NUCLEOLAR AND BIOCHEMICAL CHANGES DURING UNBALANCED GROWTH OF TETRAHYMENA PYRIFORMIS

Numerous nucleoli can be observed in the macronucleus of the logarithmically growing ciliated protozoan Tetrahymena pyriformis; at late log phase the nucleoli aggregate and fuse. In stationary phase this fusion process continues, leaving a very few large vacuolated nuclear fusion bodies in the nucleus. When these stationary phase cells are placed into fresh enriched proteose peptone medium, the large fusion bodies begin to disaggregate during the 2.5-hour lag phase before cell division is initiated. By 3 to 6 hours after inoculation the appearance of the nucleoli in many cells returns to what it was in logarithmic cells. In view of the possible role of nucleoli in ribosome synthesis, attempts were made to correlate the morphological changes to changes in RNA and protein metabolism. The beginning of an increased RNA synthesis was concomitant with the beginning of disaggregation of the large fusion bodies into nucleoli, which was noticed in some cells by 1 hour after the return to fresh enriched proteose peptone medium. Increased protein synthesis then followed the increased RNA synthesis by 1 hour. The supply of RNA precursors (essential pyrimidines) were removed from cultures which were grown on a chemically defined synthetic medium, in order to study the relation between nucleolar fusion and synthesis of RNA and protein. Pyrimidine deprivation drastically curtailed RNA and protein synthesis, but did not cause fusion of nucleoli. When pyrimidines were added back to this culture medium, RNA synthesis was immediately stimulated and again preceded an increased protein synthesis by 1 hour. These studies suggest the involvement of unfused nucleoli in RNA and protein synthesis and demonstrate the extreme plasticity of nucleoli with respect to changes in their environment.     

在生长、功能和萎退各期,大鼠乳腺磷酸酶的研究

前言虽然乳腺在生长、功能和萎退过程中形态上的变化是很大的(姚曾序、顾国彦,1957),可是和形态变化相伴随的化学和生物化学变化却研究得少。关于碱性磷酸酶(以下简称 AKP)方面,Kay(1925),Folley 和 Kay(1935)最早证明牛、羊和豚鼠乳腺含有相当量的这种酶。以后 Folley 和 Greenbaum(1947)进一步用生化方法研究了大鼠在怀     

SECRETION OF ACID HYDROLASES AND ITS INTRACELLULAR SOURCE IN TETRAHYMENA PYRIFORMIS

Axenic Tetrahymena pyriformis, syngen 1, mating type II cells were grown in Cox's defined medium. When washed and transferred into nonnutrient dilute salt solution or resuspended in the defined medium, the intact cells secrete acid hydrolases into the medium. Cells starving in the salt solution release in 5 hr about two-thirds of their β-glucosidase, β-N-acetylglucosaminidase, α-glucosidase, and amylase activities, about one-third of their deoxyribonuclease and phosphatase activities, smaller amounts of ribonuclease, and only a negligible fraction of their proteinase activity and protein content. During this period there is practically no change in the enzyme activities (except for a sudden increase of ribonuclease activity) and protein content of cells and medium together. Cells resuspended in the nutrient medium secrete enzymes as do the starved cells, but replace this loss, so that there is a continuous increase of the activities in the total system. According to isopycnic centrifugation experiments performed in sucrose gradients, the source of the hydrolases is a special population of lysosomes which disappear from the cells during starvation. This population equilibrates in the high density region of the gradients and contains the various acid hydrolases in about the proportion in which these enzymes appear in the medium.     

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.     

小麦雌配子体发育过程中酸性磷酸酶活性定位

在小麦(Triticum aestivum L.)雌配子体发育过程中,胚囊周围邻近的珠心细胞退化降解,并出现很高的酸性磷酸酶反应,特别是合点部分最强。电镜细胞化学定位也表明退化珠心细胞质中有强烈的酸性磷酸酶活力,它们存在于多层环状的胞质结构中,而远离胚囊的非退化珠心细胞中无上述结构,酸性磷酸酶活性仅出现于液泡中。认为珠心细胞的退化是一种自溶现象。从功能大孢子至七细胞胚囊期,胚囊内部胞质酸性磷酸酶活性很低,合点与珠孔两端的反应强度无明显区別。后期成熟胚囊阶段,反足细胞中出现强烈酸性磷酸酶活性,中央细胞次之,而助细胞及卵细胞中很弱。     

