Apoptosis

A. Cell proliferation and apoptosis

The number of cells in any tissue is mainly regulated by two processes—cell
proliferation and physiological cell death, apoptosis. Both of these processes
are regulated by stimulatory and inhibitory factors that act in solute form (growth
factors and cytokines) or are presented in bound form on the surface of
neighboring cells (see below). Apoptosis is genetically programmed cell death,
which leads to “tidy” breakdown and disposal of cells .Morphologically, apoptosis
is characterized by changes in the cell membrane (with the formation of small
blebs known as “apoptotic bodies”), shrinking of the nucleus, chromatin
condensation, and fragmentation of DNA. Macrophages and other phagocytic
cells recognize apoptotic cells and remove them by phagocytosis without
inflammatory phenomena developing. Cell necrosis (not shown) should be
distinguished from apoptosis. In cell necrosis, cell death is usually due to physical
or chemical damage. Necrosis leads to swelling and bursting of the damaged
cells and often triggers an inflammatory response. The growth of tissue (or, more
precisely, the number of cells) is actually regulated by apoptosis. In addition,
apoptosis allows the elimination of unwanted or superfluous cells—e. g., during
embryonic development or in the immune system. The contraction of the uterus
after birth is also based on apoptosis. Diseased cells are also eliminated by
apoptosis—e. g., tumor cells, virus-infected cells, and cells with irreparably
damaged DNA. An everyday example of this is the peeling of the skin after
sunburn.
B. Regulation of apoptosis
Apoptosis can be triggered by a number of different signals that use various
transmission pathways. Other signaling pathways prevent apoptosis. At the
center of the apoptotic process lies a group of specialized cysteine-containing
aspartate proteinases , known as caspases. These mutually activate one
another, creating an enzyme cascade resembling the cascade involved in blood
coagulation . Other enzymes in this group, known as effector caspases, cleave
cell components after being activated—e. g., laminin in the nuclear membrane
and snRP proteins — or activate special DNases which then fragment the nuclear
DNA.
An important triggerfor apoptosis is known as the Fas system. This is used by
cytotoxic T cells, for example, which eliminate infected cells in this way (top
left).Most of the body’s cells have Fas receptors (CD 95) on their plasma
membrane. If a T cell is activated by contact with an MHC presenting a viral
peptide , binding of its Fas ligands occurs on the target cell’s Fas receptors. Via
the mediator protein FADD (“Fasassociated death domain”), this activates
caspase- 8 inside the cell, setting in motion the apoptotic process. Another
trigger is provided by tumor necrosis factor- α (TNF-α), which acts via a similar
protein (TRADD) and supports the endogenous defense system against tumors by
inducing apoptosis. Caspase-8 activates the effector caspases either directly, or
indirectly by promoting the cytochrome c from mitochondria. Once in the
cytoplasm, cytochrome c binds to and activates the protein Apaf-1 (not shown)
and thus triggers the caspase cascade. Apoptotic signals can also come from the
cell nucleus. If irreparable DNA damage is present, the p53 protein —the
product of a tumor suppressor gene—promotes apoptosis and thus helps
eliminate the defective cell. There are also inhibitory factors that oppose the
signals that activate apoptosis. These include bcl-2 and related proteins. The
genomes of several viruses include genes for this type of protein. The genes are
expressed by the host cell and (to the benefit of the virus) prevent the host cell
from being prematurely eliminated by apoptosis.
18.6.6. Mitochondria Play a Key Role in Apoptosis
In the course of development or in cases of significant cell damage, individual cells within
multicellular organisms undergo programmed cell death, or apoptosis. Mitochondria act as control
centers regulating this process. Although the details have not yet been established, a pore called the
mitochondrial permeability transition pore (mtPTP) forms in damaged mitochondria. This pore
appears to consist of VDAC (the adenine nucleotide translocator) and several other mitochondrial
proteins, including members of a family of proteins (Bcl family) that were initially discovered because
of their role in cancer. One of the most potent activators of apoptosis is cytochrome c. Its presence in
the cytosol activates a cascade of proteolytic enzymes called caspases. These cysteine proteases
(Section 9.1.6) are conserved in evolution, being found in organisms ranging from hydra to human
beings. Cytochrome c, in conjunction with other proteins, initiates the cascade by activating
procaspase 9 to form caspase 9, which then activates other caspases. Activation of the caspase cascade
does not lead to generalized protein destruction. Rather, the caspases have particular targets. For
instance, the proteins that maintain cell structure are destroyed. Another example is the degradation of
a protein that inhibits an enzyme that destroys DNA (caspase-activated DNAse, CAD), freeing CAD
to cleave the genetic material. This cascade of proteolytic enzymes has been called "death by a
thousand tiny cuts."

