The Integument and

Connective Tissues

Week 3

An introduction to microscopic anatomy

The objectives of the lab are:

To introduce you to basic aspects of

histology, the study of tissues.

To study the integument at the gross and

microscopic levels.

To examine bone and cartilage and gain

an understanding of the major
characteristics of these tissues.

To introduce you to the morphology of a

vertebra (as opposed to vertebrate)

Diversity:
There is no diversity exercise, but instead you will
have a chance to look at models of developmental
stages of several different vertebrates.

Week 3

INTRODUCTION
In todays lab well do a number of different activities. You will see microscopes set up at each
station. This should be your first activity to go through the microanatomy of the integument and
then of bone and cartilage. Please read the appendix, which has a brief introduction to using the
microscope. Also take time to set yourself up. Adjust your chair so you can comfortably see
through the microscope and adjust the width between the eyes so that you can see through both
oculars. You will see much more, and be more comfortable, if you take the time to do this.
Remember histology is the study of tissues, the anatomical level between cells and organs. Tissues
provide the building blocks of the body. This will be the only lab in which histological study is
central, but subsequent labs will have optional histological exercises. If you continue into any
medical field you will spend considerable time understanding histology as it is important in
pathology and clinical diagnosis. Please make sure you read the targeted reviews on Bone
and Cartilage as well as on Histology for overviews of this material.

Par t I: The Integument

Recall from lecture that the integument or skin of vertebrates has
a dual origin embryonically: the epidermis develops from
epidermal ectoderm, the dermis from mesoderm. All vertebrates
possess both an epidermis and a dermis. The dermis is lacking
in cephalochordates and urochordates as well as other
invertebrates.
The epidermis, as a tissue that is a covering sheet, is an example
of an epithelial tissue. It may be simple (one cell layer) but is
generally squamous (many cells thick). The integument also
frequently contains glands which are also ectodermal in origin,
even though they may lie in the dermal layer. Keratin, an
extracellular matrix protein, is a product of the epidermis.
Compare sections of the skins of frogs (as representatives of
semiaquatic animals) and terrestrial mammals. The dermis does is
always composed of a variety of loose and dense connective
tissues. However, in different sections of different animals and
parts of the body you will see differences in the thickness of the
keratinzed layer, as well as the epidermis and dermis.

Remember, there are 4 major types

of tissue
1. Epithelial tissue (epithelium) is
specialized for protection of surfaces,
absorption, and secretion.
2. Connective tissue is specialized
for joining other tissues together and
it wraps and supports individual
organs and the body as a whole.
Connective tissue is characterized by
large amounts of extracellular matrix.
3. Muscular tissue is specialized for
contractility and conductivity. Well
talk about the different types later in
the course.
4. Nervous tissue is specialized for
irritability and conductivity. In
general, nervous tissue is highly
receptive to stimuli, and when
irritated, conducts waves of excitation
over long distances.

Look first at slides of frog skin (see illustration on next page).

The frog epidermis is thin and non-keratinized. Large glands,
Many sources define blood tissue
derived from the epidermis, project into the dermis. In favorable
(largely blood cells) as a 5th tissue
sections these will be seen to communicate with the surface via
type.
ducts. Most glands secrete mucous, which is important in keeping
the frogs skin moist. In some species toxic substances are also
secreted by specialized glands. Large, dark-staining cells between the epidermis and the glands
are chromatophores: cells containing pigment which give the animal color. Many fish,
amphibians, and reptiles have the ability to change color by expanding or contracting the
pigment granules in the chromatophores. The dermis in frogs is fairly unspecialized.

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Bio 330 L Fall, 2014

Note: there are two

different frog skin
specimens, one is stained
pinkish (as shown on the
top). The other is stained
with a different stain and
looks blackish brown (the
difference is due to
preparation and not species
differences). Make sure you
look at both of them, and
identify similar structures in
each. The specimens stained
blackish are of a species
that has many more mucous
glands. The specimen has also been prepared
such that a lot of the dermis has been removed.
Look at both and compare.
Label the glands, pigment cells, epidermis and
dermis in the slide to the left.

Next well look at several specimens of mammalian integument. The mammalian epidermis
(and that of most amniotes) is much thicker and is generally covered with a layer of keratin. Its
thickness, as well as the thickness of the keratin layer correlates primarily with the degree of
abrasion that part of the skin receives.
In general, the dermis is also much thicker
in mammals. The specific structure of the
dermis depends on the function of that
part of the integument, as we discussed
in class. Look at the following slides.
Weve labeled some structure for you
make sure you identify and label the
following on each: epidermis, dermis,
keratin. Label any glands, hairs or
pigment cells you observe as well.
1) Human abdominal skin.
Identify keratin, epidermis and dermis in
the human abdominal skin. Note the
thickness of the dermal layer.