THE FATE OF MITOCHONDRIA DURING AGING IN TETRAHYMENA PYRIFORMIS

During the growth cycle of Tetrahymena pyriformis the mitochondria undergo changes in position, number, and structure. Ciliates in the logarithmic growth phase possess elongated mitochondria which are aligned along the plasma membrane and are closely associated with the kinetosomes and kinetodesmata. Mitochondria appear to divide across the long axis at this time, resulting in two or more products. Throughout this phase of growth mitochondrial divisions keep pace with cytokinesis so that the population of mitochondria remains at essentially the minimal level. As the ciliates enter the stationary growth phase the mitochondria increase in number, become oval to spherical in shape, and some migrate into the cytoplasm. Intramitochondrial masses of various configurations appear at this time. Some of the mitochondria lying in the cytoplasm become incorporated into vacuoles. Within these vacuoles either a single mitochondrion appears or several mitochondria may be seen along with other cytoplasmic structures. Later in the stationary growth phase the contained mitochondria are dense and the tubules are more compact than normal. Various stages in disorganization of the mitochondria are observed in a single large vacuole. Cytochemical tests reveal the presence of acid phosphatase, suggesting that hydrolysis of the vacuolar contents occurs. Lipid droplets increase in number during the middle and late stationary phase of growth. These events are interpreted as being associated with the normal process of aging in T. pyriformis.     

小麦受精过程中酸性磷酸酶的超微细胞化学定位

小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）受精前成熟胚囊,除胚囊中央细胞的合点端细胞质中有酸性磷酸酶外,其余部位均未发现酸性磷酸酶。受精时期,以下部位存在酸性磷酸酶活性：卵细胞的细胞核内一部分染色质和细胞质中大部分线粒体;精、卵核融合时两核的核周腔内;退化助细胞合点端细胞质和一些液泡内;进入雌性细胞中的两个精核;胚囊各成员细胞的细胞壁及胚囊周围珠心细胞的细胞壁。二细胞原胚中未见有酸性磷酸酶。早期胚乳游离核染色质上有酸性磷酸酶。小麦受精过程酸性磷酸酶的分布特点可能与卵细胞生理状态的变化和细胞质中线粒体的改组、助细胞的退化、精核的生理状态以及精核与卵核的核膜融合等有关。     

SYNTHESIS OF DEOXYRIBONUCLEIC ACID BY MICRO- AND MACRONUCLEI OF TETRAHYMENA PYRIFORMIS

Evidence as to the times of DNA synthesis in micronucleate Tetrahymena pyriformis (mating type II, variety 1) has been obtained by briefly exposing individuals of different ages to tritiated thymidine, returning them to non-radioactive medium, fixing at division, and preparing autoradiographs. A variable length of interphase, ranging from a few minutes to about 2 hours, has been found to precede the initiation of macronuclear DNA synthesis. Once begun, however, the period of synthesis appears to be similar in all cells, regardless of generation time, and has been estimated at 1 to 1½ hours. Under the conditions of these experiments, the time elapsing between the end of synthesis and subsequent division into daughter cells ranges from approximately 1½ to 2½ hours in generation times long enough to allow such variability. Division of the micronucleus occurs shortly before the cell begins to divide; its DNA synthesis starts immediately and continues after cell division for a total period estimated at about an hour.     

ULTRASTRUCTURE OF MUCOCYSTS AND PELLICLE OF TETRAHYMENA PYRIFORMIS

Tetrahymena pyriformis GL was fixed with glutaraldehyde and/or OsO₄ for a study of cytoplasmic ultrastructure. Many small vacuoles 0.05 to 0.5 µ in diameter were found to contain each a dense particle enveloped by a limiting membrane. This membrane is continuous with the membrane of the vacuole. The particles are irregular in shape and size, but similar to the mucocysts in the appearance of the matrix. It is suggested that they are the first morphologically distinguishable stages in the development of mucocysts. In the course of this development, amorphous material becomes crystalline with a longitudinal period of 150 A and a lateral period of 100 A. The mature mucocysts are rather uniform in size and have a spheroidal shape. During discharge, the crystalline pattern disappears and the mucocysts assume a spherical configuration. The inner limiting membrane of a mucocyst seems to disintegrate during the process of discharge while the outer membrane becomes continuous with the outermost pellicular membrane; the inner pellicular membrane is continuous with the cytoplasmic membrane. Rows of few to 15 or more microtubules were found either between the cytoplasmic membrane and the ectoplasmic layer (longitudinal fibrils) or underneath the ectoplasmic layer (transverse fibrils). The outer and inner pellicular membranes are uniformly spaced and connected by "cross-bridges." Details of these structures are described.     

