Three-dimensional reconstruction of organelles in Euglena gracilis Z. II. Qualitative and quantitative changes of chloroplasts and mitochondrial reticulum in synchronous cultures during bleaching

1980 ◽  
Vol 46 (1) ◽  
pp. 313-340
Author(s):  
M. Pellegrini

By ultrathin serial sectioning, morphological and volumetric changes of the plastidome and chondriome have been observed in Euglena gracilis Z during bleaching in darkness with addition of sodium acetate to the culture medium. In order not to introduce any modification to the synchronization pattern during bleaching, green cells were previously grown photoautotrophically on Cramer & Myers medium under continuous illumination and synchronized by temperature cycles and (2) of sodium acetate and darkness on the plastidome and chondriome of photoautotrophic cells synchronized by light-dark cycles as described previously. In photoautotrophic cells, the plastidome, consisting of about ten diskoidal chloroplasts, occupies 15% of the cell volume. The chondriome, in the form of one single giant mitochondrion branched throughout the cell, represents 6% of the cell volume. The synchronization by temperature cycles in continuous illumination does not change the morphology and volume of these organelles. However, pyrenoids disappear. In photoheterotrophic culture with sodium acetate added, the plastidome fine structure does not vary but its volume decreases by 19–25%. At that time, the plastidome thus occupies 12–13% of the cell volume. Sodium acetate provokes, on the contrary, hypertrophy of the delicate threads of the mitochondrial reticulum which appears as a network with narrow meshes around other organelles. The chondriome thus comes to occupy 10–11% of the cell volume. In heterotrophic cells, the combined effects of sodium acetate and darkness emphasize the regression of the plastidome while the chondriome appears as a fenestrated parietal shell occupying 15–16% of the cell volume. Maximal hypertrophy is obtained in 24 h. Total dedifferentiation of chloroplasts requires 6–9 successive generations in heterotropic conditions. These results are discussed in relation to numerous light-microscopic and ultrastructural observations. It has been demonstrated, as in photoautotrophic Euglena cells synchronized by light-dark cycles, that the plastidome of heterotrophic cells consists of about ten organelles, whereas the chondriome contains one single giant mitochondrion. Contrary to the opinion that the variations of the plastidome and chondriome are reciprocally related, it is proved here that dedifferentiation of chloroplasts and hypertrophy of the chondriome occur at different rates, and may be independent of one another.

1980 ◽  
Vol 43 (1) ◽  
pp. 137-166
Author(s):  
M. Pellegrini

Ultrastructural changes of chloroplasts and mitochondria have been observed in synchronously growing cells of Euglena gracilis Z, under photoautotrophic conditions. Application of the serial section technique allows estimation of the number and volume of these organelles. Several 3-dimensional reconstructions reveal their shape and distribution throughout the cell cycle. In young cells 10 separate diskoid or branched chloroplasts are found. They show the typical lamellar structure of euglenoid chloroplasts. During the growth phase (light period), they enlarge and their volume doubles. Some of them branch out, so that 20 lobes are formed. Thylakoids grow longer without change in number. The pyrenoid persists only during the first half of this period. During the cell division phase (dark period), branched chloroplasts divide along 2 planes which are perpendicular to each other and perpendicular to the thylakoid plane. All thylakoids are cut and their number does not change in the daughter chloroplasts. The plastidome volume constitutes 15–18% of the total cell volume over the entire life cycle. One of the most significant observations in this report is the presence of a single permanent mitochondrial reticulum during the whole cell cycle. This giant mitochondrion consists of an extremely branched network with delicate threads (0.4-0.6 micrometer thick) surrounding the chloroplasts, nucleus and reservoir. It extends throughout the cell. During the growth phase, it becomes gradually longer and doubles in volume. The degree of branching increases but the thickness of the threads remains constant. During the division phase, the mitochondrial elements appear more restricted (0.4 micrometer thick) and the reticulum becomes progressively partitioned into 2 daughter networks. At any time of the cell cycle, the chondriome volume is about 6% of the total cell volume. These results are discussed in comparison with numerous relevant papers on light and electron microscopy of animal and plant cells. They suggest that the descriptions of several authors on the plastidial cycle and the mitochondrial cycle in Euglena, both said to be characterized by alternate reticulate and fragmentary states, arise in part from questionable interpretation of random sections. It is evident that the form and distribution of organelles can be determined more precisely by serial sectioning.