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Week 3

2) Monkey skin
Examine the section of the
monkey skin. Notice the hair
cells, sebaceous glands and
adipose tissue. Adipose tissue
was also present under the
human abdominal integument,
it was removed before
sectioning.

3) Integument of the foot

Compare the skin from the sole of the foot in a human and cat to the abdominal skin. Note the
thickness of the epidermal layers and in particular the thick keratin layer. When you develop a
callus, you are thickening this layer. Label the two sections below. The difference in color is due
to staining technique.
Human foot

Cat foot:

The staining in the cat renders the dead, keratinized epidermal cells bright yellow-green. Below
this the brown layer consists of the germinative epidermal cells. The blue-green layer under the
epidermis is the dermis. There are also copious fat deposits in the pads/soles of the foot.
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Bio 330 L Fall, 2014

Now look at other structures such as bird feather (see illustration on the title page of a bird
feather) and the human fingernail.
Now, take a break from the microscope and explore the various structures of the integument laid
out for your inspection. Look at the quills of the African (big) and North American (smaller)
porcupine. What structure are these homologous with? Note the armadillo armor. What layer is
this derived from? Look at the baleen. Below weve shown a picture of how it fits in the whales
mouth. Watch the video of the week on whale feeding. Examine other structures on display.
Photos of fingernail slide. Use key below to understand the anatomy. (Left fingernail tip; right
fingernail base).

Whale baleen, used in filter

feeding whales.

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Par t II Connective tissues car tilage and bone

Connective tissues are classified primarily on the basis of the relative amounts of the different
structural components that are present. These components include cells of various types and also
extracellular components.
A distinctive feature of connective tissue is the predominance of extracellular materials.
There are two major categories of extra-cellular components: ground substance and fibers (which
are produced by specialized cells called fibroblasts). Fibers include collagen fibers, which do not
stretch and elastic fibers, composed of the protein elastin, which can stretch to 1.5 times resting
length, as well as other fiber types.
Cartilage is a specialized form of
connective tissue which has a firm and
definite shape essential to its
supporting function. It consists of a firm
matrix of collagenous fibers
surrounded by an amorphous matrix
consisting of a protein-polysaccharide
complex (proteoglycans).
Chondrocytes (=cartilage cells) lie in
lacunae (small cavities in the
extracellular matrix). Cartilage is a
major tissue in the skeletal system.
There are three major types of
cartilage, but today you will primarily
look at hyaline cartilage, which is
present in developing bone and also on many joint surfaces.
Bone is another specialized form of connective tissue and is the tissue which forms most of the
adult skeletal system in the majority of vertebrates. The matrix of bone is very solid due to the
deposition of calcium phosphate and other salts on the collagenous fibers. Bone tissue may have
either of two textures: Spongy or cancellous bone consists of irregular trabeculae (beams) of bony
tissue between which lie connective tissues and blood vessels. This is the first type of bone which
forms during the development of dermal and cartilage replacement bones. Spongy bone is often
reworked to form compact or lamellar bone. Compact bone consists of dense layers with few
interspaces. Bone tissue of both of these textures may be formed either as endochondral
ossification of as membrane bone.
Dermal or membrane bone has an intramembranous origin, i.e. its development begins with the
secretion of collagen by strands of mesenchymal cells in connective tissue membranes. Osteoblasts
(bone forming cells) differentiate and are responsible both for secreting the organic intercellular
substance and directing the deposition of calcium salts into trabeculae.
Cartilage replacement or endochondral bone is structurally similar to dermal bone, but is formed
by replacement of a cartilaginous model. The development of a skeletal element such as the
humerus begins with the formation from mesenchyme of a cartilage precursor. Later in
development the cartilage is invaded by periosteal tissue derived from the initial perichondrium.
The cartilage model is eroded away and replaced by a bony element. During development the
surviving fragments of cartilage serve as a framework for the deposition of bone tissue.
Make sure you review the glossary in the targeted review on bone and cartilage.
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We will look at a number of examples of bone and cartilage so that you can see the major
differences between these tissues.
Look first at Slide I-50 which is an example
of spongy bone, forming in a membrane.
The calcium phosphate has been removed
chemically leaving behind the collagenous
fibers (stained red) and osteocytes (bone
cells) trapped in lacunae. The bony tissue is
seen as an open meshwork of trabeculae.
In these sections you can see the bone tissue
as the dark reddish network. This is
probably the development of a bone of the
cranial roof (in a rodent); you can see a
suture between two bones forming. Look at
the illustration on the next page and make
sure you identify the trabeculae of
membrane bone and the osteocytes, which
are responsible for secreting the matrix.

