TPC NEWS Fall 1988 Vol.7 No.1 (Whole Number 14)
(English Summary)
Page-3
Essay: TPC, the past ten
years and the future
Our Tsukuba Primate Center (TPC) was established about ten years ago.
A decade is a suitable time to look back on and recall our history, as
well as to imagine the future. But
I am going to retire from TPC under the age limit, in less than two years.
This short essay may be said to be something like my 'will' to the
younger people, even though the time is too early yet.
TPC's purposes are to breed and rear nonhuman primates of good quality
as laboratory animals, and to carry out basic, technological studies
related to the breeding of monkeys. However,
Dr. H. Fukumi0, who was the director of the National Institute of Health in
Japan at the time of TPC's establishment, once advised us in the TPC brochure
not only to get good results in breeding, but also to achieve development in
basic research. Professor K.
Yamanouchi of The Institute of Medical Science, The University of Tokyo also
contributed to TPC NEWS ( Vol.5, No.2,
1986), an article expressing his expectation that TPC should play an important
part in the study of laboratory primates like that being played by the The
Jackson Laboratory in the U.S.A. in the study of laboratory mice.
So far, our efforts have resulted in establishing our own breeding
system, by which laboratory primates of good quality can be bred successively.
Now we are able to define genetic profiles
of each monkey and its family relations by various phenotypic
characteristics such as blood types, serum proteins, lymphocyte defined
antigens, mitochondrial DNA and so on.
This is very significant from the view point of the preservation of
genetic resources of wild life. And
this also means that now we have the potential ability to cope with the needs of
almost all biomedical facilities concerned with primate experiments nationwide,
if conditions such as budget, manpower and systems permit.
On the other hand, what is the present state of our research activities?
I have often mentioned TPC's research policy which I have followed
personally and have asked our young researchers to observe (TPC
News Vol.2, No.3, p.3, 1983; J.Med.
Primatol., Vol.14, No.2, p.75-89,1985; Reichorui-kenkyu
(Primate Research) Vol.1, No.1, .31-38, 1985; TPC
News, Vol.5 No.2, p.3, 1987); Jikken-dobutsu
(Experimental Animals) Vol.36, No.4, p.367-379, 1987).
I heartily hope that TPC's researchers do their best and accomplish
something useful for society through their studies, according to the policy.
By the way, we are in a very blessed position to carry out research,
because every monkey kept at TPC is a research subject as well as a research
material. But the number of our
researchers is too small to cover completely the very wide research fields of
primatology and primate medicine, and to use
these monkeys fully. Some parts of
the studies are conducted as cooperative studies by visiting researchers
from other institutes or universities.
Moreover, the recent budget cuts threaten TPC's financial base.
It is impossible to improve even basic research unless this situation is
drastically reformed.
Recently, a movement asking for the establisment of nationwide primate
centers, of which TPC will be the core, has developed among researchers in
industry, university and government. The Committee for Future Reformation of NIH,
Japan concluded last year that TPC should be reorganized.
Furthermore, the Council of Science and Technology for National Health
and Welfare called for establishing a stable laboratory primate supply system.
Judging from these situations, I fully realize that we should do our duty
and take new steps forward as
researchers based on a fundamental reform in TPC's organization.
We must tread a thorny path, but now I recall a proverb,
"Where there is a will, there is a way."
Page-4
Breeding Topics: Time
of deliveries in cynomolgus monkeys under laboratory conditions
We described in the last issue of TPC NEWS that deliveries of the
cynomolgus monkeys kept at TPC would most frequently occur between 19:00 and
20:00. In this issue we present
more detailed data on the time of deliveries.
Four TV cameras were set in front of eight monkey cages, and recorded
full-term monkeys' behaviors for fifteen seconds per minute around the predicted
time of delivery.
The results obtained are shown in the Figure.
Eighty-nine cases out of 122 deliveries (89%) occurred for the
period of 20:00-02:00. There
was no significant difference between wild-originated monkeys and colony-bred
ones, except for the two cases of wild-originated monkeys observed in the
daytime.
Page-5
A
survey on antibody against simian cytomegalo virus (CMV) in colony-
bred cynomolgus monkeys
We carried out surveys on antibody against simian cytomegalo virus (CMV)
from June, 1986 through December, 1987, using an indirect immunofluorescent
method.
The results obtained are shown in the two Tables.
