Professional Documents
Culture Documents
Cells in Tissues
Bundlethe more are used to describe how cells function as a unit within organ
books you buy, systems, coordinating activities and communicating with one
the greater your another. The endocrine system of insects affects molting and
discount! metamorphosis, and specialized cells are also important in each
of these processes within that organ system. The experiments
that were devised to determine the role of hormones in insect
THE CONTENT
molting and metamorphosis are described. Finally, stem cells
Energy Physics are healthy components of several different systems in animal
Engineering bodies and are described in relation to a disruption in function.
Biotechnology In this breakdown of function, cancer cells, in contrast to stem
Biology cells, can abnormally affect cell cycle regulation.
Mathematics
Christopher J. Paradiseis professor of biology and environ-
Chemistry
mental studies at Davidson College. He teaches introductory
biology, ecology, entomology, and topical seminars on ecotoxi-
THE TERMS cology and renewable natural resources. He also occasionally
Perpetual access leads a study abroad program in India. His research evaluates
for a one time fee anthropogenic factors that influence insect biodiversity at a
No subscriptions or variety of scales. His current research interests include effects of
access fees land use patterns on pollinator communities in parks.
Unlimited
A. Malcolm Campbellteaches biology at Davidson College,
concurrent usage
NC. He received national and international education awards:
Downloadable PDFs Genetics Society of America (2013); American Association for the
Free MARC records Advancement of Science (2012); and American Society for Cell
Biology (2006). He was the founding co-editor in chief of CBE Life
For further information,
Sciences Education; founding director of Genome Consortium
a free trial, or to order,
contact: for Active Teaching (GCAT); and member of the American Soci- Christopher J. Paradise
sales@momentumpress.net ety for Cell Biology governing council (20122014).
A. Malcolm Campbell
Cells in Tissues
Cells in Tissues
10 9 8 7 6 5 4 3 2 1
Keywords
multicellular organisms, undifferentiated cells, stem cells, tissues,
muscle cells, neurons, alimentary canal, information, ingestion, diges-
tion, assimilation, homestasis, stomach, small intestines, epithelial
cells, epithelium, smooth muscle, gastric pits, parietal cells, histamine,
proton pump inhibitor, tubulovesicles, pepsinogen, pepsin, peristalsis,
endocrine system, hormones, exoskeleton, molting, epidermis, hemo-
lymph, corpus allatum, metamorphosis, protocerebrum, prothoracic
gland, neurosecretory cells, stem cells, cancer cells, tumors, multipo-
tent, tumor suppressors, mitogen, central nervous system, peripheral
nervous system, carcinoma
Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 The Digestive System Breaks Down and Absorbs
Nutrients from Ingested Food............................................1
Chapter 2 Populations of Endocrine Cells in Animals Affect the
Whole Organism.............................................................17
Chapter 3 Similarities and Differences Between Stem Cells and
Cancer Cells.....................................................................27
Ethical, Legal, Social Implications: Decisions
Aboutto Whom and When Medical Interventions
are Given......................................................................38
Conclusion............................................................................................43
Glossary................................................................................................45
Index....................................................................................................49
Preface
This book about cells and how they are part of tissues and organisms
is part of a thirty book series that collectively surveys all of the major
themes in biology. Rather than just present information as a collection
of facts, the reader is treated more like a scientist, which means the
data behind the major themes are presented. Reading any of the thirty
books by Paradise and Campbell provides readers with biological con-
text and comprehensive perspective so that readers can learn important
information from a single book with the potential to see how the major
themes span all size scales: molecular, cellular, organismal, population
and ecologic systems. The major themes of biology encapsulate the
entire discipline: information, evolution, cells, homeostasis and emer-
gent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
In this book, readers will learn about cells in tissues and some of the sup-
porting evidence behind our understanding. The historic and more recent
experiments and data will be explored. Instead of believing or simply
accepting information, readers of this book will learn about the science
behind cells in the context of tissues the way professional scientists do
with experimentation and data analysis. In short, data are put back into the
teaching of biological sciences.
Readers of this book who wish to see the textbook version of this
content can go to www.bio.davidson.edu/icb where they will find
pedagogically-designed and interactive Integrating Concepts in Biology
for introductory biology college courses or a high school AP Biology
course.
Acknowledgments
Publishing this book would not have been possible without the gener-
ous gift of Dr. David Botstein who shared some of his Breakthrough
Prize with co-author AMC. Davids gift allowed us to hire talented artists
(Tom Webster and his staff at Lineworks, Inc.) and copyeditor Laura
Loveall. Thanks go to Kristen Mandava of Mandava Editorial Services for
project management and guidance. In particular, we are indebted to Katie
Noble and Melissa Hayban for their many hours and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to
administrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond,
Verna Case, and Barbara Lom who had confidence in us and encouraged
us to persist despite setbacks along the way.
Thanks to my wife Amy Brooks for her constant support during the
development of this textbook, and my daughter Evelyn for her endless
energy. Thanks to Malcolm Campbell for his steadfast resolve and opti-
mism. Without him, this book would not exist. Thanks to collaborator
Laurie Heyer for taking my sometimes half-baked math ideas and turn-
ing them into powerful and elegant Bio-Math Explorations. I learned a
lot from both of them. While the math is largely absent from this book,
our collaboration with her made this a better book. Nancy Stamp at
Binghamton University, and Bill Dunson and Richard Cyr at The Penn-
sylvania State University influenced me greatly in how I think as a scientist
and approach my teaching. Finally, I thank my students in Integrated
Concepts in Biology II, who enthusiastically participated in our experi-
ment to redesign introductory biology, starting with the text and ending
with a new approach to teaching biology.
Introduction
Individual cells are affected by a variety of factors and can disrupt the
function of other cells. Despite the focus on the cell as an individual
entity, no cell exists in isolation and many biologists study cells in the
context of the whole organism. In this book, the focus is on the popula-
tion, but cells are viewed as populations within multicellular organisms.