Author(s):  
Jerome J. Paulin

Within the past decade it has become apparent that HVEM offers the biologist a means to explore the three-dimensional structure of cells and/or organelles. Stereo-imaging of thick sections (e.g. 0.25-10 μm) not only reveals anatomical features of cellular components, but also reduces errors of interpretation associated with overlap of structures seen in thick sections. Concomitant with stereo-imaging techniques conventional serial Sectioning methods developed with thin sections have been adopted to serial thick sections (≥ 0.25 μm). Three-dimensional reconstructions of the chondriome of several species of trypanosomatid flagellates have been made from tracings of mitochondrial profiles on cellulose acetate sheets. The sheets are flooded with acetone, gluing them together, and the model sawed from the composite and redrawn.The extensive mitochondrial reticulum can be seen in consecutive thick sections of (0.25 μm thick) Crithidia fasciculata (Figs. 1-2). Profiles of the mitochondrion are distinguishable from the anterior apex of the cell (small arrow, Fig. 1) to the posterior pole (small arrow, Fig. 2).


Author(s):  
Tomoko Ehara ◽  
Shuji Sumida ◽  
Tetsuaki Osafune ◽  
Eiji Hase

As shown previously, Euglena cells grown in Hutner’s medium in the dark without agitation accumulate wax as well as paramylum, and contain proplastids showing no internal structure except for a single prothylakoid existing close to the envelope. When the cells are transferred to an inorganic medium containing ammonium salt and the cell suspension is aerated in the dark, the wax was oxidatively metabolized, providing carbon materials and energy 23 for some dark processes of plastid development. Under these conditions, pyrenoid-like structures (called “pro-pyrenoids”) are formed at the sites adjacent to the prolamel larbodies (PLB) localized in the peripheral region of the proplastid. The single prothylakoid becomes paired with a newly formed prothylakoid, and a part of the paired prothylakoids is extended, with foldings, in to the “propyrenoid”. In this study, we observed a concentration of RuBisCO in the “propyrenoid” of Euglena gracilis strain Z using immunoelectron microscopy.


Author(s):  
L Epprecht ◽  
L Qingsong ◽  
N Stenz ◽  
S Hashimi ◽  
T Linder

Abstract Objective Ventilation of the middle ear and mastoid air cells is believed to play an important role in the pathogenesis of chronic ear disease. Traditionally, ventilation is assessed by computed tomography. However, this exposes patients to cumulative radiation injury. In cases with a perforation in the tympanic membrane, tympanometry potentially presents a non-invasive alternative to measure the ventilated middle-ear and mastoid air cell volume. This study hypothesised that total tympanometry volume correlates with ventilated middle-ear and mastoid air cell volume. Method Total tympanometry volume was compared with ventilated middle-ear and mastoid air cell volume on computed tomography scans in 20 tympanic membrane perforations. Results There was a high correlation between tympanometry and computed tomography volumes (r = 0.78; p < 0.001). A tympanometry volume more than 2 ml predicted good ventilation on computed tomography. Conclusion These results may help reduce the need for pre-operative computed tomography in uncomplicated cases with tympanic membrane perforations.


Microbiology ◽  
1988 ◽  
Vol 134 (1) ◽  
pp. 61-66
Author(s):  
K. HOSOTANI ◽  
T. OHKOCHI ◽  
H. INUI ◽  
A. YOKOTA ◽  
Y. NAKANO ◽  
...  

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