KARYOTYPE ALTERNATIVES

Tom Conley
1993 Woodrow Wilson Biology Institute

INTRODUCTION

Chromosomes are arranged into seven groups based on size and centromere location. The centromeres can be found in the middle of the chromosome (median), near one end (acrocentric) or in between these first two (submedian).

1. Group A: chromosomes 1-3 are largest with median centromere;

   Group B: chromosomes 4-5 are large with submedian centromere; Group C: chromosomes 6-12 are medium sized with submedian centromere;

   Group D: chromosomes 13-15 are medium sized with acrocentric centromere;

   Group E: chromosomes 16-18 are short with median or submedian centromere;

   Group F: chromosomes 19-20 are short with median centromere;

   Group G: chromosomes 21-22 are very short with acrocentric centromere; chromosome X is similar to group C and Y is similar to group G.

Chromosomes can also be identified by dark or light regions called bands. Bands are produced by staining the chromosomes with stains. Different stains produce different banding patterns (Levitan). See last page for chromosome grouping and banding diagram/template .

Through karyotyping, abnormalities in chromosome number and structure can be detected in the fetus. Using amniocentesis or chorionic villi sampling, the technician is able to obtain fetal cells for karyotyping. If chromosome abnormalities are discovered, a genetic counselor can educate the family about the disorder and its affects. These visible chromosome deviations are estimated to occur in 6-7 per 1000 live births and in about 1/2 of all spontaneous aborted pregnancies (Levitan). When cells have a normal number of chromosomes, they are called euploid. If on the other hand, the cells have an irregular number of chromosomes, the cells are called aneuploid. The aneuploid cells are caused by a nondisjunction during meiosis or by a nondisjunction during mitosis of the zygote. If a zygote exhibits nondisjunction after the first cleavage, then the organism is referred to as a mosaic since its cells exhibit both euploidy and aneuploidy.

Although any autosome can undergo nondisjunction, only a few of these aberrations are expressed in viable offspring. The most common autosome aberrations are Down Syndrome, Edward Syndrome and Patua Syndrome. Most other autosomal aberrations result in spontaneous abortions.

Down Syndrome or Trisomy 21 is the most common of the autosome abnormalities occurring in about 1.5 per 1,000 births. The rate for this trisomy appears to increase with maternal age. It has recently been established that the sperm can also carry this extra chromosome. Of those born with Down, 1/6 die within the first year and the average life span is 16.2 years (Levitan). The child usually has a round face, flattened nose bridge, a fold of skin above the eyes and short broad hands. The child can have an IQ between 25-74, septa defects in the heart, and a greater susceptibility to respiratory infection. Infections and heart defects increase the chance of infant death. Edward Syndrome or Trisomy 18 occurs about 1 per 6,000 births and this rate also increases with maternal age. Thirty percent of these children die within the first month and only 10% survive one year (Therman). Multiple malformations of most any organ are observed and there are severe mental, as well as developmental, handicaps. External manifestations can be an elongated skull, low-set malformed ears, deformed eyelids, possible webbed neck and unusually short sternum (Levitan).

Patua Syndrome or Trisomy 13 occurs about 1 per 15,000 births and this rate increases with maternal age. Forty five percent die within the first month, 90% by six months and less than 5% reach 3 years (Therman). External symptoms are similar to Edward's with the addition of cleft palate and/or lip, microcehphaly, polydactyl and deafness (Levitan).

Few additional autosome trisomies are seen in live births. Some have been reported for chromosome numbers 8 and 22 but these seem to be caused by nondisjunction during mitosis and thus are not expressed in all cells of the child.

Both the X and Y sex chromosomes exhibit nondisjunction. In females the most common is Trisomy X which occurs about 1 per 800 births. Symptoms range from normal physical and mental signs, to underdeveloped secondary sex characteristics (which can be treated with estrogen treatments), sterility and lowered intelligence. Most women with XXX who are fertile do not tend to pass the extra X to their offspring. This extra X chromosome tends to end up in a polar body after first meiosis. It is also possible to inherit 4 or 5 Xs. With each extra X there tends to be a corresponding increase in mental handicap, a chance of being sterile, and a lack of secondary sex characteristics (Levitan).

Turner Syndrome or X0 female occurs in about 1 per 2,700 births, is not linked to maternal age, and is thought to be a mosaic (Therman). The phenotype of this female includes short stature, short broad neck with webbing or looseness, a broad chest, lack of secondary sex characteristics and sterility. Intelligence does not seem to be affected (Levitan).