Over evolutionary time, some cell populations of certain species evolved
into colonies of interdependent individuals. From those colonies, mul-
ticellular organisms evolved. Within a multicellular organism, popula-
tions of similar cells exist and function as a unit. They make up tissues
and organs. Each cell type exists as a population of cells, and each has a
particular structure related to function. Some cells remain undifferenti-
ated, waiting for the signal to become active and develop into a particular
type of cell. These stem cells have some properties that are similar to
cancer cells. The description of these phenomena reflect the themes of the
concept of cells: All cells come from preexisting cells, cells maintain inter-
nal environments that differ from external environments, cell structure
defines cell function, and cells communicate with other cells.
CHAPTER 1
salivary gland
pharanx tongue
salivary gland
esophagus
liver
stomach
gall bladder
pancreas
duodenum
ileum
rectum
anus
the anus as solid waste, or feces. Accessory glands (also part of the diges-
tive system) secrete digestive juices into the canal through ducts. These
accessory glands consist of salivary glands, pancreas, liver, and its stor-
age organthe gallbladder. Data will be analyzed that were collected in
order to learn about the function of the specialized cells mentioned previ-
ously that aid in digestion and assimilation of nutrients.
Various organs within the digestive system function to assist with
digestion and assimilation. For instance, the esophagus is a tube that
primarily functions to transport food to the stomach, the stomach does
some digestion but is also a food storage organ, the small intestine is
where most assimilation occurs, and the large intestine is where the solid
waste is formed and stored until defecation. Some of these organs will
be examined at the cellular level to discover how populations of cells in
these organs function in the digestion and assimilation processes. The
digestive system of animals is quite complex with organs of the system all
interconnected and with the system connected to other systems, such as
4 CELLS IN TISSUES
the circulatory and nervous systems. In this chapter only a small portion
of the system will be examined in order to understand how cells function
within larger systems.
The process of digestion follows ingestion of food and begins with
the simple act of chewing. Food is then swallowed; it travels down the
esophagus and enters the stomach. For centuries, scientists struggled with
determining whether the stomach secreted an acid and if so what kind
of acid. In 1823, William Prout definitively answered this question for
several species of mammal. Prout quantified the acid by feeding rabbits,
horses, cows, and dogs and then immediately removing the contents of
their stomachs. (He had to kill the animals to do this.) He then repeatedly
exposed the contents to distilled water and combined the mixtures. Prout
divided the mixture into four equal portions. The first was evaporated
to dryness, burned at high temperature, and dissolved again in distilled
water. He used silver nitrate to react with the chloride salts in the solution
to determine the amount of chloride ion (Table 1). To the second portion
Prout used a known amount of potassium hydroxide to exactly neutralize
the solution, allowing him to determine the amount of free acid present.
In the third fraction, Prout added a large quantity of potassium hydroxide,
which will react and neutralize all hydrochloric acid (HCl) to potassium
chloride and water. He then determined total chloride in the same manner
as before, with silver nitrate. The final fraction was used to determine if
any other acids were found.
Prout did not find any other acid in stomach secretions. He was only
able to find HCl. Of course, as soon as it is known that the stomach pro-
duces HCl, scientists wondered where the HCl was made and how it was
produced given that HCl is so destructive and could potentially eat away
the stomach itself.
gastric pit
mucosa
gastric gland
submucosa parietal
cell
muscularis
externa chief cell
serosa
enteroendocrine cell
These gastric pits are indentations that lead to the gastric glands from
which the secretions come. The human stomach has several million of
these pits, and further into each pit scientists discovered other types of
cells, which have different functions; one of which will be examined.
Figure 2 shows parietal cells, located in pits in the upper part of the
stomach, which are the cells that produce HCl. Any macromolecule that
is affected by acidic conditions can begin to break down in the stomach
after HCl is secreted. The parietal cells thus have an important role in the
digestion of food.
When a person eats, their stomach becomes distended and proteins
begin to be broken down into component amino acids; substances are
released that activate parietal cells (Figure 3). These substances include
gastrin, histamine, and acetylcholine. Activation leads to a series of events
at the molecular level within parietal cells.
Muallem and colleagues performed a study to determine how stim-
ulation of the parietal cells affected parietal cell internal pH. If pH of
individual cells goes up, then it can be concluded that hydrogen ions are
being excreted from the cell and pH of the surrounding environment goes
down. The scientists used a centrifugation technique to isolate parietal
cells from the epithelium of the upper part of a rabbits stomach. Keep
in mind the structure of the parietal cell, especially the deep infolding on
the lumen end of the cell, which increases the surface area for secretion.
The measurement of internal cell pH in parietal cell suspensions was
achieved through use of a dye that can permeate cells. This dye can be used
neuron
A parietal cell
A
G cell
ECL
neuron
cell HCl
H
G G
circ
ula
tion
to estimate pH inside the cells. The scientists measured the emission inten-
sity of the dye at two different wavelengths; the ratio of the intensities is
pH-dependent. Knowledge of the intensity ratio in solutions of known pH
allows researchers to estimate intracellular pH. The scientists incubated cells
with dye for 20 minutes and then washed the cells so that any dye not taken
up by cells was washed away. Keep in mind that relatively small changes in
pH, which is measured on a logarithmic scale, in single cell suspensions
could actually lead to large decreases in pH in the stomach lumen.
Muallem and colleagues exposed cells to a medium that contained no
sodium ions (Na+) and found that intracellular pH went down (Table 2).
When they added Na+, the pH rapidly went up. Addition of a chemical
that inhibited the Na+/H+ exchange protein at the same time that Na+
was added completely inhibited the rise in pH, illustrating that Na+/H+
exchange is important for maintaining intracellular pH in the resting cell.
Next, Muallem and colleagues exposed cells to histamine, a known
activator of parietal cells (Table 2). The Na+/H+ exchange inhibitor was
added in one treatment before exposure to histamine. Histamine binds
histamine, and then the chemical that increases cAMP leads to a large
increase in intracellular pH. This suggests that the histamine-histamine
receptor interaction leads to an increase in cAMP levels in the cell.
