Salmon SSA
Byrnes and Gannon 1990
Lamprey LSA
Gray and Doolittle 1992
α-Fetoprotein Human
AFP Law and Dugaiczyk 1981
Rat ----
Jagodzinski et al. 1981
Mouse ----
Gorin et al. 1981
Vtamin D-binding protein Human VDP
Yang et al. 1981 Schoentgen et al. 1986
Rat ----
Cooke and David 1985
Mouse ----
Yang et al. 1990 Figure 1. Other forms of albumin and their sources
C. Relationship Between Platelets and Albumin
A study conducted by O‘Neil and Ammit 1997 showed that the release of platelet- activating factor PAF in mouse embryos was dependent upon the extracellular albumin
concentration exposed to them, with the amount of PAF released increasing as the albumin concentration increased. Small amounts of PAF were detected from the media
with no albumin added. Other types of cells have demonstrated the same dependence of PAF to albumin Benveniste and others 1972; Ludwig and others 1985. This may well
be a general phenomenon, the authors suggested.
D. Mus musculus White mice
D.1 Background on Mus musculus White mice
The laboratory mouse was derived from the common house mouse, Mus musculus. There are approximately 1750 strains, including inbred strains, hybrids,
spontaneous mutants, induced mutants, chromosomal aberrations, and wild derived. Because of the constant discovery of new mutations and the production of knockout
and transgenic mice, the number of mouse stocks and strains currently available continues to increase. In 1998, 17.2 million mice and 5.5 million rats were used at
1,200 U.S. research institutions, compared to a total of 1.2 million other species. Mice and rats together constitute approximately 90 of the total animals used for all
research purposes LAC-NUS 2007.
D.2 Validity of White Mice as Model Organisms for Blood Studies
Despite the fact that animal models have contributed a lot to our understanding of the pathological processes of various human diseases and their treatment, in many
cases the issue of reliability or validity has prevented results from animal models from being readily used to develop successful clinical treatments. Animal models
should have a genetic, naturally obtained, or induced pathological process that closely resembles the same conditions in humans. Useful models are described as
correlational, isomorphic, or homologous, but generally, the most successful are homologous models, where both the etiology and clinical features of a disease in an
animal model mirror those found in human patient. The genetic characterization, the large number of strains available, and the large
list of catalogued mutant genes provide animals suited for a number of different areas of research. Mice are easy to care for and handle, and are relatively inexpensive
compared to other species. A high reproductive performance with a large litter size and a short gestation means that many generations can be produced in a relatively
short period of time LAC-NUS 2007. Mouse platelets are known to differ from human platelets in a number of
fundamental ways, including, platelet size and platelet number. Nonetheless, studies of fundamental aspects of platelet function are quite reassuring about the similarities
between human and mouse platelets, and so there is reason to extrapolate the observations made in mice to humans.
The use of the mouse as a research animal has resulted in many scientific advancements. Much of our early understanding of the immune system was derived
from studying the mouse. The use of the mouse continues to be an important part of various
research endeavors
including aging,
embryology, cancer
induction, pharmacological and toxicological testing, and infectious diseases research LAC-
NUS 2007.
D.3 Selection of Healthy White Mice
A brief assessment of the health of the white mice should be conducted prior to performing any technical procedures. The animal should be observed for any signs of
illnesses; rough hair coat; abnormal posture; prolapsed uterus, rectum or penis; limb abnormalities;
abdominal distension;
malocclusion; dehydration;
dystocia; or
abnormal behavior.
Categories of common behaviors of mice include: maintenance behaviors grooming, eating, drinking, nesting; investigativeexploratory behaviors climbing,
digging, chewing, sniffing; and social interactions huddling together, grooming one another, scentterritorial marking, aggression, defense, sexual behavior.
D.4 Proper Maintenance, Care, and Use
Proper care, use, and humane treatment of animals used in research, testing, and education require scientific and professional judgment based on knowledge of the
needs of the animals and the special requirements of the research, testing, and educational programs.
