WORKSHEET

Zebrafish Development: tool for studying Genetics

Introduction

In this module, students will learn how and why biologists use model organisms,
specifically zebrafish, to study human development and diseases, and those caused
by mutations. In part one, students will study the development of zebrafish and why
they are used for human research. In part two, students will identify and study
mutations by using pictures of zebrafish experiments.

Learning Objectives

At completion of this activity, learner will:

1. Describe the developmental stages of zebrafish and why they are used for human
research.
2. Identify common mutations of zebrafish.

Material
Zebrafish Development _hand-outs

Method
1. Present the hand-out of zebrafish in research.
2. Pass out activity worksheets (this form).
3. Have students work on the “spot the difference” activity.

Questions and student’s answer

1. Image 1 shows an eyeless mutant that is linked to eye development. A) What’s
the difference? B) Why is this relevant to us? Discuss at the gene level.
* Embryo B has no eye.
* Zebrafish and humans have a network of proteins, called the Wnt pathway,
connected with the creation and growth of embryos. Eyeless embryos, such
as the one shown in the image, can occur via gene mutations encoding the
proteins in the Wnt pathway. Identifying the genes involved in those pathways
will help us understand the role of the Wnt pathway in early brain and eye
development.

2. Image 2 shows a pigment mutant that is linked to skin color. A) What’s the
difference? B) Why is this relevant to us? C) Can you see the difference between

Professor: MTN Cabasan 1

 the melanosomes in the darker striped fish and golden fish in the picture?
Discuss at the gene level.
*Fish B is a little lighter and golden in color than fish A.
*Lighter skin colors in human is linked to a reduced number, size and density
of melanosomes. These are structures found in skin cells that contain a dark
pigment called melanin. People with darker skin have lots of large and tightly-
packed melanosomes in the darker striped fish and golden fish in the picture.
The difference in color has been linked to a gene called SLC24A5 which is
present in both zebrafish and human.
3. Image 3 shows a nebulin mutation which is linked to congenital nemaline
myopathy. A) What’s the difference? B) Why is this relevant to us? Discuss at the
gene level.
*The image shows two young zebrafish known as fry. The body of zebrafish B
is curved. If you look closely you’ll see that its mouth is open. This is because
unable to fully close its mouth as its muscle are too weak.
*Zebrafish fry B is showing symptoms of a condition called nemaline
myopathy. This is a genetic disease where muscles fibers do not form and
function properly. This leads to poor muscle tone and weak muscles. Infants
with this condition usually have problems breathing and feeding as their
muscles are not strong enough to support the body. This can also lead to
skeletal problem such as curvature of the spine (Scoliosis).
Genes that are associated in this kind of condition is both present in
zebrafish and human genomes. Observing nemaline myopathy in zebrafish,
both its physical and genetic changes can help us understand more about this
condition and can lead us to better treatments for the disease.

4. Image 4 shows mutants that cannot form red blood cells. A) What’s the
difference? B) Why is this relevant to us? Discuss at the gene level.
*The zebrafish embryos in picture B look paler and are not stained red.

* Red blood cell formation and iron uptake has been found to be linked to a
gene called ARHGEF3 that is found in both human and zebrafish. This gene
encodes exchange factor which activates other genes along a cell signaling
pathway. When the function of this gene and its target genes is blocked it
leads to a severe decrease in mature red blood cells. This type of research
can give us a better understanding of the function of the genes and the roles
that they play in iron metabolism and the development of red blood cells and
blood vessels.

Professor: MTN Cabasan 2

5. A) What determines the development of the body parts of the zebra fish? B) Why
do the zebra fish develop these mutations? Relate your answer to the
Developmental Genetics lecture (Topic 9, VLE).
*As zebrafish eggs fertilized and develop outside the mother’s body it is an
ideal model organism for studying early development. Zebrafish have a similar
genetic structure to humans that’s why they are widely used in research due
to their numerous beneficial properties. They are also very cheap; however,
they do require more space than the other model organisms such as flies.
This model is also vertebrate, which gives it am advantage over other models.
Zebrafish embryos can also be genetically modified. Zebrafish share 70% of
their genes with humans, and 84% of disease-related genes have zebrafish
counterpart. Zebrafish genome has been fully sequenced and over 140, 000
genes have been mutated to study their function in development and disease,

Professor: MTN Cabasan 3