The increase in cAMP levels within the parietal cell leads to other
changes. The entire scheme is complex and not of it all is shown here, but
ultimately acid secretion at the lumen end of the parietal cell occurs. The
Cl/HCO3 exchange of proteins at the end of the cell away from the
lumen is responsible for expelling HCO3 and bringing in Cl.
Recall that in the absence of Na+, intracellular pH dropped dramati-
cally because the parietal cell could not rid itself of excess H+. In the
presence of Na+, the resting, non-stimulated cell has an intracellular
pH of about 7.1. In the absence of Na+ but the presence of the H+/
K+-ATPase inhibitor and histamine, there is a slight rise in intracellular
pH. This rise is larger when there is just Na+ and histamine but only
after a lag period. The researchers concluded that the H+/K+-ATPase
was responsible for the further rise in pH to about 7.3. The action of
this pump expelling H+ into the lumen is what leads to production
of HCO3. Expelling HCO3 brings Cl into the parietal cell, which
then diffuses to the lumen end and is then secreted into the lumen. The
K+ also leaks back out through a separate channel protein after being
exchanged with H+.
The acid secretion at the lumen end of the parietal cell results
in higher activity of anion exchange, which is caused ultimately by
the higher intracellular pH due to higher HCO3. This leads to main-
tenance of cell homeostasis; researchers have found that after the initial
increase in intracellular pH, if the parietal cell continues to secrete H+
and Cl, intracellular pH does not continue to rise. The combined
function of both exchangers is necessary for optimal acid secretion.
The parietal cell clearly has specialized proteins to facilitate the pro-
duction of acid. It also has a specialized morphology (Figure 4). The rest-
ing state looks very different from the stimulated state.
In the resting state, deep infoldings on the lumen end of the parietal
cell are seen. The H+/K+-ATPase pumps are sequestered within vesicles
called tubulovesicles. Activation of acid secretion leads to two func-
tional changes: tubulovesicles fuse with the membrane of the infolded
region that opens to the stomach lumen, and this membrane acquires
10 CELLS IN TISSUES
tubulovesicles
at rest
mitochondria
infolding
stimulated
nucleus
infolding
lumen
muscle
layers lymphatic
villi vessel
GLUT-2
transport
protein
0 0
A sglt1+/+ sglt1/ B sglt1+/+ sglt1/
1200
600
1000
450 800
300 600
400
150
200
0 0
C glut2+/+ glut2/ D glut2+/+ glut2/
epithelial cell membranes (Figure 7). In the images, rows of epithelial cells
that line up to form the villi of the small intestine can be seen (see also
Figure 5). Note where SGLT1 localizes relative to the cell nuclei and how
that differs from the localization of GLUT2.
By knocking out the sglt1 gene in mice and then comparing
responses to wild-type mice, scientists can determine the effect of the
SGLT1 transmembrane protein. This is also true for the glut2 mutants.
14 CELLS IN TISSUES
A B
C D
Mutantsglt1/ mice take up very little glucose into epithelial cells and
show very little glucose in the blood relative to wild-type mice. The scien-
tists concluded that the function of SGLT1 was to transport glucose into
the epithelial cell. If glucose is not getting into the epithelial cell, then it
cannot find its way into the blood. The localization of SGLT1 near the
lumen further confirms this. The lack of SGLT1 in the mutants confirms
that the mutant really is deficient in producing that protein.
THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 15
Bibliography
Baron JH: The discovery of gastric acid, Gastroenterology 76:10561064,
1979.
Baumont W: Experiments and observations on the gastric juice and physiol-
ogy of digestion, Cambridge, MA, 1929, Harvard University Press.
16 CELLS IN TISSUES
Rder PV, Geillinger KE, Zietek TS, et al.: The role of SGLT1 and GLUT2
in intestinal glucose transport and sensing, PLoS One 9(2):e89977,
2014.
Kousoulis AA, Tsoucalas G, Armenis I, et al.: From the hungry acid to
pepsinogen: a journey through time in quest for the stomachs secre-
tion, Ann Gastroenterol 25(2):119122, 2012.
Langley JN, Edkins JS: Pepsinogen and pepsin, J Physiol 7(5-6):371415,
1886.
Prout W: On the nature of acid saline matters usually existing in the
stomachs of animals, Philos Trans R Soc Lond 1:4549, 1824.
CHAPTER 2
Populations of Endocrine
Cells in Animals Affect the
Whole Organism
are decapitated on the fifth day after blood feeding, according to Table 3,
but they do not all have the ability to induce molting in fourth stage
individuals decapitated after 24 hours. Most 24-hour decapitated indi-
viduals molted with the individual to which they were connected if that
individual was decapitated at or after the critical period. Wigglesworth
concluded that either there is a molting hormone secreted from the head
or nervous impulses that cause secretion by a gland posterior to the head.
Wigglesworth knew of several glands that were candidates for the
source of hormone. He took serial sections of the head and thorax of
fifth stage insects in all phases of the molting processfrom fasting to
10 days after feeding, and then from every other day until molting. Serial
POPULATIONS OF ENDOCRINE CELLS AFFECT ANIMALS 21
He found that both insects in the pair molted simultaneously and that
fourth stages molted into the fifth stage but with adult characteristics. He
then repeated the experiment using fifth stage individuals that had passed
the critical period connected to first stage individuals decapitated 24
hours after feeding. When these first stages molted into the second stage,
Wigglesworth found two-thirds of them to be diminutive adults.
In the preceding experiments (see Table 4) where fourth stage indi-
viduals decapitated after the critical period were connected to fifth stage
individuals decapitated 24 hours after feeding, the fifth stages that molted
became adults, but there were defects in the adult structures, most nota-
bly in genitalia that were imperfectly formed. The further past the critical
period the fourth stage individuals were, the stronger the effects were on
the development of adult structures as the fifth stage individuals molted
into adults. In a final set of experiments, fourth stage individuals that had
just reached the critical period were connected to each other, and it was
found that both individuals developed some adult characteristics when
they molted into the fifth stage.