水稻叶片酸性磷酸酯酶活性及其部分特性

从水稻叶片部分纯化了水解磷酸烯醇式丙酮酸的磷酸酯酶,其Ｋｍ（ＰＥＰ）为０．１ｍｍｏｌ／Ｌ,最适ＰＨ５．３．在偏酸性ＰＨ条件下（ＰＨ４．０～７．２）稳定,对热亦较稳定．酶活性受Ｐｉ强烈抑制．它对其底物要求不专一,能水解多种含磷酯键的化合物．表明它是一种非专一性的酸性磷酸酯酶。各种含磷酯键的代谢物对酶活性起竞争性抑制作用,且表现出叠加性．Ｃｕ（２＋）、Ｚｎ（２＋）和Ｆｅ（２＋）抑制酶活性,Ｍｇ（２＋）、Ｍｎ（２＋）、Ｃａ（２＋）、Ｃｏ（２＋）和ＥＤＴＡ无影响．     

SIMULTANEOUS SYNTHESIS OF HISTONE AND DNA IN SYNCHRONOUSLY DIVIDING TETRAHYMENA PYRIFORMIS

Histone and DNA syntheses have been studied in synchronously dividing Tetrahymena pyriformis GL. During the heat treatment necessary to synchronize cultures of this amicronucleate protozoan, the DNA content of the already polyploid macronucleus increases. When the cells begin synchronous division, their DNA content is reduced in a stepwise process which is closely paralleled by reduction of macronuclear histone content. During cell division, the contents of DNA and histone decrease by slightly more than twofold, and in the subsequent S phase, DNA and histone increase simultaneously to 85% of the values expected if all chromosomes were to double. The first step in the process of reduction of DNA and histone contents is their decrease in excess of twofold, and this is accomplished by removal of extrusion bodies from the nuclei of dividing cells. The second step is a mechanism which allows, in effect, only 70% of the chromatin in the average nucleus to duplicate. Such partial duplication suggests that both histone and DNA syntheses in synchronous Tetrahymena depend upon a regulatory mechanism, the mediating elements of which are localized in only certain chromosomes.     

菓蝇变态过程中磷酸酶的性质和变化

昆虫变态是一个复杂的生物学过程。经过这个过程,完全变态的昆虫从幼虫转变为成虫。由于幼虫和成虫的生活方式截然不同,在变态时发生了幼虫组织的退化和破坏以及成虫组织的新生和分化。这也就是一般所谓组织分解(Histolysis)和组织发生(Histogenesis)两个现象。关于变态的生物学和生理学,人们已积累了相当数量的知     

IDENTIFICATION OF LECTINS IN THE KINETIDS OF TETRAHYMENA PYRIFORMIS

Previously we described lectin-like molecules in the ciliate Tetrahymena pyriformis; by application of synthetic neoglycoconjugates it is now shown that T. pyriformis contains considerable amounts of both a β-d-glucose- and a lactose-specific lectin. No evidence for the presence of α-d-mannose-, α-d-galactose- or of α-l-fucose-specific lectins could be obtained. The two lectins, identified in T. pyriformis, are associated with the kinetids. During cell division the lectins disappear or become masked in the fission furrow. Therefore, we assume that these lectins are involved in the organization of the distribution pattern of the kinetids during cell division perhaps due to lectin—glycoprotein interactions.     

大豆子叶内酸性磷酸酶活性的超微结构定位

开花后３５～５０ ｄ 期间和萌发早期（播种后４～８ ｄ）的大豆（Ｇｌｙｃｉｎｅｍ ａｘ Ｌ．）种子中,酸性磷酸酶主要分布在子叶细胞中的蛋白体内;在内质网内也检测到酸性磷酸酶活性。此外,在萌发早期的部分子叶细胞的质膜外侧及其细胞壁基质中可见密集的酸性磷酸酶活性;而且在近质膜的胞质中常见到一些富含磷酸铅沉淀的胞质小泡,似与质膜融合