Page-6
A Special Article: Type II collagen-induced arthritis in cynomolgus monkeys as
an experimental model for rheumatoid arthritis in humans
The author of this article, Dr. Terato, describes why he used cynomolgus
monkeys as an experimental model for rheumatoid arthritis in human, presenting
some data of his study.
The role of collagen in the pathogenesis of rheumatoid arthritis (RA)in
human was suggested for the first time by Steffen et al, who reported the
presence of anti-collagen antibodies in the sera of RA patients.
However the findings that living bodies are composed of various types of
collagen, and the discovery of antibodies against different types of collagen in
RA sera, complicated the problem, giving a clue for the study on immunization
with collagen.
In 1975-77, Dr. D. E. Trenthan of the University of Tennessee discovered
that the mice immunized with bovine type II collagen developed polyarthritis.
This was the beginning of the study of type II collagen induced arthritis
(CIA). But arthritis experimentally
induced by immunization with autologous or heterologous type II collagen in
rodents was not generally accepted as a model for rheumatoid arthritis in
humans, because of the difference in etiological and histo-pathological
features. For example, one
criticism of the use of CIA in rodents as a model of RA was the absence of
sex-linked differences in susceptibility which was found in humans.
Female mice were more resistant to CIA than males, unlike humans in which
rheumatoid arthritis is more prevalent in women.
Therefore, it was a critical problem which animal species we have to use
as an experimental animal for rheumatoid arthritis in human. The animal used had
to be closer to human in the phylogenetic distance and to be easily obtainable. We decided to use cynomolgus monkeys.
We immunized five male and ten female cynomolgus monkeys with chick type
II collagen (CII) to clarify the role of sex-linked factors in the pathogenesis
of CIA in monkeys, observing the development of rheumatoid arthritis as well as
examining the immune response to CII and monkey type II collagen (MkII).
The data and findings we obtained are shown in the Figs. 2, 3, 4, and 5.
They are demonstrating that susceptibility to CIA in monkeys is
sex-linked, that is female monkeys are more susceptible than males, and directly
related to the production of autoantibodies to MkII.
Furthermore these data demonstrate the establishment of CIA in cynomolgus
monkeys as a model of human rheumatoid arthritis ,and imply that susceptibility
and resistance of cynomolgus monkeys to CIA may be regulated by an epitope-specific
suppressor mechanism which controls the production of cross-reactive antibodies.
Page-8
The
Fourth General Assembly of Japan Primate Society
The Fourth General Assembly of the Primate
Society of japan was successfully held at Tsukuba University in Tsukuba city on
July 7th to 9th, 1988. The
number of its participants was over 300.
Page-9
A Comment on My Research:
Gene therapy and nonhuman primates
Recent progress of the technology in genetic engineering and molecular
biology made it possible to detect genetic diseases before their onset.
And, the gene therapy, in which normal DNA sequences are transferred to
the patients in order to cure them radically, has become to attract public
attention. It also has thrown many
topics which should be fully discussed: Whether
its application to the patients is the best treatment? What will happen if it is applied to the germ cells?
And so on. This paper
describes the present status of the research in gene therapy, together with the
significance and the possibility of the somatic gene therapy in nonhuman
primates.
Page-12
A report on the institutes for laboratory
animal science in People's Republic of China
Dr. Honjo, Director of TPC, and Dr. Yoshida, Senior Scientist of TPC,
visited People's Republic of China in March this year(1988) for the
development of the Collaborative Program on Laboratory Primates between Japan
and China.
The institutes introduced in his report are:
1.
Institute of Medical Laboratory Animal, Chinese Academy of Medical
Science.
2.
Laboratory Animal Center, Academy of Military Medical Science.
3.
Yunnan National Laboratory Primate Center of China.
4. Medical Primate Center
of China Institute of Medical Biology, Chinese Academy of Medical Science.
5.
Kunming Institute of Zoology, Academia Sinica.
6.
South China Institute of Endangered Animals.
7.
The Laboratory Animals Breeding Experimental Farm, Shunde Guangdong.
Both Drs. Honjo and Yoshida were deeply impressed with the earnest
devotion of Chinese researchers to the study of laboratory primates and with the
developing facilities in China.
Page-14
Two notes about new species of primates
(1)
Discovery
A British zoologist, Mike Harrison of Edinburgh University, found a new
species of monkey in the rainforests of Gabon in central Africa.
It was named "solatus", meaning sunstruck, because the tail was
bright yellow-orange from half way along to the tip. ------From
"New Scientist", 23 June, 1988.
