One cell from the liver of a control (sort of) for the Gclc ko mouse (that is that it had one wild type allele for the gene for turning on albumen synthesis (WC/ii  ) which rescues the would-be Gclc-/- mouse from death. This hepatocyte is not really remarkable to me but it did have this great tangential section of the hepatocyte nucleus and the nuclear pores were just scattered all over the area sectioned making the uniformity of the condensed chromatin areas surrounding the pores very visibly7 consistent.  It was so nicely consistent that I decided to try an afix a casual, but hopefully useful, measurement to that exclusion zone, at least in the this particular animal model.

So, that distance (based on the 27nm size of a mammalian ribosome turns out to be the following:  mean distance, 58nm.  From the original scanned image 58 nm+/- 16nm. That makes the space around the nuclear pore sort of a ring, of about 55 nm. Black line composit measurements, see bottomright. Red dot = one ribosome,

It is nice to read that something is measurable, that has a dimension in nm that one can put into some kind of life-perspective. I don’t know how accurate those individuals were in giving a distance measurement to their images are, and I am giving each the benefit of the doubt in terms of attempting to be accurate) i guess they are quite accurate. I know for myself, using three or four different electron microscopes, and various different paper products, films, dryers, etc, that there is shrinkage, stretching, and all sorts of changes that have occurred over time (even 35 years) that affected the dimensions of objects on the micrographs, even though by a very small amount.

I am using 27nm as an internal control for magnification+enlargement of images. In this case, the 27nm red dot, in all images has been derived from about a dozen ribosomes, and labeled as such. The green dots are about 18-20nm in diameter in these micrographs, and strung as if “beads on a string” (not the beads on a string of nucleosomal DNA, but production of preRNA in the nucleus?) but is it not strange that there is a repeated motif – that is- beads on a string- showing up a lot, both in the nucleoplasm, the nucleolus and the cytoplasm (ribosomes on mRNA) as a kind of “convenient” little pattern.  And who knows where else this pattern might be found). So the green dots show this motif, in a parallel-linear pattern, in the condensed (or closed compartment) chromatin. Both of these examples come from Gclc ko mouse hepatocytes.  Red dots are ribosomes.

I have not really found the equivalent of the electron dense particles in the RER of Gclc mice (posted yesterday) for conditional liver ko animals rescued with NAC at 60d. I have also not found an equivalent (googling and other pathology type searches) for the iron spicules found within those partially RER vesicles.   However I did come close to finding a match for the vesicles within vesicles (though these cells were in vitro, not liver, and not in vivo). The partial publication URL which shows iron particles (USPIO nanoparticles at iron concentrations of 50 μg/ml)(link here) which are NOT LIKE the spicules found in the mice I am looking at, but does show something like the vesicular changes, that is, the vesicle-within a vesicle configuration.

I have pseudocolored the vesicles to compare orange, and on the left is the image from the Gclc conditional ko rescued with NAC, and on the right is the cell line (Canine ADSCs or canine adipose derived stem cells) treated by these researchers with iron.  Red arrows point to the vesicles within vesicles, black arrow from their micrograph point to nanoparticles of iron.  Heavy metal (maybe iron in left hand micrograph) looks like a fuzzy black iron filing mass pulled by a magnet into a glump, very different from the iron in the micrograph on the right. It is critical to point out that the vesicles are pretty much ribosome free, while those RER bound objects with electron dense (presumptive iron) have ribosomes, at least over part of their surface.

Here is another example of metal (in this case aluminum) in tissue culture forming dense lysosomal bodies.

The four (maybe there are more) things that I noticed about the GCLC conditional KO mouse hepatocytes were 1: vesicles within vesicles, 2: lucent areas of mitochondiral matrix adjacent to quite well organized stacked cristi, 3: iron-like staining spicular deposits in dilated RER of hepatocytes, and 4: there is a linked-look between the ballooned areas of the ER (both smooth and rough).

These mice have survived the KO of GCLC, for 60 days, because they were rescued with N-acetyl-cysteine (NAC). NAC reportedly helps replenish the intracellular glutathione supply and can lessen hepatocyte oxidative damage occuring with the knock out of glutamate cysteine ligase catalytic subunit (Gclc), explained HERE. NAC affords protection against the oxidative stress, DNA damage and cancer development and cell death, thus rescues a genetically determined depletion of hepatocyte glutathione. I examined a large series of mouse tissues from these animals and their appropriate controls and while some of these data were published, there was much that never made it into the literature. Thus…. these posts, hoping that someone can be helped by these electron micrographic images of the unrescued, the controls and the NAC-saved mice.

So the micrographs below have RER (albeit the ribosomes are really not that nicely arranged in spirals or grids) and the transition from RER to ER is not consistent with typical RER to SER transitions (that is; both small and large vesicles, empty and full of electron dense filaments; those with center vesicles and those without (almost like continuous segmented mini-intussuceptions, are kind of a large mix of different types, sizes, filled and empty vesicles. These vesicles begin to develope before D60 after NAC rescue, and increase with increasing time.

negative block and animal and treatment: 17902_74138_706_wcii_nac_60d. Size of a ribosome (at 27 nm) was used to calculate the bar markers (smallest black dot below. Inset from image on left is bounded with a white box, enlarge just around 2x on the right). Black arrow points to vesicle-within-a-vesicle phenomenon which is totally everywhere; double headed black arrow points to two types of mitochondrial inner membrane organization, one part points to areas which are quite electron-lucent, and one points to cristi that are highly layered and flattened. White arrow points to filamentous electron dense (i would like to say “iron filings” – like) inclusions and the grey arrow points to a telescoping point between two rounded contiguous vesicles with filamentous inclusions.  Occasional ribosomes are seen on the bounding membranes of these vesicles, and as in this case, they look like peristaltic connections, dipping one end into the next rounded vesicle as if there were strictures, or boluses passing through a long undulating series of tubes. Thee are a few coiled ribosomes on an mRNA through out the cytoplasm, and not pictured some microbodies (dense bodies with uricase crystals). Image to right shows the open-closed RER, SER, ER profiles show internally.

Electron micrograph: liver: Alb w/w Gclc i/i D120 NAC. So here is a perfect example of not having the right data written down while doing collaborative ultrastructure on a project for a colleague.  So after so many years I have to assume that this is a control liver receiving NAC for 120 days after birth, for a conditional Gclc ko mouse.

Liver looks really nice, WNL in my opinion, nice RER, nice mitochondira, one part of a lipid droplet visible. I partially hid two pen marks on this micrograph (using the bandaid tool in photoshop) which you can still see, but this did not change any data. Contrast enhanced as well to even up the tone of the liver histology.  liver, neg 17909, block 74119, anm#702, alb w/w Gclc i/i NAC 120 days post partum. 10000x orig mag. No profile of nucleus in this image.