytoplasmic and nucleolar RNA are closely related, the only difference being a slightly higher guanine/uracil quotient for the nucleolar RNA. They are both of the usual tissue RNA type, i.e., they show a preponderance of guanine and cytosine over adenine and uracil. Nucleoplasmic RNA deviates grossly from the RNA of the other two components. Here the concentrations of adenine and uracil are higher than those of guanine and cytosine, respectively. This RNA consequently shows some resemblance to the general type of animal DNA although the purine/pyrimidine ratio is far from unity. Our data favor a nucleolar origin for the stable part of the ribosomal RNA and a nucleoplasmic one for the unstable part (the messenger RNA).     

THE CONTENT AND RELATIVE BASE RATIOS OF RIBONUCLEIC ACID IN AMOEBA

The amount and relative base ratios of ribonucleic acid (RNA) in the nucleus and cytoplasm of Amoeba proteus and A. dubia, and of homospecies cells obtained by nuclear transfer with A. proteus, have been determined by microelectrophoresis. In A. proteus the average amounts of RNA in the nucleus and the cytoplasm were 134. micromicrograms and 2520. micromicrograms; in A. dubia the averages for the nucleus and cytoplasm were 67. micromicrograms and 1427. micromicrograms. The relative base ratio of RNA of the nucleus is similar to that of the RNA of the cytoplasm within a species, but the two species differed in this respect. Homospecies nuclear transfer did not affect the relative base ratio or amount of RNA.     

CHANGES IN THE BASE COMPOSITION OF NUCLEAR RIBONUCLEIC ACID OF NEURONS DURING A SHORT PERIOD OF ENHANCED PROTEIN PRODUCTION

Nuclei from isolated nerve cells were sampled by microdissection. The content and composition of the nuclear RNA was studied and compared with that of the cytoplasmic RNA of Deiters' nerve cells of rabbits. Analyses were made of control nerve cells and of cells in which an enhanced RNA and protein production had been induced by chemical means, tricyano-amino-propene, for 60 minutes. The nuclear RNA content of the control nerve cells was 56 µµg, i.e. 3 per cent of the total RNA content of the nerve cell. The base ratios were: adenine 21.3, guanine 26.6, cytosine 30.8, uracil 21.3. Purine-pyrimidine analyses showed that the nuclear RNA differed significantly from the cytoplasmic RNA in having higher adenine and uracil values. The guanine and cytosine values were high, however, and the ratio G/C was 0.86 as compared with 1.16 for the cytoplasmic RNA. The composition of the nuclear RNA was interpreted as reflecting the extraordinarily strong development of the nucleolus in these neurons. During the 60 minutes of enhanced neuronal RNA production (+25 per cent) the guanine value increased and the uracil value decreased significantly in the nuclear RNA. In the cytoplasmic RNA the guanine value also increased although not so much as the nuclear guanine. The cytoplasmic cytosine value decreased. The result indicated that the production of the characteristic cytoplasmic RNA had been influenced by the change in the nuclear RNA     

THE NATURE AND PROCESSING OF RIBOSOMAL RIBONUCLEIC ACID IN A DINOFLAGELLATE

Certain features of the dinoflagellate nucleus suggest that it represents a primitive form of eukaryotic nucleus. For this reason, it was of interest to characterize dinoflagellate ribosomal RNA (rRNA) and its mode of synthesis to determine if it also deviated from typical eukaryotic patterns. Gyrodinium cohnii was chosen for this examination. Gyrodinium ribosomal RNA species are 16 and 25s as judged by their sedimentation velocities in isokinetic sucrose gradients. These values are typical of higher plants. In addition, the RNA cosedimented precisely with rRNA from the ciliate Tetrahymena. Nucleotide ratio analyses revealed a GMP + CMP content of 46% for both species of rRNA. The kinetics of incorporation of a radioactive precursor into ribosomal RNA have also been studied, and it seems likely that the maturation of rRNA starts with the synthesis of a 38s molecule. This serves as precursor to the 16s species, and, after a 27s intermediate, the 25s ribosomal component. The process is similar to that in other eukaryotes. The structure of the nucleolus has also been examined, and is seen to be typically eukaryotic.     

THE RELATION BETWEEN THE INTRACELLULAR RIBONUCLEIC ACID DISTRIBUTION AND AMINO ACID INCORPORATION IN THE LIVER OF THE DEVELOPING CHICK EMBRYO

The RNA-P and DNA-P content of the nucleus and the RNA-P content of the whole cell of the livers of 8- to 20-day chick embryos and of adult fowls have been determined. The DNA-P content of the liver nuclei was slightly higher in the 8- and 10-day embryo than in all the other stages examined. A significant decrease in the RNA content of the cell occurred during embryonic development. The RNA content of the adult cell was the same as that of the 14- to 16-day embryo. The proportion of the cellular RNA contributed by the nucleus also decreased during development. In respect to both nuclear RNA content and distribution of RNA between nucleus and cytoplasm, the adult resembled the 8- to 12-day embryo. Examination of the fine structure of the cell showed that, as development progressed, free ribosomes decreased in number and the rough membranes increased. Slices of 8-, 14-, and 20-day embryonic livers and of adult livers were incubated with ¹⁴C-leucine, and the amount of labeled amino acid incorporated into whole tissue protein and into the proteins of the subcellular fractions was measured. Embryonic liver incorporated ¹⁴C-leucine 15 to 30 times more rapidly than adult liver. The microsomal protein was always more highly labelled than the protein in any other subcellular fraction; however, in the 8-day embryonic and the adult liver the proportion of total counts found in the nuclear fraction was considerably higher than in the 14- or 20-day embryonic liver. The significance of an apparent correlation between the proportion of the cell's RNA contributed by the nucleus and the proportion of total counts in the nuclear fraction is discussed.     

BIOSYNTHESIS OF RIBONUCLEIC ACID IN RAT BRAIN SLICES

The incorporation of uridine into RNA in brain slices was studied. Optimal conditions for uridine incorporation were determined. The characteristics of the product suggest that de novo DNA-directcd synthesis of fairly high molecular weight material takes place. Incorporation into RNA of several areas of brain was studied. The incorporation was also studied as a function of the age of the animal. Finally, an apparent correlation was observed between the decrease in uridine incorporation with age and the increase of the enzyme uridine nucleosidase which hydrolyses uridine to uracil, a material which cannot be incorporated into RNA.     

METABOLISM OF RIBOSOMAL PRECURSOR RIBONUCLEIC ACID IN KIDNEY

The labile precursors of ribosomal RNA in mouse kidney are preserved when nuclei rapidly isolated after sieving through multiple screens are swollen and cleansed in the presence of an RNase inhibitor before digestion with DNase and phenol extraction. The kinetics of nucleolar labeling analyzed on polyacrylamide gels show that 36S RNA is the major intermediate product in the catabolism of the original 45S RNA precursor to 32S RNA, from which 28S RNA is derived. Each kidney nucleus contains about 200–600 molecules of 45S RNA; the turnover time of the 45S pool is about 3 ± 2 min. Compared with HeLa cells, kidney nuclei have a different major intermediate product and a much smaller and more rapidly turning-over pool of ribosomal precursor RNA.