Programmed Cell Death

At certain stages in the development of a multicellular organism, some cells must die. This well-
regulated process is called is called apoptosis (programmed cell death), as suggested by Kerr in 1972.
The importance of this biological phenomenon was first realized in studies of a tiny worm, the soil
nematode C. elegans (see p. 304). If apoptosis does not occur, developmental failure of the organism
or cancer may result. Apoptosis is regulated in the apoptosis pathway by many proteins, which either
trigger or prevent apoptosis. Apoptosis can be triggered by a variety of stimuli that act either from
outside the cell (extrinsic pathway) or from within the cell (intrinsic pathway). External stimuli may
be irradiation, withdrawal of essential growth factors, or glucocorticoids. An intrinsic stimulus may be
spontaneous damage to the DNA of the cell.
A. Importance of apoptosis
Apoptosis occurs mainly during development. For example, the digits in the developing mammalian
embryo are sculptured by apoptosis (1). The paws (hands in humans) start out as spadelike structures.
The formation of digits requires that cells between them die (here shown as bright green dots on the
left). More staggering is the amount of apoptosis in the developing vertebrate nervous system.
Normally up to half of the nerve cells die soon after they have been formed. In the embryos of mice
that lack an important gene regulating apoptosis (caspase 9, see below), neurons proliferate
excessively and the brain protrudes above the face (2). (Illustration in 1 modified from Alberts et al.,
2002; in 2 from Gilbert, 2003, according to Kaida et al., 1998.)
B. Cellular events in apoptosis
The first visible signs of apoptosis are condensation of chromatin and shrinking of the cell. The cell
membrane shrivels (membrane blebbing), and the cell begins to disintegrate (nuclear segmentation,
DNA fragmentation). An apoptotic body of cell remnants forms, which eventually dissolves by a
process called lysis. (Figure modified from Dr. A. J. Cann, Microbiology, Leicester University,
displayed at Google Images, 22 March, 2005.)
C. Regulation of apoptosis
Specialized cysteine-containing aspartate proteinases, called caspases, play a central role. They
activate or inactivate each other in a defined sequence. Binding of a ligand, Fas, of a cytotoxic T cell
(see section on immune system) to the Fas receptor (also called CD95) activates an intracellular
adaptor protein, FADD (Fas-associated death domain). This binds to and activates procaspase 8 into
active caspase 8. Caspase 8 causes release of cytochrome c in mitochondria (see p. 130) and activates
several different effector caspases. Downstream of caspase 8, two pathways exist. In type I cells (in
thymocytes and fibroblasts), caspase 8 directly activates caspase 3. In type II cells, such as
hepatocytes, caspase 8 cleaves Bid, a member of the Bcl-2 family. The mouse and human genomes
contain 13 caspase genes (1–12 and 14; see table in the appendix). Human caspases 3 and 6–10 are
involved in apoptosis, the others in inflammation (Nagata, 2005). Caspase 8 also serves as a selective
signal transducer for nuclear factor KB (NFkB) during the early genetic response to an antigen (Su et
al., 2005). Other regulators of apoptosis are members of the Bcl-2 family (the name Bcl is derived
from B-cell lymphoma, a human malignant tumor originating from B-lymphocytes and caused by
mutations in this gene). (Figure from Koolman & Rِhm, 2005.)