Evan McCarvill

Biology 3042
Dr. Daniel Ruzzante
November 29th, 2007

What we know about balancing selection and genetic drift and their relative roles in
the maintenance of MHC diversity in fish

Introduction:
The question I seek to answer is what relative roles are played between genetic drift
and balancing selection in the generation and maintenance of the genetic variation in the
major histocompatibility complex in fish. I will argue, based on my reviews of the
literature, that balancing selection is the primary cause of the great genetic variability
seen in the major histocompatibility complex and that many factors can influence the
degree of selection pressure that it experiences. However, I will also argue that random
genetic drift can, at times, interrupt, or overwhelm these processes. Balancing selection
generates and maintains major histocompatibility complex diversity through selection
pressures imposed by pathogens and also by disassortative mating to prevent inbreeding.
Genetic drift, on the other hand, is a short-term, neutral process that often occurs as a
result of low population size, bottlenecks, or species-specific behavior.

What is the MHC:

The major histocompatibility complex (MHC) is a gene cluster found in all jawed
vertebrates that produces cell-surface proteins that are found on all cells of the body
(Wikipedia). The MHC is instrumental for the recognition of foreign molecules, such as
those of pathogens, by the immune system and distinguishing them from native
molecules. The cell-surface proteins consist of a basic structural portion of
comparatively little variability and a much more highly variable portion that functions as
a recognition site for foreign molecules. The MHC genes code these cell-surface
proteins, which serve to present short peptides of foreign molecules to T-cells. This in
turn, results in an immune response to these recognized foreign molecules. Since the
high polymorphism of the MHC proteins is found primarily at the peptide recognition
regions of the molecules, the variability of the MHC is, not surprisingly, associated with
greater immunocompetence, and therefore with fitness, in a number of vertebrate species
(Lohm et al., 2002; Penn et al., 2002). Because the MHC plays such a significant role in
a vertebrate’s immune response, the maintenance of the diversity seen in this genetic
system is thought to be greatly influenced by pathogen-driven selection pressures
(reviewed in Bernatchez and Landrey, 2003).

