Category Archives: Golgi, vesicles and stress

golgi apparatus electron microscopy post translational modification stress protein accumulation protein transport

Variety: the whole spectrum of lysosomes/inclusions of E2 in an alveolar macrophage

Electron microgaphs of lung, Golgi, vesicles and stress, Legacy Perfluorocarbons, The cell: images, Ultimate order, the cell

Variety: the whole spectrum of lysosomes/inclusions of E2 in this micrograph of an alveolar macrophage (at least the whole variety I have seen so far). It includes lysosomes (LE/LY/PFC) which probably could be classified all the way from golgi vesicles to late endosomes since it seems that PFCs in general are able to work their way into the whole ER system (though to date I have not seen anything that looks like a PFC droplet in any ER which has ribosomes….  still looking). There are droplets in a “hollow” meaning no lysosomal enzymes yet, on the right but still having E2 droplets with the coating they must pick up from somewhere (maybe alveolar surfactant in the alveolar space?, being lipid and E2 being slightly lipophilic), to the dense small lysosomes with many E2 droplets on the left, to a single E2 droplet on the lower left, and a dozen droplets which look like they are hanging out in the cytosol with no membrane boundaries, to bottom center structure which looks like two E2 droplets in a lysosome with a lamellar type surfactant inclusion.

There is no paucity of free ribosomes in this cropped image, and size can be inferred from the approximate 27nm diameter of a ribosome (red dot), to the larger droplet size (bar=270nm).

MVB/LE/PFC bodies within alveolar macrophages of liquid-breathed mice

Electron microgaphs of lung, Environment and contaminants, Golgi, vesicles and stress, Legacy Perfluorocarbons, The cell: images, Toxi-city, Ultimate order, the cell

In this case the liquid which the mouse breathed for 3 hours and allowed to recover for 17 days was E2.  Thank goodness for good record keeping otherwise I would have forgotten what E2 stood for. Here is what the notations say (from back when Leland Clark Jr. was keeping tabs on all documents and publications.  E2 (made by Du Pont) O2 solubility ml/100ml at 25oC 55.7, vapor pressure 56mmHg at 37.5oC, viscosity 0.6 at 25oC, boiling point at 104.4oC, density gm/ml 1.656 at 25oC, surface tension 12.9 dynes/cm at 25oC.E2 liquid for breathing perfluorochemical molecular
There were a bunch of MVB/LE/PFC organelles in these macrophages that I began to look more closely at the parallel leaflet patterns that were showing up at higher magnification.

Altogether there were 29 mice that breathed E2, it much have been a satisfactory fluid or there would not be a dozen or more mice that survived from 5 days to 449 days after 3 hr of breathing E2.

The electron micrograph below was one such body, I don’t think it could clearly be labeled a multivesicular body or a late endosome, but there are E2 droplets involved in its organization.  I don’t believe there is any outer nuclear membrane association, but there are membranes with different distances apart which are clearly organized (actually a parallel disorganization – if such a thing can (and it does visually here) exist).  So I sensed that the distance between the leaflets of these membranes was different within the MVB/LE/PFC thingie and in the membranes arranged just outside the MVB/LE/PFC structure and they were different. The latter look to be about separated at 22nm n=28, x=0.053+0.0014  (mm, on the micrograph translated to close to 22nm in actuality) , and I also measured couple of dozen distances between parallel pairs within the MVB/LE/PFC structure itself and it was closer to 28nm (n=31). When the long string of measurements were subjected to a t-test (remember this is an N of ONE macrophage (there will be others) so it is not really saying much) there was an highly significant (p=0.0001) difference between the mean and SEM of the two (color coded) groups. Pictures below show how the lines and data shaped up. White dotted line outlines the two areas sampled for leaflet separation.

Figure below that shows actual sites of measurement and place of measurement for each group.

liquid breathing electron microscopy mouse lung E2

liquid breathing electron microscopy mouse lung E2In another macrophage –another micrograph — another block same E2 treatment and duration, a similar type of inclusion is seen, but this one did not display the linear qualities of the MVB/LE/PFC as above.  There are two droplets of presumptive E2 in this image, they are middle right and upper left.  Whether the lucent area in this inclusion is E2 is anybody’s guess.

Electron micrographs of golgi and golgi stress

Golgi, vesicles and stress, Science and art cover illustrations, The cell: images, Ultimate order, the cell

Cover submission from the early 2000s for a published manuscript by Shull Miller and Prasad where golgi, golgi ion transport, and golgi membranes (yep, i am not capitalizing the word Golgi any more, with all due respect to Dr. Golgi, this is no longer about him, it is about the organelle and he has had plenty of fame already).

This cover submission was about flattened golgi, cis and trans faces nicely curved, as opposed to dilation and collection of protein contents.  The cover was never executed, but it is a fun graphic, none-the-less.

Photoshop was used to change color gradients of different electron-densities of the membranes and contents. Cytoplasm here primarily, only a portion of other organelles.

There is an obvious texture to the area associated with the dilated ER, how fun it will be (maybe it will happen) to return to those micrographs and hunt for a patterning in the protein aggregates.