When earlier stages past the critical period were connected to fifth
stage individuals decapitated 24 hours after feeding, the fifth stage indi-
viduals almost always molted, as seen in Table 4. This suggests that the
molting hormone is the same across all stages of development. Thus
there is a second hormone whose presence prevents metamorphosis in
early stages or whose absence causes metamorphosis in the fifth stage.
In fact, the hormone present in fourth stages past the critical period
caused defects in fifth stage individuals molting into adults, indicating
that it is the presence of the hormone, not its absence that prevents
molting from juvenile to adult. It did not prevent those fifth stage indi-
viduals from molting into adults, however, due to the difference in size
between fourth and fifth stages. Because fifth stage individuals are two
to three times larger than fourth stages, the cross-circulated blood is
diluted in the fifth stage. Thus, it is the absence of the hormone that
causes metamorphosis.
Wigglesworths next task was to determine the source of the hor-
mone, which is now called juvenile hormone. He performed decapitation/
connection experiments (Table 5) and examined serial sections of early stage
individuals as he did with fifth stage individuals. Recall that in fifth stage
POPULATIONS OF ENDOCRINE CELLS AFFECT ANIMALS 23
individuals during the critical period, cells in the corpus allatum become
swollen and stained pink. The scientist found that in earlier stages, a smaller
proportion of cells in the corpus allatum exhibited the change observed in
fifth stages. The remaining cells do not undergo this particular change.
The experiments described in Table 5 suggest that the head, specifi-
cally the corpus allatum, is the source of the hormone that regulates meta-
morphosis and that the hormone is different from molting hormone. If
just the tip of the head is cut off in a post-critical period individual, juve-
nile hormone is still produced, as indicated by the effects on individuals
decapitated prior to reaching the critical period. It appears that the head
is still necessary after the critical period in order to prevent metamorpho-
sis, and this is further supported by decapitation experiments on fourth
stages at compared to beyond the critical period.
The serial section results that differed between early stages and fifth
stages led Wigglesworth to conclude that the corpus allatum was not
only the source of molting hormone but was also the source of juvenile
hormone. Earlier it was stated that all cells swell and stain pink in the
fifth stage corpora allata. This is likely due to the high concentration of
molting hormone necessary to induce molting in a larger individual.
Because all cells swell, the scientist reasoned that all were producing
molting hormone; none were producing juvenile hormone. Wiggles-
worth further found that cells from the corpus allatum in early stage
24 CELLS IN TISSUES
individuals did not all stain pink, and he suggested that cells that do
not stain pink in early stages are the source of juvenile hormone. Later
experiments where he transplanted the corpus allatum from an individ-
ual past the critical period to a fifth stage that was decapitated 24 hours
after feeding confirmed this: Those fifth stages molted into another
juvenile stage, not the adult.
Although Wigglesworth hypothesized that the source of molting
hormone was the corpus allatum, he still need to perform experiments
to support his hypothesis. Several years after his initial experiments on
molting and metamorphosis, which were conducted in the early 1930s,
he performed more experiments on molting hormone. In these experi-
ments, Wigglesworth used more sophisticated dissection techniques than
the decapitation experiments conducted earlier. In these experiments,
he dissected out different parts of the brain, including different lobes,
nerves, and ganglia, masses of nerve tissue containing cell bodies of neu-
rons, from fourth or fifth stage individuals past the critical period and
transplanted those organs into the abdomens of fourth stage individuals
that were decapitated 24 hours after feeding (thus, they had not passed
the critical period; Table 6).
As stated earlier, individuals decapitated early do not molt unless they
are artificially provided with molting hormone. Wigglesworth reasoned
Bibliography
Wigglesworth VB: The physiology of ecdysis in Rhodnius prolixus
(Hemiptera). II. factors controlling moulting and metamorphosis, Q
J Microsc Sci s2-77:191222, 1934.
Wigglesworth VB: The function of the corpus allatum in the growth
and reproduction of Rhodnius prolixus (Hemiptera), Q J Microsc Sci
79:91121, 1936.
Wigglesworth VB: The determination of characters at metamorphosis in
Rhodnius prolixus (Hemiptera), J Exp Biol 17:201223, 1940.
Wigglesworth VB: The thoracic gland in Rhodnius prolixus (Hemiptera)
and its role in moulting, J Exp Biol 29:561570, 1952.
CHAPTER 3
other differentiated cells, including gut cells, skin cells, and neural cells.
Stem cells may go through particular developmental stages. They can
divide and produce more undifferentiated stem cells or they can divide
into tissue-specific stem cells. From there, they would divide into short-
term progenitor cells and then finally into the differentiated cells.
Populations of stem cells are self-renewing. Both the stem cells and
tissue-specific stem cells have the ability to produce more individuals of
that type. Self-renewal is a critical property of stem cells. If all stem cells
differentiated into other types of cells, there would be no stem cells left
to produce mature cells later in life. Once cells become progenitors, they
are incapable of self-renewal; they cannot produce more of themselves.
Second, stem cells have potential to become a wide variety of other cells.
That is, they are multipotent. However, once a stem cell develops into
a tissue-specific stem cell, the new tissue-specific stem cell cannot then
produce or revert to a multipotent stem cell.
How do stem cells know when to divide? A variety of substances regu-
late the cell cycle (Figure 9). The cell cycle has several phases; the phases
and sequence depicted in Figure 9 are the gap phase to DNA synthesis
phase, which then leads to mitosis. In the absence of appropriate signals,
cells may enter a quiescent stage where they are not in the cycle at all.