Mechanisms of selection:
There are two prominent hypotheses that could explain the generation and
maintenance of the high MHC polymorphism that is being observed. They refer to the
pathogen driven mechanisms of balancing selection. They are the heterozygote
advantage hypothesis and the rare allele hypothesis (Hedrick and Kim, 2000).
The heterozygote advantage hypothesis supposes that individuals that are
heterozygous for genes of the MHC are selected for because they posses a broader variety
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of alleles and thus can recognize a wider range of pathogenic antigens. With the
heterozygotes being favored in a population, certain alleles are stabilized at certain
frequencies without any allele being lost or fixed. This creates a stabilization of a variety
of allele frequencies, resulting in the maintenance of the observed high MHC variability.
As yet however, there is fairly little supporting evidence from population surveys and
experimental infections supporting this hypothesis (Arkush et al., 2002).
The rare-allele hypothesis, on the other hand, supposes that by virtue of mutation or
gene conversion, a rare allele is generated that grants superior resistance to a pathogen
than is provided by other alleles. This rare allele is thus favored by selection and quickly
spreads through the population. However, it will not become fixed, because once it
becomes common, it increases selection pressure on the parasites or pathogens to evade
recognition by this allele (reviewed in Jeffry and Bangham, 2000). The time lag between
these opposing evolutionary responses would lead to a cycling of fitness values for
different genotypes in both hosts and pathogens, thus resulting in the observed
heterozygosity and variability (reviewed in Bernatchez and Landry, 2003). Many recent
studies have found correlations between pathogen resistance and specific alleles of the
MHC, providing evidence for this hypothesis (Meyer-Lucht and Sommer, 2005).
In accordance with these hypotheses, there have indeed been a few reports of
pathogen resistance being enhanced among MHC heterozygotes, which would have
increased MHC variability (e.g. Penn. et al., 2002). This is also supported by evidence of
increased pathogen susceptibility among inbred individuals, where of course, MHC
variation would be reduced (e.g. Coltman et al., 1999). There have been associations
found in Chinook salmon between MHC heterozygosity and infectious diseases in free-
ranging animals under natural conditions (Oncorhychus tsawytscha, Arkush et al., 2002).
There has also been evidence that MHC diversity in striped mice (Rhabdomys pumilio) is
maintained through pathogen driven selection by both heterozygosity advantage and also
by selection of a specific allele, although it is not yet known which mechanism is more
important for balancing selection (Froeschke and Sommer, 2005). However, it should be
noted that there is no reason to believe that the two possible mechanisms of selection are
mutually exclusive.
A population with higher exposure to pathogens would, of course, be under higher
selection pressure and so, by virtue of Darwinian selection, would show greater
variability in the MHC. Low MHC variation may be explained by low exposure to
pathogens, and lower selection pressures (Hambuch and Lacey, 2002). Natural guppy
populations have shown a good deal of variation in terms of parasite fauna and also
immunocompetence (Cable and van Oosterhout, 2006). This would make sense indeed,
if different parasite or pathogen species provided differing selection pressures. Another
study conducted by Oosterhout et al in 2006, on Trinidadian guppies, showed that the
strength of selection appears to be proportional to the frequency and gravity of disease
epidemics, as one might expect, and that the high selection coefficients tend to occur in
small and genetically isolated populations. Not only might genetic and geographic
isolation render a population more susceptible to novel pathogens, but it also might result
in a reduction of large predators, and other selection pressures. This would render the
effects of parasite mediated selection more evident (Osterhout, et al., 2006). Although a
small population would be expected to show low genetic variation due to drift, a
population of guppies which was small in size over the long term has nevertheless been
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shown with MHC variability comparable to that of a much larger population, due to the
high selection pressure being imposed by parasites and pathogens (Osterhout, et al.,
2006).
It is also thought that reproductive mechanisms, such as disassortative mating, play a
role in some species to maintain MHC diversity (Apanius et al., 1997). It has been
shown that genetic compatibility can have and influence on mate choice (Neff and
Pitcher, 2005). If potential parents tended to prefer mates with dissimilar alleles at the
highly variable MHC, it would be a powerful mechanism for avoiding inbreeding
depression. This alone would give a selective advantage to the maintenance of MHC
diversity, but of course, it would also augment immunocompetence. There are two ways
that MHC-dependant mating preferences could enhance parasite resistance. Firstly, the
MHC disassortative mating preferences of choosy parents would increase overall
population fitness because they would maximize the production of high-fitness
heterozygotes. Secondly, these mating preferences would be providing a means of
keeping up the co-evolutionary arms race with the parasites (reviewed in Bernatchez and
Landry, 2003). This scenario was coined as the moving target hypothesis by Penn and
Potts, in 1999 (reviewed in Bernatchez and Landry, 2003). It has been shown that MHC-
linked olfactory receptor genes are involved in discrimination and that MHC peptide
ligands are used as olfactory cues (Boehm and Zufall, 2006). For instance, this is the
case in three-spined sticklebacks (Gasterosteus aculeatus) (Milinski et al., 2005). In
salmon, mates significantly preferred to reproduce with fish with dissimilar alleles in
terms of overall amino acid composition. Whereas, rather than preferring males with
significantly dissimilar alleles, female stickleback preferred the odor of males with larger
numbers of alleles to that of males with fewer alleles. This suggests that the selective
mechanism favoring disassortative mating may differ between both species (reviewed in
Bernatchez and Landry, 2003). Despite this possible ambiguity, it is clear from this
evidence that the mechanisms of mate choice, as well as those of direct selection based
on resistance to pathogens, could play important roles in both the avoidance of inbreeding
and the maintenance of MHC diversity.