(2)
Confirmation
The lemur first observed in 1985 by Bernard Meier and Yves Rumpler in the
Hanomafana region of Madagascar was confirmed to be a new species.
------From "Nature", Vol.333, 19 May, 1988.
Page-15
A Report from the Front of Animal Care and Feeding: Complete physical
checkup
"How are you, our readers?"
Nowadays, adult diseases are serious problems in our society.
So, it is needed for the aged or middle-aged to enter hospital for a
thorough physical checkup once or twice a year.
Every monkey of TPC, like us, undergoes a complete health checkup once
about a year. Today I will show you
the whole aspect of this health check.
The
complete physical checkup is one of the most significant works in TPC.
It is carried out on every Tuesday so
as to grasp each monkey's health condition and the prevalence of some microbes
in our monkey colonies. The items
of this health checkup are as follows: tuberculin tests, weight measuring, and
dental, hematological, biochemical, bacteriological, virological and
ophthalmoscopic examinations. A
special team composed of nine members selected from our laboratoy staffs and
animal technicians conducts these examinations. Monkeys undergoing the
examinations are listed up by computer.
Today's monkey catcher is Mr.
O.. He catches monkeys in the cage with a catching net.
Another Mr. O shoots anesthetic in the thigh of the monkeys.
The anesthetized monkeys are transferred to the next step: tuberculin
test and weight measurement. Mr. F.
records the monkeys' weight, and inoculates them 1 ml of tuberculin into the
left upper eyelid. The reaction of
this test is examined 48 hours later. Fortunately,
no positive case has been found so far. The
number of the monkeys into which Mr. F. has inoculated tuberculin may be listed
up in Guiness Book of Record!?
Then the monkeys are subjected to a strict body inspection.
Today, Mr. T. is in charge of this body inspection.
He checks dental, oral, nutritional conditions and trauma.
Nutritional condition is divided into four degrees, -, -, -, and -.
The thickness of the abdominal subcutaneous fat is also recorded.
We observe the body condition of our monkeys everyday without palpating,
but today the inspection is done more strictly from the the top of the head to
the tip of the tail. By this
inspection we once found an
abnormal tumor in the abdomen of a monkey and
it was revealed later to be ovarian cystoma.
Next, the monkeys are transferred onto the table for blood taking.
Mr. N. is taking 5 ml blood from each monkey weighing less than 1.5 kg,
and 7 ml from pthose weighing over 1.5 kg.
Of which 1.5 ml blood is poured into a tube containing anticoagulant for
biochemical examinations, and the remaining blood is stocked in the serum bank.
The final stage of today's work is the examination of ocular fundus by
Mr. S.. He is an expert of this
part. He takes pictures of the
ocular fundus with an ophthalmoscope.
One drop of mydriatic is instilled into each eye of the monkeys in
advance. Now, he has more than
30,000 pictures of the ocular fundus of monkeys.
The number must be also a world record.
His study on the ocular fundus has been presented in this TPC NEWS before
(Vol.3, No.1, 1984). Well, it
is the time of recovery from anesthesia. The
monkeys are returned to their home cages by the two Mr. 0.s
I mentioned before. As you
know, whether today's work takes time depends on the cooperation of team
members. Today's team is a very
good model which showis a fine team-work in TPC.
Aliquot smples of blood taken in the morning are analyzed with an
auto-analyzer by Mr. Y. and Mrs. O. in the afternoon.
The analyzing procedures and data also have been presented before in TPC
News (Vol. 4, No. 2, 1985). If we
note an abnormal figure, we recheck the animal showing it.
Moreover, the blood samples are examined virologically.
Antibodies against measles, SV5, HSV and sometimes retroviruses are
tested. So far, we have not found
any positive case against measles and SV5 in our colony-bred animals.
This year, TPC celebrates its tenth anniversary.
The items of this complete health checkup were reconsidered, and some of
them such as bacteriological examination of feces, antibody test against SV5,
and etc., have been omitted. We are always searching an effective way being
suited to the present state and the future of TPC.
The complete health checkup conducted for other primate species kept at
TPC, African green monkeys, common squirrel monkeys and tamarin, is basically
the same as that for cynomolgus monkeys. However,
some items of biochemical or virological examinations are different.
Characteristic viruses checked differ in those species.
I hope that the data and experience we obtained through this complete
health checkup will be useful for the people working with monkeys, and that the
monkeys reared carefully like this should never be used in vain.