Mice received from another site need to have adequate time to recover from shipping stress, this is known as acclimatization. The transportation of animals is
stressful and leads to physiologic changes, such as increased cortisol levels, which may potentially alter research results. The length of time required may depend on the
distance or time involved in transporting the mice but generally, a minimum of 48 hours is required for blood cortisol levels to return to baseline values. A quarantine or
holding period will allow the mice to adapt to their new surroundings and permits observation for any signs of infectious disease LAC-NUS 2007.
Proper housing and management of animal facilities are essential to animal well- being, to the quality of research data and teaching or testing programs in which
animals are used, and to the health and safety of personnel. A good management program provides the environment, housing, and care that permit animals to grow,
mature, reproduce, and maintain good health; provides for their well-being; and minimizes variations that can affect research results Guide for the Care and Use of
Laboratory Animals, National Research Council 1996. There are evidences that exist which suggest that singly housed mice may have a
compromised immune system when compared to socially housed mice. Moreover, mice developed tumors faster when individually housed than when kept in groups. In
rats, the provision of increased structural complexity has the potential to promote modifications in brain structure, physiology, and function. These changes are
mediated via increased cortical thickness, increased dendritic spine density and increased concentrations of oligodendrocytes. Based on these evidences, the
following guidelines are suggested for housing rats and mice in the laboratory:
Rats and mice benefit from being socially housed whenever possible. Mice are timid but social animals. Contact with conspecifics is important.
A mouse housed alone may become more aggressive. Adult male mice housed together may be very aggressive towards one another.
Mice benefit from being housed on nestable bedding or being provided with a suitable substrate with which to build a nest. Rats also benefit from being provided
with increased structural complexity, i.e., nest box, platforms or paper towels. Mice spend a great deal of time manipulating their bedding material, and if the
material allows they will build tunnels and nests. Bedding material provides thermal insulation, absorbs fecal and urinary wastes, and
in some instances is used for nest construction.
Material for bedding should be absorbent, not readily eaten, free of infectious agents and injurious substances, and comfortable for the animals. Bedding may consist of
paper, hardwood chips, or corncob materials. Avoid the use of materials like cotton or shredded paper in breeding cages.
D.5 Proper Labeling
Appropriate identification method for the mice is important. The choice of identification should be based on the age of the animal you wish to identify, the
number of characters you wish to include, and the duration of your experiment. Indelible markers can be used for short-term identification. Non-toxic, permanent
markers can be used to temporarily mark the fur, tail or skin of the animal. This ink, depending on the location, usually lasts 3 - 4 days without the need to remark.
Alternatively, ear punches, microchips, and tattooing are all permanent procedures. Ear tags can be long-term, but there is always a chance they can become detached
from the ear.
D.6 Proper Restraint
When attempting to restrain a mouse, it is important to select the appropriate method of restraint for the procedure you wish to perform. The restraint procedure
must offer the best access to the area requiring manipulation. Sudden, jerky moves should be avoided to decrease the likelihood of being bitten and to lessen stress to the
mouse. Restraining methods include tail restraint, forceps restraint, scruff restraint, and restraint by the use of mechanical restrainers. Tail restraint is done by grasping
the base of the tail of the mouse and forceps restraint is done by gently grasping the
mouse by the scruff of the neck. Both of these restraining methods are only used for brief restraint; for example transferring animals from cage to cage. Scruff restraint
and mechanical restrainers are used for procedures that require more than momentary restraint, such as injection or blood withdrawal.
D.7 Proper Disposal
When performing euthanasia on the mice, the procedure must be approved by the Institutional Animal Care and Use Committee IACUC. Mice may be euthanized
using the CO
2
chamber method. Compressed CO2 gas is the only recommended source of CO2 for euthanasia. Carbon dioxide generated from dry ice is unacceptable.
With an animal in a chamber, an optimal flow rate should displace 10 – 20 of the
chamber volume per minute until the mouse is unconscious. This flow rate is associated with a rapid loss of consciousness and minimal distress to the animal.
Once the mouse is unconscious, the flow rate can be increased. Gas flow should be maintained for at least 1 minute following apparent clinical death. Death should be
verified by the absence of the heartbeat, performing cervical dislocation or by perforating the diaphragm prior to proper disposal of the animal.
E. Blood Collection
