BIL 250 - Genetics

#9    25 Sep 97
(complete!)
 
 ***Particularly useful portions of the text: pp. 221-234, 103-104***
 
Cytogenetics (con't)
 
Numbers of Chromosomes

    While fission and fusion can change chromosome number, these events are quite rare. A more common event that occurs affecting chromosome number involves a type of meiotic error called non-disjunction. Non-disjunction describes that situation when chromatids are not equally distributed among daughter cells during meiosis. Resulting gametes will have too many or too few chromosomes. If these gametes become part of a zygote, the resultant individual is said to be aneuploid. Sometimes such individuals are viable, but often there are developmental problems that result leading to individuals that are different. Many cases, however, result in zygotes that do not complete development or do not live long. If a gamete with an extra chromosome combines with a normal chromosome to produce a zygote, that zygote is said to be trisomic: such zygotes have three of one chromosome rather than the usual complement of two. Trisomy-21, for example, is a fairly common situation in humans, resulting in what is termed Down's Syndrome. Conversely, monosomy refers to a zygote with a missing chromosome: it has one of a chromosome rater than the normal complement of two. Most cases of monosomy result in zygotes that do not fully develop. Look over the beginning of Ch. 9 to see some other examples of these phenomena.
 

Sex Determination, Sex-influenced Traits, and Sex-linked Traits
 

    Until now, we have concerned ourselves only with genes that largely conform to the basic Mendelian Model (diploidy, normal segregation). This holds for genes and chromosomes comprising most of the genomes of organisms. These chromosomes are termed autosomes. The other type of chromosomes are called sex chromsomes: not surprisingly, this is becauase have something to do with the sex of individuals... We will continue our genetical study by first examining a few sex determination systems, then look are traits that are associated with or affected by genes on sex chromosomes: sex-influenced traits, sex-limited traits, and sex-linked traits. Along the way, we will take a brief look at dosage compensation.
    The first suggestion that chromsome types may be associated with sex determination came from cytological studies of in some insects. In the late 1800s and early 1900s, work by Henking and by E. B Wilson on a species where males had an odd number of chromosomes (13) while females had an even number (14) resulted in an understanding of the details of this system. It was found by careful microscopic observation that all female gametes had 7 chromsomes, whereas males produced equal numbers of gametes with 6 and 7 chromsomes. Further experimental work allowed construction of the details of what is now known as the XO sex determination system, found in many species of insects. Females have 6 pairs of autosomes and 1 pair of sex chromsomes. All female gametes have 6 autosomes and 1 sex chromsome. Males, on the other hand, have 6 pairs of autosomes and a single sex chromosome. This sex chromsome has no other chromsome to pair with during meiosis, so half of gametes produced receive a sex chromsomes and half do not. Therefore males produce two distinct types of gametes, and are referred to as the heterogametic sex in this system. (Females are referred to as the homogametic sex.) It is therefore the male gamete that determines the sex of the zygote: if the sperm has a sex chromsome, the individual is a female, and vice versa. Henking first referred to the sex chromsomes as "X-bodies": this resulted in the sex chromsomes in this system being called X-chromsomes.
    Although this was the first sex determination system discovered, it is not very common. A more common system is called the XY sex determination system. The cytogenetic characteristics are similar, except that there is a distinct chromosome type (called the Y-chromsome) that pairs with the X-chromosome during meiosis. Again, males are heterogametic and females are homogametic. However, is this system all male gametes have the same number of chromosomes, but they differ in whether they contain an X-chromosome or a Y-chromosome. For example, a species with a diploid number of 20 would have 9 pairs of autosomes and 1 pair of sex chromosomes. Females would have 18 autosomes and 2 X-chromosomes, whereas males would have 18 autosomes, 1 X-chromsome, and 1 Y-chromosome.
    A third type of chromosomal sex determination, found in birds, moths, and some fish, is similar except that the famales are heterogametic and males are homogametic. This is called the ZW system: males are ZZ and females are ZW.
    Although the chromosomal characteristics of species with XY sex determination are similar, there are substantial differences in the mechanisms that actually determine sex. We will briefly examine two of the more well-studies cases: 1) The Genetic Balance System studied extensively in Drosophila, and 2) the mammalian XY system.
    In Drosophila, XY individuals are male and XX individuals are female. (All individuals have 3 pairs of autosomes.) However, the presence/absence of the Y-chromsome has no effect on sex: it is the number of X-chromsomes relative to the number of sets of autosomes that determines the sex of individuals. This system became understood after extensive work by C. B. Bridges involving careful examination of chromosomal complement of individuals that were the result of non-disjunct gametes. In the following discourse, we will use "A" to refer to the set of autosomes (3 pairs). Normal males are AAXY and normal females are AAXX. Male gametes are normally either AX or AY. However, when non-disjunction occurs, there are many possibilites: A, AX, AY, AXX, AXY, AAX, AAXX, etc. The gamete will have various combinations of sex chromsomes and autosomes depending on the exact nature of the non-disjunction. Similarly, females gametes are normally AX, but with non-disjunction may be A, AXX, AA, AAXX. When abnormal gametes combine, individuals with abnormal combinations of autosomes and sex chromsomes will result. Detailed work led to the following obervations (among many):

                            Genotype        Result
                            AAXX            normal female
                            AAXY            normal male

                            AAAX            male (infertile)
                            AAAAXX      male (polyploid)
                            AAXXX        female (infertile)
                            AAAAXXX   intersex

Bridges and co-workers evaluated these and other data and determined that it was the ratio of autosomal sets to X-chromsomes that determined the sex. This is summarized below:

                            Ratio                      Sex
                            X/A <= 0.5            male
                            X/A >= 1.0            female
                            0.5 < X/A < 1.0     intersex

Because it is a balance of autosomes and sex chromosomes that determines sex, this became known as the Genetic Balance System.