The proper signals bring the cell into the gap phase, and once it is past
the restriction point, the cell is committed to continuing the cycle. The
presence of growth factors, such as a transcription factor E2F, takes a cell
past the restriction point; otherwise development remains arrested. The
scheme in Figure 9 is highly simplified to illustrate the basic ways in which
the cell cycle is regulated. Proteins may either inhibit or stimulate other
proteins or their expression or the cell cycle. The dark grey oval protein
above the cell cycle bar inhibits the cell cycle unless it is phosphorylated
twice, represented by the small grey circles attached to the oval protein.
Of course, stem cells and tissue-specific stem cells have much more
complex interactions, so it does not represent all pathways in other types
of stem cells. There is a complexity and variety of stimulatory and inhibi-
tory effects within a typical stem cell.
A stimulatory effect could occur through increased expression of a
gene or by effecting a change in a protein, whereas inhibition could occur
through repression of transcription or binding to a protein. Stimulation
STEM CELLS AND CANCER CELLS 29
supression
protein
activation complex
and inhibition is often effected by two classes of genes and their proteins.
Proto-oncogenes code for proteins that regulate cell growth and dif-
ferentiation but can cause cancer when mutated or expression increases.
Tumor suppressors are genes or proteins that protect cells from cancer;
when mutated, they can cause cells to progress to cancer.
There are several pathways of interaction that control whether or not
DNA synthesis commences in stem cells. Once DNA synthesis commences,
the cell is on the path to mitosis, which in a stem cell leads to either dif-
ferentiation or self-renewal. The red symbol at the top of the cell represents
a chemical, often called a mitogen, a chemical substance that activates or
promotes cell division, bound to a membrane protein. That chemical is a
signal to the stem cell, and the binding of the chemical interactions among
proteins inside the cell. Mitogens stimulate both the cyclin D:cyclin-dependent
kinase 4 protein complex and phosphoinositide 3-kinase (PI3K).
In the absence of a mitogen, there are other gene products that con-
trol self-renewal of stem cells. Data on how the interactions in these
30 CELLS IN TISSUES
pathways have been determined in stem cells and whether they affect
self-renewal will be examined next. Components at or near the beginning
of the pathway are good places to start the investigation. Sean Morrison
and his colleagues studied the proto-oncogene Bmi-1. They bred colonies
of mice that were normal, heterozygous for normal and mutated Bmi-1,
and homozygous for the mutated Bmi-1. Morrison and his colleagues
hypothesized that loss of Bmi-1 would affect self-renewal of neural stem
cells. At three different stages of development, 14.5 day old embryos, at
birth, and 30 days after birth, the researchers sacrificed mice and exam-
ined specific areas of their brains (the central nervous system [CNS])
and tissues from the peripheral nervous system (PNS). The central ner-
vous system (CNS) is the brain and spinal cord that coordinates activity
of animals with bilateral symmetry, while the peripheral nervous system
(PNS) connects the limbs and organs to the CNS.
The scientists suspended cells from these nervous system tissues in a
liquid medium. They counted cells using a special cell sorter where cells
are labeled with fluorescent dyes. The dyes are attached to antibodies that
bind to particular cell types. The cell sorter detects and counts the cell
types. The cells were then plated into cell cultures at a low density so
that individual cells, if they grew into populations, formed distinct colo-
nies on the plates. They then counted the number of cells per colony to
determine proliferation and differentiation of individual cells. Regardless
of stage of development, they found similar trends in response, which
was that normal mice always had significantly more cells per colony than
Bmi-1 deficient mice.
Morrison and colleagues also determined the number of cells dying
in each culture and the percentage of 5-bromodeoxyuridine (BrdU) taken
up by each culture. BrdU is a synthetic DNA base that is an analogue of
thymidine and is used to detect cell proliferation. BrdU is incorporated
into newly synthesized DNA of replicating cells (the S phase), substituting
for thymidine during DNA replication. There were no differences in cell
mortality in comparisons of cells from the same type of mouse stem cell
(whether embryonic, newborn, or adult). However, BrdU incorporation
was always much higher in normal mice than in Bmi-1 deficient mice.
In culture, stem cells typically form spheres, making them easy to identify,
and the scientists measured the ability of these new cells to self-renew when
STEM CELLS AND CANCER CELLS 31
transferred to a new culture (Table 7). The numbers shown are the average
number of new stem cells generated from one transferred spherical stem cell.
The data show that stem cells self-renew and proliferate in the pres-
ence of functioning Bmi-1 gene. When the gene is not functioning the
gene product, Bmi-1, is absent from the stem cells; self-renewal, prolifera-
tion of colonies, and DNA synthesis are all drastically reduced in com-
parison to stem cells with Bmi-1. If stem cell self-renewal was not affected
but mortality was higher in the absence of Bmi-1, the scientists might
still have obtained the patterns they found. That is why it was important
to determine the percent mortality in the presence and absence of the
Bmi-1 protein. The results show that mortality was unaffected by the
loss of the gene and its protein, leading to the conclusion that it was self-
renewal and proliferation, not death of existing cells, that were altered.
It turns out that the protein Bmi-1 acts to suppress expression of the
genes p16Ink4a and p19Arf. Both of these proteins are tumor suppressor
proteins and, when functioning, normally prevent phosphorylation of
retinoblastoma (Rb). Note that Bmi-1 is not the only protein that sup-
presses expression of p16Ink4a and p19Arf. Hmga2 also suppresses them,
which helps explain why loss of Bmi-1 did not completely prevent self-
renewal. However, the absence of Bmi-1 should have predictable effects
32 CELLS IN TISSUES
recovery is partial because p19Arf is also involved in suppressing the cell cycle
through several intermediary proteins.
The components Bmi-1 and p16Ink4a are at or near the beginning of
the pathway. Studying the impact of their absence is a clever technique to
confirm their role in the cell cycle and stem cell self-renewal, and that is
exactly what Morrison and his colleagues did. There are other pathways
involved in regulating stem cell self-renewal, but this example points out
the complex controls on stem cell self-renewal.
