LAB EXERCISE 1 - Spermatogenesis

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ABI 3110 – Developmental Biology Laboratory

Laboratory Exercise No. 1
GAMETOGENESIS
The reproductive cells, called gametes, develop in the gonads. More specifically, spermatozoa
(or sperm) form in the seminiferous ampullae or seminiferous tubules of the testis, and ova (or eggs)
form in the follicles of the ovary. The process of sperm formation is called spermatogenesis, the process
of egg formation is called oogenesis, and together these events are termed gametogenesis. Two major
processes oc-cur during gametogenesis: (1) reduction division, or meiosis, and (2) the acquisition of
cellular specializations needed for fertilization and embryogenesis.
During gametogenesis, the gamete-producing cells undergo two meiotic divisions, so that
mature gametes are haploid; that is, each has only a single set of chromosomes rather than the double
set present in other body cells. These two meiotic divisions are called meiosis I and meiosis II. Terms
used to describe gametes during their differentiation are based on their meiotic state. Precursor cells
of spermatozoa are spermatogonia; those for eggs are oogonia. When these cells enter into meiosis I,
they are termed primary spermatocytes and primary oocytes, respectively. In males, completion of
meiosis I yields two secondary spermatocytes; completion of meiosis II yields four spermatids.
Spermatids are haploid (1N), which means that they have half the diploid complement of
chromosomes. The spermatids continue to differentiate during the process known as spermiogenesis
into four mature spermatozoa. In females, completion of meiosis I yields one secondary oocyte and the
tiny first polar body; completion of meiosis II yields a single haploid ovum and the second polar body
(which is also haploid). The first polar body may also undergo meiosis II to yield two tertiary polar bodies
that are genetically equivalent to the mature ovum, although they lack any reproductive future because
their tiny size limits the amount of yolk and other materials needed for normal development.
Gametes are highly specialized cells, the structures of which closely match their functions.
Spermatozoa are specialized for mobility, whereas ova are essentially packages of materials and
information that will be needed by the developing embryo. Thus, there is great disparity in the sizes of
sperm and egg cells and a corresponding difference in the metabolic cost of producing them. Because
of their smaller size, spermatozoa can be produced in extremely large numbers, and it is often the
number of eggs that limits fecundity in species of vertebrates.
SPERMATOGENESIS
Spermatogenesis is the development of spermatogonial cells to mature spermatozoa. The
process takes place within the seminiferous tubules boundaries of the testis. This process involves
mitotic cells division followed by meiosis which happens among spermatogenic cells. In mammalian
spermatogenesis is divided into three phases: mitosis, meiosis and spermiogenesis. The first
proliferative phase, the spermatogonia undergo series of mitotic divisions to ensure self-renewing
spermatogonial cells. In the meiotic phase, spermatocytes undergo two consecutive cell divisions to
produce haploid spermatids. In the third process, the spermatids undergo morphological changes to
develop into spermatozoa.
A unique characteristic of mammalian spermatogenesis is the developing germ cells form
stages, constituting the seminiferous epithelium cycle. The cycle refers to the different cellular
associations that appear in a specific section of the tubule at a given period of time. Each cellular
association is made of cells at different stages of spermatogenesis. All the stages of one cycle can also
be seen taking place in sequence within a long segment of the tubule which is now refer to the wave
of the seminiferous tubule (Herrera et al. 2003). In rats, the seminiferous epithelium cycle is divided
into 14 stages (I to XIV), with spermiogenesis, defined as the morphological transformation of
spermatids into spermatozoa, broken down further into 19 differentiation steps (from 1 to 19). A wave
encompasses all 14 segments in rat, 12 in mouse, and 6 in human consisting of various cell associations.
A segment is defined as a longitudinal portion of seminiferous tubule corresponding to a single cell
association or stage (Hess and Renato de Franca 2008; Chocu et al. 2012).
A mature mammalian sperm is a very small cell that has lost most of its cytoplasm during the
process of spermiogenesis. It consists of a head, neck, and tail. The tail may be further divided into a
principal piece, middle piece, and end piece. The head contains the greatly condensed nucleus and
enzymes needed to penetrate the egg’s surrounding membranes and any adhering cells. These
enzymes are located in a membrane-bounded vesicle, the acrosome, which covers the front of the
nucleus. In some species, the acrosome also contains molecular components of species-specific
recognition systems that help to ensure that spermatozoa of one species will not accidentally fertilize
an ovum of another species. The middle piece contains coiled mitochondria that have the enzymes
needed for the release of energy to the flagellum-like axial filament that powers locomotion. Because
a spermatozoon is small, it lacks extensive stores of energy, and once it begins to move, it soon exhausts
most of its potential for locomotion. Thus, spermatozoa are maintained inside a male’s reproductive
tract in an immobile state and are activated to begin swimming only by changes encountered in a
surrounding aqueous medium or by the fluids in the female’s reproductive tract. Often, the trigger that
activates spermatozoa to swim is a change in pH.
Spermatozoa of different species of vertebrates vary greatly in the sizes and in the shapes of
their heads, and sperm shape may be phylogenetically informative. For example, spermatozoa of
rodents typically have sickle-shaped heads, whereas those of most other species of mammals are
elliptical. Spermatozoa of some groups of bony fishes normally have two tails and are referred to as
biflagellate spermatozoa.
I. Illustrations
A. Examine a longitudinal section of a single lobe of grasshopper (Rhomaleum) testis. Illustrate the
different stages of spermatogenesis in various compartments or cysts.
B. Draw and label the seminiferous tubule of cross section of the rat testis showing all the
spermatogenic cells. Label the different spermatogenic cells.
C. Label the parts of the mammalian testis.

D. Indicate the different types of spermatogenic cells. Write the number of chromosomes per stage.
E. Label the parts of the human sperm.

II. QUESTIONS FOR RESEARCH. Make sure to provide your references. Use CBE or APA format.
1 Explain further the wave and cycle of the seminiferous tubule in mammalian spermatogenesis. What
is the importance of this processes?
2. What is the role of acrosome in fertilization?
3. What are the functions of interstitial cells of Leydig and Sertoli cells in males?
4. How can you distinguish a secondary spermatocyte and a spermatid? between a spermatid and
spermatozoon?
5. The primary spermatocytes’ nuclei undergo a series of changes which can be classified into stages
prophase I. What are these stages and provide distinguishing characteristic for each stage?

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ABI 3110 – Developmental Biology Laboratory

C. Label the parts of the mammalian testis.

E. Label the parts of the human sperm.

2. What is the role of acrosome in fertilization?

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