Evidence of selection at the MHC:

The neutrality theory indicates that nonsynonymous substitution mutations should be
selected against, as such mutations are deleterious to the function of most proteins. The
theory also states that synonymous mutations should accumulate over time, as they are
invisible to selection and thus are not weeded out over time. However, greater variability
is obviously a fundamental advantage for a protein which functions to recognize foreign
antigens. Thus, Darwinian selection might favor nonsynonymous substitutions,
enhancing the variability of the MHC binding sites for the purpose of improving
immunological competence. Indeed, significantly more nonsynonymous than
synonymous substitutions have been found in the functionally important antigen binding
regions of the MHC proteins where polymorphism was highest. The non-binding region,
however, has been shown to have a rate of nearly zero for both nonsynonymous and
synonymous substitutions (Froeschke and Sommer, 2005). Therefore, a protein with a
high proportion of nonsynonymous substitutions is a clear sign of positive selection and
is characteristic of a protein with antigen-presenting function (Schad, Sommer and
Ganzhorn, 2004).
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Another sign that can be used to infer the long-term effect of selection on the MHC
is trans-species polymorphism, which is the non-neutral retention of alleles across
species, causing discordance between alleles and species trees (Figueroa et al., 1988).
This makes sense if you consider that a speciation event can occur in a region where the
newly divided species are exposed to similar pathogens. Thus, despite being genetically
isolated, the two species would retain certain alleles of the MHC to maintain their
respective fitnesses. According to Coalescence theory, neutral polymorphism does not
persist for very long after a speciation event. However, Takahata and Nei demonstrated
in 1990 that polymorphism can persist under several models of balancing selection that
slows down the coalescence process, leading to long genealogical relationships among
alleles that transcend species (reviewed in Bernatchez and Landry, 2003).

Influences other than direct, gene-based selection:

Most vertebrate species show high levels of MHC diversity, but there are some
species that show little variation, such as the cheetah (Aconyx jubatus; Yuhki and
O’Brien, 1990) and the Fin whale (Balaenoptera physlaus; Trowsdale et al., 1989).
Again, this may merely show that such species are not under very much selection
pressure from pathogens, and so development of high MHC variability over time has not
been warranted, but that they may well be susceptible to novel pathogens. However, this
pattern could also be showing that behavioral and demographic factors may sometimes
play a larger role than balancing selection in some species (Miller and Lambert, 2004).
For instance, it is possible that there is little opportunity for MHC-dependant female
choice when other reproductive mechanisms are in effect, such as competition between
males for access to mates. This may explain why no MHC-dependant mating preference
was detected in Soay sheep even though there was an association between MHC variation
and juvenile resistance to pathogens (Paterson and Pemberton, 1997). Monogamous
mating systems may also contribute to low variability (Sommer et al., 2002). Thus,
social interactions may influence the intensity of balancing selection acting on MHC
polymorphism in natural populations (Hambuch and Lacey, 2002).

The overriding short-term influence of genetic drift:

However, for the most part, low variability in the MHC is associated with small
population size or recent population bottlenecks (Hedrick et al., 2000). Although
balancing selection seems to maintain MHC diversity over time, short term neutral
forces, causing genetic drift, sometimes override the effects of selection (Seddon and
Baverstock, 1999; Hedrick et al., 2001). The resulting short-term reduction of genetic
variability at the MHC is certainly evident following a dramatic bottleneck. Thus,
changes in MHC diversity can provide evidence of bottlenecks (Oosterhout, et al., 2006).
It is also clear that there is a relationship between balancing selection and effective
population size, as population size determines how vulnerable a population is to short-
term genetic drift. The generation and maintenance of MHC variation in New Zealand
robins appears mainly to be the result of balancing selection, although the short-term
forces determining MHC composition in their bottlenecked populations remain neutral
(Miller and Lambert, 2004). It should be re-emphasized that the example of the study,
conducted by Osterhout, et al. in 2006, showed the small Trinidadian guppy population
maintaining MHC variability comparable to that of a much larger population. This seems
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to show that genetic drift is merely a short-term factor and that balancing selection can
sometimes maintain high MHC variability even in small populations. However, this is
true only as long as the necessary selection pressure is present and only as long as the
process of balancing selection is allowed to perform over the long-term.