On slide I-49 or I-44, the formation of cartilage replacement bone can be seen. This is a section
of a developing limb of a rodent, so you can see cartilage, joints and endochondral bone
formation. Youll also be able to see some elements of the integument. Take some time and study
both the cartilage as well as the bone. Bone formation is taking place in the center of the
elements. Notice the difference in texture of the newly formed bone: it is spongy centrally and
compact around the shaft of the bone, or bone collar (this is called cortical or perichondral bone).
Also, observe the changes in the character of the cartilage as it is being eroded away by the
process of ossification.
See the illustration on the below for an overview and photos of the section on the next page.

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Week 3

On slide I-45 you can see Haversian systems in mature, compact bone. The bone matrix is intact
and the section was made by grinding. This section is of compact bone, which has been
remodeled. Osteocytes (bone cells) lie in the lacunae and connect via small radiating canaliculi
(= little canals). Frequently it will be seen that groups of lacunae are concentrically arranged
around open central canals (Haversian canals). This arrangement reflects the concentric
deposition of matrix in layers or lamellae around a blood vessel. Review the structure of
Haversian canals in your textbook for more information.

Refer to the targeted review on bone and cartilage for more detail.

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Bio 330 L Fall, 2014

Par t III The anatomy of a ver tebra

In preparation for lecture on Thursday
we would like you to gain a basic
familiarity with a typical vertebral
body. Well see in lecture and in lab
next week that vertebrae vary along
the spinal cord and also are very
different in different species, but for
now wed like you to become familiar
with the terminology below.
The illustrations are of human thoracic
vertebrae, youll be given a cat thoracic
vertebra. Use your dissection guide for
details on cat anatomy. It is difficult to
see some features on two dimensional
illustrations, so look at the vertebra
from several angles. Please make sure
you know what we refer to when we
talk about:

Vertebral body or centrum

Neural arch,
Spinous process
Transverse process
Vertebral (neural) foramen
Superior articular process (and facet) =
prezygapophysis
Inferior articular process (and facet) =
postzygapophysis
Intervertebral foramen

After looking at a single vertebra, fit several

together and look at the way that the superior
and inferior processes (the terminology used in
human anatomy, the comparative terms are preand postzygapophyses) fit together to provide
torsional stability.
Illustrations from the following website:
http://biology.clc.uc.edu/fankhauser/Labs/Anat
omy_&_Physiology/A&P201/Skeletal/selected_
bones/Bone_Features.html

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W
Week 3

Once youu finish exam

mining an individual cat vertebra
v
and
d can identifyy all of the a
above elemeents,
look at thhe articulated skeleton. We will be coming
c
backk to this next week (and d
discussing it oon
Thursday), but notice that in a ma
ammal (and in
i most amniootes) we seee 5 distinct reegions:
Cervical vertebrae co
onnect the he
ead to the body they l ack large tra
ansverse proocesses, but ooften
have well developed spinous processes.
Thoracic vertebrae possess ribs; most
m connect to the ventrral sternum
Lumbar vertebrae
v
lacck ribs
Sacral ve
ertebrae havve specialize
ed form and facets to artticulate with the pelvis
Caudal vertebrae
v
are varied in number
n
and size in verteebrates theey make up the tail.
We will discuss
d
all of these in gre
eater detail next
n
week, foor now, simp
ply note the d
differences.

