Evaluation of Cell Death in EB V-Transformed Lymphocytes Using Agarose Gel Electrophoresis, Light Microscopy and Electron MicroscopyI. Induction of Classic Apoptosis by the Bile Salt, Sodium Deoxycholate

Cultures of bovine pulmonary endothelial (BPE) cells were exposed to LC70 doses of ricin or abrin (15.5 and 4.5 pM respectively) over a period of up to 40 h. The viability of the cultures (as determined by the neutral red (NR) dye retention assay) declined after 6 h exposure to the toxins. From 15 h onwards, cellular material in toxin exposed cultures became detached from the substra tum of the culture vessels. Hoffman modulation contrast photomicrography showed that this process was due to ricin and abrin exposed cells collapsing into membrane bound vesicles which retained the NR dye, became detached and floated into the medium. These apoptotic-like structural changes were further investi gated by transmission electron microscopy (TEM) and by agarose gel electrophoresis of DNA from control and exposed cultures. Many of the characteristic changes associated with apoptotic cell death were seen using TEM, including heterochromatin condensation at the nuclear periphery, crenulation of the nuclear mem brane and progressive degeneration of residual nuclear and cytoplasmic structures. The plasma membrane of many cells remained intact, and contained nuclear and cytoplasmic debris. Agarose gel electrophoresis of DNA extracted from toxin-treated cells revealed oligonucleosome sized DNA fragments, characteristic of apoptosis, from adherent cells at 7 h and both adherent and floating populations when harvested from 15 h; DNA from unexposed control cells did not show this fragmentation. The identification of apopto sis as being a significant additional mechanism of toxicity following exposure to ricin and abrin holotox ins raises the possibility of developing new therapeutic strategies against poisoning by these phytotoxins.

Download Full-text

Gonadotropin suppression of apoptosis in follicle somatic cells

Zygote ◽

10.1017/s0967199400003282 ◽

1996 ◽

Vol 4 (04) ◽

pp. 299-303 ◽

Cited By ~ 1

Author(s):

G. Galeati ◽

M. Forni ◽

M. Spinaci

Keyword(s):

Cell Death ◽

Granulosa Cells ◽

Gel Electrophoresis ◽

Genomic Dna ◽

Apoptotic Cell ◽

Apoptotic Cell Death ◽

Agarose Gel ◽

Agarose Gel Electrophoresis ◽

Follicular Atresia ◽

Prominent Feature

In each oestrous cycle only a limited number of follicles are selected for ovulation whereas the remaining majority undergo atresia. The earliest and most prominent feature of atresia is the death of granulosa cells. Recent biochemical evidence has demonstrated that granulosa cell death during follicular atresia in swine (Tillyet al., 1992), bovine (Jollyet al., 1994) and rodent (Tillyet al., 1991) ovaries occurs by apoptosis, a process whereby cells die in a controlled manner. A biochemical event considered to be characteristic of apoptotic cell death is the intranucleosomal cleavage of genomic DNA into fragments 180–200 bp in size, which separate into a distinctive ladder-like pattern on agarose gel electrophoresis. Detection of this pattern of oligonucleosomes in DNA provides a marker of apoptotic cell death.

Download Full-text

Luminography – an Alternative Assay for Detection of von Willebrand Factor Multimers

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647016 ◽

1988 ◽

Vol 60 (02) ◽

pp. 133-136 ◽

Cited By ~ 50

Author(s):

R Schneppenheim ◽

H Plendl ◽

U Budde

Keyword(s):

Von Willebrand Factor ◽

Gel Electrophoresis ◽

Detection Methods ◽

Agarose Gel ◽

Agarose Gel Electrophoresis ◽

Luminescence Assay ◽

Von Willebrand ◽

Willebrand Factor

SummaryA luminescence assay was adapted for detection of von Willebrand factor multimers subsequent to SDS-agarose gel electrophoresis and electroblotting onto nitrocellulose. The method is as fast as chromogenic detection methods and appears to be as sensitive as autoradiography without the disadvantages of the latter.

Download Full-text