Controls on self-renewal maintain populations of cells. Stem cells
have the ability for self-renewal, but self-renewal in many tissue-specific
stem cell populations is controlled by the cell cycle. Differentiation of
stem cells requires another set of complex controls. For any tissue that fre-
quently becomes damaged or is exposed to a high degree of wear and tear,
it would be expected to possess an active population of stem cells capable
of producing the cells that make up that tissue. Certainly blood, skin, and
the cells that line the alimentary canal would fit that category. Lung cells
are exposed to the external environment every time a person inhales, and
they are frequently exposed to pollutants in the air. Long-term exposure
to pollutants causes wear and tear on the tissues of the lungs.
Lung stem cell populations would be important in generating new
cells to replace damaged ones, and the controls on self-renewal and dif-
ferentiation should be similar to the controls in other stem cell popula-
tions. Lung cancer is a common and often fatal form of cancer. Is there
any relationship between the regulation of stem cell self-renewal and
cancer?
The genes and proteins involved in controlling the cell cycle were earlier
called proto-oncogenes and tumor suppressor proteins. With names like these,
there is likely a known relationship between these genes and proteins and the
formation of cancer. Daniel Betticher and a team of colleagues studied sev-
eral of the cell cycle genes and in human lung cancer. There are several dif-
ferent types of cancer that can develop in lungs. Betticher and his colleagues
studied non-small cell lung carcinoma (NSCLC; carcinoma is another term
for cancer), which makes up about 80% of all lung cancers. The name dif-
ferentiates it from small cell lung carcinoma (SCLC). Compared to SCLC,
NSCLCs are relatively insensitive to chemotherapy, which is why it is
important to understand how these cancers develop and grow. Their goal
was to examine lung cancers for expression of proteins involved in the cell
STEM CELLS AND CANCER CELLS 35
cycle, specifically some of the ones discussed earlier with regard to stem cell
self-renewal and the cell cycle: p16Ink4a, p19Arf, p53, and Rb protein.
The scientists obtained tumor samples from 49 patients who had
them removed surgically. The scientists obtained cells from each tumor
and created cultures of them in environmentally-controlled incubators.
In addition, the scientists cultured healthy lung tissue, a human leukemia
cell line (K562) known to have the p16/p19 gene locus (CDKN2) deleted
and two other cell lines known to contain and express CDKN2.
Cells from each culture were harvested and suspended in solution.
From the resulting suspension, they took a standard amount of pro-
tein. The scientists separated the proteins using gel electrophoresis. The
proteins were probed with antibodies specific to Rb, p16Ink4a, p53, and
p19Arf. In addition, the scientists also used antibodies to stain whole cells
from all cultures for these proteins. The scientists detected proteins using
a luminescence detection system; antibodies that bound to a particular
protein were luminescent on the gels and in the cells. Their controls were
analyzed to be sure that detection of proteins was accurate. Only three
of 49 tumors expressed normal levels of all four proteins; all others con-
tained abnormal expression (either downregulated, with expression lower
than normal, or overexpressed) of at least one of these proteins (Table 9).
At least one of the four cell cycle proteins was altered in 93.5% of the
tumors (see Table 9). Twenty-two had at least two of the genes, CDKN2
(p16Ink4a/p19Arf) and/or p53 altered, and for those 22, Rb expression was
normal. Normal lung tissue expressed both CDKN2 transcripts equally,
whereas only 57% of tumors expressed both transcripts equally (see
Table 11). That lower percentage was due to altered transcription of p16Ink4a.
Alterations in expression could occur from mutations in the gene or altered
signals that affect the expression of a gene that produces a particular pro-
tein product. The signal could turn on, off, up, or down the expression of a
gene. Recall in stem cells that loss of Bmi-1 reduced stem cell self-renewal,
and loss of p16Ink4a increased self-renewal. If p16Ink4a is downregulated or
mutated, it could cause unregulated growth. Cancer cells display unregu-
lated growth. Downregulation of both p16Ink4a and p19Arf should both
lead to relaxation of the cyclin complex suppression, which would lead to
phosphorylation of Rb. Once phosphorylated, Rb no longer suppresses
E2F, and this can take the cell into the synthesis phase of the cell cycle.
Altered expression of p53 and p16Ink4a both separately lead to effects
on Rb phosphorylation; the effect of alteration of both could be larger
than the effect of either alone, an example of an emergent property.
The two pathways involving p16Ink4a and p19Arf/p53 could cooperate in
tumor growth inhibition; and when either or both is altered, cells may
enter the cell cycle unchecked and become cancerous. Betticher and his
colleagues found that some lung cancer tumors with p16Ink4a alterations
also had p53 mutations. In lung cancers, both p16Ink4a and p19Arf are
frequently altered, and the scientists found a linkage between p19Arf and
p53. The negative association between p19Arf downregulation and abnor-
mal p53 expression suggests a common pathway in malignant growth and
38 CELLS IN TISSUES
supports the hypothesis that the p19Arf peptide plays a role in human can-
cer. The scientists concluded loss of two controls on the cell cycle, p16Ink4a
downregulation and aberrant p53 expression, leads to a high probability
of NSCLC tumor formation. Mutations in several of the genes involved
in the cell cycle are also known to play a role in a variety of other cancers.
Although some of the controls on the cell cycle in stem cells have been
discussed, it is important to determine whether these controls, or altera-
tions in them, play a role in developing cancer. In lung cancer, and pos-
sibly other cancers, alteration of the CDKN2 locus can simultaneously
impair both the p16Ink4a- cyclinD/CDK4-Rb and the p19Arf-p53-Cip/
Kip-cyclinE/CDK2-Rb pathways. When DNA mutates or cells fail to func-
tion normally for some other reason, tumors may begin to grow, depend-
ing upon other changes that have occurred. The connection between stem
cells and cancer cells is that the biochemical machinery that regulates self-
renewal and the cell cycle in stem cells contains many proteins that, when
altered, lead to unregulated growth. Cancer is a group of diseases char-
acterized by uncontrolled growth and spread of abnormal cells. If cancer
is considered as a disease of unregulated self-renewal, the connection to
regulation of stem cell self-renewal is clear. Tumors may originate from the
transformation of signaling pathways in normal stem cells.