What is yet to be learned:

Although great strides have been made in our understanding of the MHC and the
forces influencing its diversity, there is obviously a great deal yet that must be
investigated. For instance, the amount of MHC diversity required to ensure long-term
population viability remains a fundamental question in the field of conservation genetics
(Miller and Lambert, 2004). Although there are known populations that appear to be
viable despite low MHC variability, it is generally thought that this is due to low
selection pressure resulting from isolated lifestyles and low exposure to pathogens. Such
populations may still be vulnerable to novel pathogens (Hambuch and Lacey, 2002).
Balancing selection has been revealed in humans and in murid model organisms
under controlled conditions for very few genetic systems, but the MHC system is one of
them. However, studies on free-ranging, wild populations are still somewhat lacking
(reviewed in Bernachez and Landry, 2003). It is only through the study of wild
populations that the practical, natural results of the influences of balancing selection and
genetic drift in populations can be studied. In addition, even among studies that
investigate wild populations, there is a strong taxanomic bias, favoring mammals more
than all other vertebrates combined. In order to generate a more complete picture of the
existing patterns of MHC diversity in natural populations, there will need to be better
taxanomic coverage in the future (reviewed in Bernatchez and Landry, 2003).

Conclusions:
It is clear from the available literature that the mechanisms of balancing selection, be
they based on resistance to pathogens or on disassortative mating, are the primary cause
of the generation and maintenance of genetic variation at the MHC in fish. Many factors
must be considered, such as population size, selection pressure from environmental
factors, and internal mating competition, in order to understand how the variability of the
MHC is being influenced. However, despite the power of balancing selection to promote
MHC variation over time, genetic drift due to short-term, neutral events, such as sudden
population bottlenecks, can override this process and reduce genetic variation.
Nevertheless, even a small population can show high MHC variability as long as the
necessary selection pressure is applied to it over many generations. As some species
seem to be viable with differing degrees of MHC variation, it is not clear how much
variation is generally required for long-term survival. As this may be an essential factor
for species conservation efforts, more research must be done, with an emphasis on studies
of wild populations across a wider range of taxa.
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Works Cited

Apanius V., Penn D., Slev P.R., Ruff L.R., Potts W.K. (1997) The nature of selection on
the major histocompatibility complex. Critical Reviews in Immunology, 17, 179–224.

Arkush, K.D., A.R. Giese, H.L. Mendonca, A.M. McBride, G.D. Marty, and P.W.
Hedrick. (2002) Resistance to three pathogens in the endangered winter-run Chinook
salmon (Oncorhynchus tshawytscha): effects of inbreeding and major
histocompatibility complex genotypes. Can. J. Fish. Aquat. Sci. 59, 966–975.

Bernatchez, L., and C. Landry. (2003) MHC studies in nonmodel vertebrates: what have
we learned about natural selection in 15 years? J. Evol. Biol. 16, 363–377.

Boehm, T. & Zufall, F. (2006) MHC peptides and the sensory evaluation of genotype.
Trends Neurosci, 29, 100–107.

Cable, J., and C. van Oosterhout. (2006) The role of innate and acquired resistance in two
natural populations of guppies (Poecilia reticulata) infected with the ectoparasite
yrodactylus turnbulli. Biol. J. Linn. Soc. in press.

Coltman D.W., Pilkington J.G., Smith J.A., Pemberton J.M. (1999) Parasite-mediated
selection against inbred Soay sheep in a free-living, island population. Evolution, 53,
1259–1267.

Figueroa, F., Gunther, E. & Klein, J. (1988) MHC polymorphism pre-dating speciation.
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Froeschke G. and Sommer S. (2005) MHC Class II DRB Variability and Parasite Load in
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Hambuch T.M., Lacey E.A. (2002) Enhanced selection for MHC diversity in social tuco-
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Hedrick P.W., Gutierrez-Espeleta G.A., Lee R.N. (2001a) Founder effect in an island
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Hedrick, P.W., and K. J. Kim. (2000) Genetics of complex polymorphisms: parasites and
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Jeffery, K.J., and C.R. Bangham. (2000) Review: do infectious diseases drive MHC
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by MHC peptide ligands. Proc. Natl Acad. Sci. U.S.A., 102, 4414–4418.

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Meyer-Lucht, Y., and S., Sommer (2005) MHC diversity and the association to nematode
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Mohammed, R.S., Persad, N., and Cable, J. (2006) Balancing selection, random
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