Page-18
A Memory of a Trainee: Changes in the interactive behavior between mother and infant
colony-bred monkeys for six months after birth
A trainee from Tsukuba University studied for a year on the changes of
the interactive behavior between mother and colony-bred infant cynomolgus
monkeys at TPC.
She observed four mother-infant pairs, of which two infants were male,
recording their behaviors based on the behavioral items she had previously
selected. The
results obtained are shown in the three Figs. on page 19.
Page-20
Case Report:
Diverticulosis of the colon in a cynomolgus monkey
Diverticulosis of the colon is comparatively popular in man. However, the
most part of its pathogenesis is yet unknown.
Very few animal models of this disease are established.
There seems no case report on this disease in cynomolgus monkeys.
Bookus et al. have reported that the human patient of this disease was
often complicated with inguinal hernia. A
case we found in a cynomolgus monkey also concurred with herniation.
It may become a significant animal model to clarify the cause and
pathogenesis of this disease.
Case:
A wild-originated male cynomolgus monkey imported from Malaysia.
His age estimated on arrival was five or more years.
After importation he had been housed for eight years at TPC.
Commercially prepared monkey diets and fresh fruits were given.
There were no significant
findings on stool properties. His
nutritional condition was good at the time of autopsy.
Hernia was recognized in the left inguinal part
and the lower abdomen.
Histopathological
findings:
Three large diverticula were found on the surface of the serous membrane.
Two of them were 1.1 0.6 cm and 0.6 0.3 cm in size, respectively, and
papillary in shape. Dark brown
contents were seen through. Minute diverticula upheaving several millimeters were dotted,
too. All of these minute
diverticula were formed on the
surface of the serous membrane between the mesentery and its opposite side. The thickness of the muscle layer around the diverticula were
normal or were markedly thinned. Both
the large and minute diverticula were observed at the point where blood vessels
run through from the serous membrane to the circular muscle.
These diverticula were formed with lamina mucosa and a layer of lamina
propria being pressed and shrunk. The
lamina propria was infiltrated with a small number of small round cells,
eosinophilic leukocytes and histocytes ingesting brown dyes.
Tunica muscularis was vanished at the cervices of those diverticula. This case was diagnosed pseudodiverticulosis of the colon by
these findings.
Diverticulosis seems to be, in its entity, herniation.
Therefore the multiple herniation often seen in diverticulosis is not
always a coincidence. Constitution
and age are considered as the factors relating to the occurrence of this
disease.
Page-21
Mothering response of colony-bred female monkeys
An animal-technician is describing how
to teach mothering response to colony-bred female cynomolgus monkeys who can not
nurse well their infants.
At TPC, parturitions by colony-bred females have exceeded half in number.
But mother monkeys who experience parturition for the first time
sometimes fail to nurse or take care of their own infants.
Some of them hold their babies upside down, or show no interest in
nursing. Some measurement to teach
them how to nurse their babies are required.
Firstly, mother monkeys being no good at nursing are taught the way how
to hold babies by animal-technicians. And
they are housed in the same cage with other experienced mother monkeys for a
certain period. A special room
where pregnant monkeys of the last gestation period and mother monkeys of
nursing period are kept together is provided in the building for breeding.
Monkeys who become pregnant for the first time are transferred into this
room from the early period of gestation to learn how to deliver or nurse by
seeing experienced mother and other pregnant monkeys. Monkeys can learn by seeing like us.
Page-21
Mating
potency test for colony-bred male monkeys
The mating potency test we adopt at TPC is as follows:
We select healthy candidate breeders from sexually matured male monkeys.
Their canine teeth are cut not to hurt female monkeys. A male candidate breeder is housed together with a
nuriparus colony-bred female who has regular menstrual cycles, for 14 days after
7days the first day of menstruation. Vaginal
swabs are taken from the female every day to confirm the presence of sperm.
Thirty-five days after the end of the 14-day period, the female undergoes
diagnosis of conception. A male candidate is tested by turns with four females
in this procedure.
We select fertile breeders from these candidates. The male monkeys, whose
sperms are not observed in the vagina of his counterpart, or who attack females,
are not used as breeders.
The results obtained from the mating potency test for 26 male candidates
are as follows:
14 males (54%) fertilized female.
7 males (27 %) were confirmed only their sperms without producing
pregnant case.
5 males have no sperm and did not fertilize.
We judged that 80% of the
tested male candidates were fertilizable.