Each cell type in a multicellular organism exists as a population of
cells, and each has a particular structure related to its function. Over
evolutionary time, populations of cells within multicellular animals have
evolved complex mechanisms of regulation, which can become dysfunc-
tional through mutation or improper signaling. Stem cells have no regular
day-to-day function and remain undifferentiated. They await signals to
proliferate and differentiate. Cancer cells have properties similar to stem
cells. In cancer cells, the proliferation signal may always be on; although
some types of cancers grow faster than others, the unregulated or dysfunc-
tional cell cycle is common.
are clearly requested by the patient, and may not directly affect health or
survival of the patient. However, if the surgery increases the individuals
self-esteem and attitude, then there may be an indirect benefit to their
health. More immediate emergencies may require rapid and extreme
interventions to keep a patient alive, such as when a patient has a heart
attack, stroke, or brain embolism.
Some interventions may be preventative, designed to stave off future
problems. Intervening after the fact may be easier for patients and physi-
cians. Consider an obese patient who has been told by his doctor to either
lose weight, or face heart disease and diabetes later in life. The patient
refuses to comply with a change in lifestyle, which would be a medical
intervention, but readily accepts insulin and heart medication after he
has developed diabetes and heart disease. So some types of interventions
may be more readily accepted, such as when there is a life-threatening
emergency or when a prescription drug can be used in place of a change
in lifestyle.
Other interventions (such as, organ transplants) are not so easy, and
healthy organs for transplant may be in short supply. For organ trans-
plants there are specific principles that the medical community adheres
to in order to most ethically share the scarce resource of healthy organs.
Principles used to decide on the use of these sorts of medical interven-
tions can be categorized according to several fundamental values: treat-
ing people equally, favoring the worst-off, maximizing total benefits, and
promoting and rewarding social usefulness.
Equal treatment may be accomplished through a lottery or through a
first-come, first-served principle. The worst-off patients may be the sickest
or the youngest. Those who are sickest are currently suffering; those who
are youngest may be prioritized because they have the most years in front
of them. When maximizing total benefit, principles may be used to save
the most lives or those patients with the best prognosis for benefitting
from the intervention. Finally, promoting and rewarding social usefulness
provides priority to those who are in the best position to help society in
the future or have greatly helped society in the past and have suffered for it.
According to some ethicists, some of these principles are flawed, in
that they recognize ethically irrelevant considerations. Other principles
STEM CELLS AND CANCER CELLS 41
Bibliography
He S, Nakada D, Morrison SJ: Mechanisms of stem cell self-renewal,
Annu Rev Cell Dev Biol 25:377406, 2009.
Hook CC, Mueller PS: The Terri Schiavo saga: the making of a tragedy
and lessons learned, Mayo Clin Proc 80(11):14491460, 2005.
Persad G, Wertheimer A, Emanuel EJ: Principles for allocation of scarce
medical interventions, Lancet 373(9661):423431, 2009.
Reya T, Morrison SJ, Clarke MF, et al: Stem cells, cancer, and cancer stem
cells, Nature 414(6859):105111, 2001.
42 CELLS IN TISSUES
gastric pits. Indentations in the stomach that mark entrances to the gastric glands.
hemolymph. Hemolymph is the body fluid of insects.
histamine. A chemical released by cells in response to injury and in allergic and
inflammatory reactions, causing contraction of smooth muscle, dilation of capil-
laries, and known to activate parietal cells in the stomach.
homeostasis. maintain internal conditions within a range of acceptable extremes.
hormones. Hormones are chemical substances, usually peptides or steroids,
produced by one cell type, that control and regulate activity of other cells.
information. Information is stored, communicated, and implemented or
interpreted.
ingestion. Ingestion is the process of taking food into the body.
juvenile hormone. An insect hormone that regulates larval development and
inhibits metamorphosis.
large intestine. An organ of the digestive system of mammals containing the
cecum, colon, and rectum.
liver. An organ of the digestive system responsible for detoxification, carbohy-
drate metabolism, and other metabolic processes.
lumen. Lumen is the inner open space or cavity of a tubular organ.
lymphatic vessels. Lymphatic vessels are thin walled, valved structures that carry
lymph, a colorless fluid that contains white blood cells.
metamorphosis. Metamorphosis is an abrupt developmental change in the form
or structure of an animal.
mitogen. A mitogen is a chemical substance that activates or promotes cell
division.
molting. The process of shedding a cuticle in insects and other invertebrates with
exoskeletons during development.
mucus. A slimy substance, typically not miscible with water, secreted by mucous
membranes and glands for lubrication and protection.
multicellular. Organism consists of many interdependent cells, which cannot
exist on their own.
multipotent. Multipotent cells have potential to differentiate into any number
of other cells.
muscle cells. An elongated contractile cell that forms muscles.
neurons. The basic cell type of the nervous system of animals that transmits nerve
impulses.
neurosecretory cells. Neurons that function to translate neural signals into chem-
ical signals by secreting neurohormones.
organisms. An individual animal, plant, or single-celled life form.
pancreas. the organ in your abdomen that makes insulin, among other compounds.
parietal cells. Specialized epithelial cells in the gastric glands of mammalian
stomachs that secrete hydrochloric acid.
pepsin. An enzyme that breaks down proteins.