Klinefelter Syndrome or XXY occurs between 1 per 700 and 1 per 1,000 births. The man has small testes devoid of sperm, shows tendency for breast development, and has lower IQ which leads to mild mental handicap (Therman). One percent of institutionalized mentally handicapped males are XXY which is greater than what is seen in the birth rate (Levitan).

XYY males occur in the same rate of occurrence as XXY males. There are no major physical abnormalities that are indicative of this disorder. Any defects that do occur are usually confined to the genitals; such as undescended testes and defective development of seminiferous tubules. There can be some mental handicap due to the IQ range of 80-95 (Levitan).

Besides nondisjunction, chromosome mutations can be detected through karyotyping. The most common deletion, 1 per 45,000, is Cri du Chat (cat cry) Syndrome which is a deletion of a small arm of chromosome 5. During infancy the child has a cat-like cry which modifies with age. Other phenotype expressions are microcephaly, round face, low set ears, heart disease and mental handicap (Therman).

Wolf-Hirschhorn Syndrome is a deletion of a segment of chromosome 4's short arm. This affects scalp, nose, lips, palate and the penis in males. 13-Syndrome is a deletion of the long arm of chromosome 13. Severe malformations of face as well as retinoblastoma can occur (Levitan).

Two activities for karyotyping, a paper cut out method and a lab using human cells, will be described. Also if hypercard is available, refer to the American Biology Teacher, April 1992, article on "Karyotyping with HyperCard" by Jerry Jensen.

TARGET AGE/ABILITY GROUP:

1. 9th-12th grade; for any biology course. Modifications can be made so that a low level biology course or an A.P. biology course can use at least one variation.

STUDENT/CLASS TIME REQUIRED:

1. Paper Cut Outs:
   One period of 40-60 minutes if students do cut outs as homework.

HUMAN CHROMOSOME LAB:

MATERIALS:

HUMAN CHROMOSOME LAB:

SAFETY PRECAUTIONS:

Human Chromosome Lab:

TEACHERS' GUIDE FOR PREPARATION OF MATERIALS

Paper Cut Outs:

Human Chromosome Lab:

TEACHERS' OUTLINE FOR PRESENTATION OF ACTIVITY

PAPER CUT OUTS:

1. Provide a short introduction on karyotyping using background information notes and the template as an example.

2. Either provide information about the various abnormalities or assign as a library research assignment, once the student has discovered the chromosome error.

3. Hand out scrambled chromosome sheet (this could be done night before so the students could cut it out prior to class) and a sheet for students to arrange cut out chromosomes.

4. 4Students arrange and glue chromosomes to data sheet, indicate sample code, chromosome abnormality and sex of sample.

5. Students should describe the problem this subject has.

HUMAN CHROMOSOME LAB:

1. Provide a short introduction on karyotyping using background information notes and the template as an example.

2. Either provide information about the various abnormalities or assign this as a library research assignment. Students will not see any of the above mentioned abnormalities as the cells used in this kit are from the HeLa human cancer cell line that has been maintained since early 1950's.

3. The following gives the general steps for the procedure; more specific instructions are with the kit:
   A. Slides should be kept in cold methanol (40%) in a freezer or ice bath until ready for use.

   B. Remove slide and orient it vertically at a 45 degree angle.

   C. Resuspend cells in tube provided and apply 8-12 drops on the upper 3/4 of the slide from about 6-12 inches above slide (the greater the height, the better the results). This will allow the sample to "splat" onto slide.

   D. Allow slide to air dry completely.

   E. Treat slide for 1 second only in Stain #1. Repeat.

   F. Wipe off slide bottom and repeat these steps in Stain #2.

   G. Rinse with distilled water and let air dry completely.

   H. Inspect slide for good chromosome spreads.

   I. Add 2 drops of Permount on stained area of slide and cover with a #1 coverslip. Let this dry for 48-72 hours.

   J. Observe under 400X and 1000X oil immersion if available.

   K. Try to count the number of chromosomes present in 10 different cells.

4. If you have a camera attachment for a microscope, photograph some of these cells and enlarge the pictures. Make copies of this and have the students do a karyotype. Since these cells are from the HeLa human tumor line, see if they can determine which chromosome is expressing multiple copies.

STUDENT INSTRUCTIONS/HANDOUTS

PAPER CUT OUTS:

HUMAN CHROMOSOME LAB:

SOURCES OF MATERIALS

Premade karyotyping sheets available from: Carolina Biological

INFORMATION REFERENCES

The Woodrow Wilson National Fellowship Foundation webmaster@woodrow.org
CN 5281, Princeton NJ 08543-5281 Tel:(609)452-7007 Fax:(609)452-0066