    Mammals have XY sex determination, but have a very different underlying mechanism. Again, this system was elucidated by examination of individuals with unusually complements of chromsomes because they came from non-disjunct gametes. The following are some examples (here, we ignore autosomal sets):

                            Genotype                   Sex
                            XX                            male
                            XY                            female

                            XXX                         female
                            XXY                         male
                            XYY                         male

These and and other data indicated that it is the presence of the Y-chromosome that determines sex. The hypothesis is that a gene on the Y-chromosome causes embryonic gonadal tissue to develop into testes: in the absence of this gene, the tissue develops into ovaries. This gene produces what is called testis determining factor (TDF). Recent work has provided substantial evidence for this hypothesis:
    1) Some individuals that are XX are male and some that are XY are female. Careful karyological examination indictes that in these individuals there has been a change in the sex chromosome. In the first case, a small part of the Y-chromosome has been deleted. In the latter, there has been a translocation of the same portion of the Y-chromosome onto one of the X-chromosomes.
    2) Work on transgenic mice indicates that the injection of a specific gene region from that same small part of the Y-chromosome into a normal XX zygote at an early stage of development results in a normal male. This region (in mice) has been termed the SRY (sex-determining region).
 

Dosage Compensation

    You may have noticed that in species with XY sex determination, males have 1 X-chromsome and females have 2 X-chromosomes. Does this mean that females get a double dose of genes on the X-chromosome? The answer is no: there is a phenomenon called dosage compensation whereby one of the X-chromsomes is inactivated in all cells in females. This inactivation is random, and it is not the same in all cells in the body. Females are therefore mosaics with respect to genes on the X-chromosome: some tissues have the paternal X-chrosomes inactivated and others have the maternal X-chromosome inactivated. How do we know this? This idea is enbodied in the Lyon Hypothesis, originally formulated by Dr. Mary Lyon. It has been substantiated in several ways. One way involves the interpretation of banding patterns that are visible as a result of protein electrophoresis. The enzyme G-6-PDH occurs on the X-chromosome. It is a dimeric enzyme, meaning that the functioning enzyme is comprised of two identical subunits, each the result of translation of the gene. These subunits randomly combine in the cytoplasm to form functioning enzyme molecules. Females that are heterozygous can produce two different type of subunits, that can combine in 3 different ways. If we call these subunits A and B, then it is possible for the functioning enzyme to be AA (2 A subunits), AB, or BB. Electrophoretically, these are distinguishable. If an individual has only one active gene in each cell, then only one type of subunit is present. Therefore, they could only exhibit AA and BB enymes. If, on the other hand, the same X-chromosome was inactivated in all cells, then only AA or BB could be produced. If there was no inactivation, then all 3 types could be found. Electrophoretic results indicate support for the Lyon Hypothesis: in tissue preparations from some females, both AA and BB were found, but no AB types were found.
    At this point it is useful to interject a bit more about sex determination. Although we have seen 3 different types of chromosomal sex determination systems, there are many organisms where the environmental conditions determine sex. Many reptiles and fish have there sex determined by the temperature experienced during a specific critical period of development. Other species, such as.....(drum roll...) Daphnia have another type of environmental sex determination. In this system, it is the general environmental condition that determines sex: if food is abundant and the population is not crowded, only females are produced. If the population is becoming crowded and food is scarce, males are produced.
    However, even though these two systems are not chromsomal, and are termed ESD (environmental sex determination) it is important to not that there are genes that affect sex. Genotypes differ in there response to environmental stimuli in terms of the sex ratio on their offspring. Therefore, rather that dichotomizing sex determination into Genetic vs. Environmental, it is probably better to dichotomize between Chromosomal and Non-chromosomal, and recognize that the latter case consists of a wide variey of mechanisms with differing degrees of genetic contribution.

Sex-influenced traits
    Now that we have learned something about sex determination, let's turn our attention to traits for which the sex of an individual has an important effect on the expression of a trait. The first type of trait are sex-influenced traits. These are traits in which the expression of the trait can occur in both sexes, but that the relationship between genotype and phenotype will be affected by sex. For example, in British sheep, there is a locus that essentially determines the expression of horns. There is an h+ allele associated with the "horned" phenotype, and an h allele associated with the "hornless" phenotype. In males, h+ is completely dominant to h, whereas in females the converse is true. Therefore, an h+h+ genotype is always horned, and an hh genotype is always hornless. The h+h genotype is horned in males, but hornless in females. Because there are many example such as this one where it is the dominance relationships between the alleles that is affected by sex, such traits are often referred to as being instances of sex-influenced domiinance.
    Sex-limited traits

    The next category of traits are those for which the expression is limited to one or the other sex. An example of this (discussed in class) is a class of lineages of Daphnia pulex, which all contain a mutation which is a sex-limited meiosis suppressor. There is a dominant allele at a locus that, if present, suppresses meiosis in females: it has no effect on males. Therefore, the trait ("meiosis suppression" in this case) is limited to one sex.