GLOSSARY
47
pepsinogen. A protein secreted by the stomach and converted into the enzyme
pepsin.
peripheral nervous system. The peripheral nervous system (PNS) connects the
limbs and organs to the CNS.
peripheral nervous system (PNS). The peripheral nervous system (PNS) con-
nects the limbs and organs to the CNS.
peristalsis. Muscular contractions that move food through the alimentary canal.
population. A population is a group of individuals of the same species living in
the same place at the same time
prothoracic gland. An endocrine gland in insects and related invertebrates that
secretes molting hormone.
protocerebrum. The first segment of the brain in an insect that innervates the
compound eyes.
proton pump inhibitor (PPI). A chemical that inhibits the activity of proton
pumps.
proto-oncogenes. Genes that code for proteins that regulate cell growth and dif-
ferentiation but can cause cancer when mutated or expression increases.
salivary glands. Glands in the digestive system of animals that secrete saliva.
second messenger. An intracellular chemical that mediates cell activity by relay-
ing a signal from an extracellular molecule bound to the cells surface.
small intestine. An organ of the digestive system of mammals responsible for
much digestion and assimilation of nutrients.
smooth muscle. Smooth muscle is a contractile type of muscle tissue controlled
by the involuntary nervous system, occurring in the walls of the stomach, intes-
tines, and blood vessels.
stem cells. Long lived, produce daughter cells. and replenish themselves indefinitely.
stomach. An organ of the digestive system responsible for some digestion and
temporary storage of food.
tissues. Collections of more than one cell type of a single species that work
together to perform a complex function.
tubulovesicles. Tube-shaped vesicles in parietal cells.
tumor suppressors. Tumor suppressors are genes or proteins that protect cells
from cancer; when mutated, they can cause cells to progress to cancer.
tumors. Abnormal growths of tissue.
undifferentiated. Not differentiated into a particular cell type, as in an undif-
ferentiated stem cell.
Index
Accessory glands, 3 Fats. See Lipids
Alimentary canal, 1, 2 Feces, 15
mammalian digestive 5-bromodeoxyuridine (BrdU), 30
system, 23
Antagonist, 8 Gallbladder, 3
Assimilation, 1, 2 Ganglia, 24
Gastric pits, 56
Betticher, Daniel, 34, 37 Gastrin, 8
Bicarbonate, 10 Glucose absorption and transport, 12
Bmi-1, 3032 Glucose transporter 2 (GLUT2),
effects on neural stem cells, 3334 1215
loss effect on self-renewal of CNS/ Gut. See Alimentary canal
PNS stem cells, 31
Hematopoietic stem cells (HSCs), 27
Cancer cells, 27, 38 Hemolymph, 19
ethical, legal, social implications of, Histamine, 78
3841 Homeostasis, 2
versus stem cells, 2741 Hormones, 17
Capillaries, 12 juvenile, 22, 23
Carbohydrates, 12 molting, 20, 22, 24, 2526
Carcinoma, 34 Human digestive system, anatomy
CDKN2, 32, 33, 35, 36, 38 of, 3
Central nervous system (CNS), 30, 31
Connective tissue, 5 Information, 1
Corpus allatum, 21, 23 Ingestion, 1, 2
Critical period, 19
Cyclic adenosine monophosphate Juvenile hormone, 2224
(cAMP), 89
Large intestine, 2
da Vinci, Leonardo, 2 Lipids, 11
Differentiated cells, 27 Liver, 3
Digestion, 1, 2 Lumen, 5
Luminescence detection system, 35
Endocrine system, 17, 25 Lung cancer, 34, 35, 36, 37
Epidermis, 18 Lymphatic vessels, 12
Epithelial cells, 5
small intestine, 11, 15 Medical intervention decision-
Epithelium, 5 making, ethical, legal, social
Esophagus, 2 implications of, 3841
Exoskeleton, 17 Metamorphosis, 21
molting of, 17, 18, 20 Microvilli, 12
50 INDEX
The Momentum Press digital library is very affordable, with no obligation to buy in future
years.
For more information, please visit www.momentumpress.net/library or to set up a trial in the
US, please contact mpsales@globalepress.com.
EBOOKS Cells in Tissues
FOR THE Christopher J. Paradise A. Malcolm Campbell
APPLIED BIOLOGY COLLECTION
Two systems illustrate how individual cells of an organ system
SCIENCES
function, communicate, and coordinate activities. The digestive
LIBRARY system breaks down and absorbs nutrients, and some special-
Create your own ized cells break down and absorb nutrients. The case of parietal
Customized Content cells in the stomach and epithelial cells in the small intestine
Cells in Tissues
Bundlethe more are used to describe how cells function as a unit within organ
books you buy, systems, coordinating activities and communicating with one
the greater your another. The endocrine system of insects affects molting and
discount! metamorphosis, and specialized cells are also important in each
of these processes within that organ system. The experiments
that were devised to determine the role of hormones in insect
THE CONTENT
molting and metamorphosis are described. Finally, stem cells
Energy Physics are healthy components of several different systems in animal
Engineering bodies and are described in relation to a disruption in function.
Biotechnology In this breakdown of function, cancer cells, in contrast to stem
Biology cells, can abnormally affect cell cycle regulation.
Mathematics
Christopher J. Paradiseis professor of biology and environ-
Chemistry
mental studies at Davidson College. He teaches introductory
biology, ecology, entomology, and topical seminars on ecotoxi-
THE TERMS cology and renewable natural resources. He also occasionally
Perpetual access leads a study abroad program in India. His research evaluates
for a one time fee anthropogenic factors that influence insect biodiversity at a
No subscriptions or variety of scales. His current research interests include effects of
access fees land use patterns on pollinator communities in parks.
Unlimited
A. Malcolm Campbellteaches biology at Davidson College,
concurrent usage
NC. He received national and international education awards:
Downloadable PDFs Genetics Society of America (2013); American Association for the
Free MARC records Advancement of Science (2012); and American Society for Cell
Biology (2006). He was the founding co-editor in chief of CBE Life
For further information,
Sciences Education; founding director of Genome Consortium
a free trial, or to order,
contact: for Active Teaching (GCAT); and member of the American Soci- Christopher J. Paradise
sales@momentumpress.net ety for Cell Biology governing council (20122014).
A. Malcolm Campbell