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Bio 330 L Fall, 2014

Par t IV Diversity of ver tebrate embryos

As you recall from lecture the life of any individual vertebrate begins with the union of a male
and a female gamete; it moves along a continuum of processes that produce an adult and
ultimately ends in death. It is important to remember development is a continuum, but it is useful
for us to recognize defined stages, like the frames of a motion picture film. As we discussed in
lecture, the major developmental stages of the vertebrates are these:
A. Embryonic stages
(1) Zygote: the one cell stage resulting from sperm fertilization of the egg (Gr. zygotes = yoked
together, i.e. union of egg and sperm).
(2) Early cleavage: 2 cell, 4 cell, 8 cell, etc. stages; cleavage partitions the egg into many smaller
undifferentiated cells called blastomeres.
(3) Blastula: a stage in which the blastomeres generally become rearranged around a fluid-filled
cavity. (L. diminutive of Gr. blastos = bud)
(4) Gastrula: the first major rearrangement of cells occurs during gastrulation (Gr. gastrula =
little stomach). This process establishes in the embryo the "tube-within-a-tube" configuration
around which the mature body is fashioned. The gastrulation process also establishes the three
germ layers from which the primordia for all organs and tissues of the adult body will form.
(5) Neurula: in this stage dorsal folds of ectoderm elevate and unite to form the neural tube. The
process is called neurulation.
(6) Pharyngula: all or most of the organ systems are established as rudiments, especially visceral
arches, in their proper positions at this stage.
(7) Fetal stages (largely in organisms in which no larval stage exists) in which all systems are laid
down, but the embryo continues to grow and further differentiate.
At the diversity table we have placed models of these stages from several different types of
organisms. There are models of the early stages in amphioxus, representing the primitive
condition. Models in frogs will demonstrate cleavage through neurulation in species with large
amounts of yolk. Then, we have models of chicken embryos in later stages, so you can see the
elements present in a pharyngula stage embryo. Finally, look at the embryos of pigs, which show
you a number of fetal stages in a mammalian embryo. Use these to review these stages and
understand some of the diversity in early development we see in vertebrate embryos.
For more information
Be sure to look at targeted reviews on Histology and also on Bone and Cartilage. Look at the
clinical correlations on vitamin D and also on Osteoporosis. Review the anatomy shown in the
histology atlas.
You can further review histology on the following website.
http://www.kumc.edu/instruction/medicine/anatomy/histoweb/index.htm
Remarkable videos of humpback whales feeding. This is notable not only because it shows you
how the baleen is used, but also the adaptations for expansion of the throat. Read the caption
below the video.
http://www.arkive.org/humpback-whale/megaptera-novaeangliae/video-08b.html
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Key terms:
You should understand the following terms. By understand we mean you should both be able to
define and also find the structure (where applicable) on a slide or specimen. Refer to the
targeted review for more detail on bone and cartilage.
keratin, epidermis, dermis, bone, cartilage, endochondral bone, membrane bone, Haversian canal,
osteocyte, chondrocyte.
vertebral body or centrum, neural arch, spinous process, transverse process, vertebral (neural)
foramen, superior articular process (and facet) = prezygapophysis, inferior articular process (and
facet) = postzygapophysis, intervertebral foramen
You should also be able to identify all the key structures we point out (such as keratin, glands,
chromatophores, fat pads, etc) in the histological slides you have examined.

Key questions:
What kinds of adaptations do we see in the integument of animals for protection against
abrasion or water loss?
Why might an animal not want to have an epidermis that has a thick keratin layer?
What characterizes connective tissue and what are the different kinds of connective tissues?
What are the key differences between cartilage and bone structurally and functionally?
What are the origins of cartilage and bone in evolution and in development?

Key competencies:
You should be able to identify major structures of the epithelium of a frog and mammal on a
histological slide (or photo of one).
You should understand what elements are derived from dermis and what elements from epidermis.
You should be able to identify cartilage and bone as well as the elements of the Haversian system
in a section (or photo of one).
You should know the difference between endochondral and dermal (= membrane) bone.
You should know the major anatomical elements of a typical mammalian vertebra.

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APPENDIX: GUIDELINES FOR USING A COMPOUND MICROSCOPE

We ask that when you use a compound microscope you follow these guidelines carefully. This is
important in order to optimize the performance of the microscope (so you can actually see
images), and to protect both the slides and the microscope itself. Many of the slides youll look at
are quite rare or expensive to replace.
Look at the illustration of the microscope below
(yours will be slightly different). The focusing knobs
(FK) have both a fine and a course focus. You can
use the ring controlling the diaphragm at the base of
the condenser to provide a larger circle of light if
necessary (CR).
Look at the array of objectives on your microscope
the power of the objective is identified on each
objective. For most specimens, you should use 10X
power, although you may want to start out with X4 to
find the specimen. When moving from one objective
to another, never use the objectives to rotate the
array always move it by grasping the objective
ring (OR) and gently rotating.

OR

FK

CR
Before using the microscope make sure you adjust its
position. Make sure your chair is at a proper height
to comfortably look through the eyepieces. Change
the distance between the two eye pieces so that you
can look through both comfortably. You may have to
adjust the eyepiece focus. Note that one eye piece, generally the one on the left, has a focusing
ring. Bring the specimen into focus while looking only through your right eye using the focusing
knob. Then open your left eye and use the eye piece focusing ring to bring the left eye into focus.
When changing slides, always move the stage DOWN (using the focusing knobs) and make sure
you are using an objective of 10X or less. After putting the slide on the stage, move the stage
back into focus. For some of the slides it may be useful to use a low power (4X) to scan the slide
for the section, and then move into higher powers for observation.
Always move the stage slightly DOWN before switching to an objective of HIGHER power.
Never touch a lens surface with anything (finger, tissue paper, etc). If it is dirty, call an instructor.
Note: the dissecting microscopes, which are used for observing whole mounts, are much more
forgiving of abuse, however, still be careful, and in particular avoid touching lens surfaces.

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