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Determining factors for the growth and colour of the mane in the African Lion (Panthera leo)
D.A. Conradie# (29033196)
Department of Animal and Wildlife Sciences, University of Pretoria, Private bag X20 Hatfield, Pretoria 0028, South Africa

___________________________________________________________________________________ Abstract
Lions are the only species in the Felidae family that have evolved a mane. The lion mane, which is only found in males, is one of the most distinguishable features in the lion that separates it from the other individuals in the Panthera genus. The mane is said to have evolved 320 000 to 190 000 years ago (Yamaguchi et al, 2006), after the lion diverged from the common ancestor it shares with the other members of the Panthera genus; namely the leopard (Panthera pardus), jaguar (Panthera onca), tiger (Panthera tigris), clouded leopard (Neofelis nebulosa) and the snow leopard (Unica unica). The mane is thought to have evolved to serve as an indicator of health, maturity and testosterone levels. It is used for protection during battles and to intimidate other males. Lastly, it is thought to attract females as it portrays the males masculinity. The mane serves as a social benefit with ecological costs (Patterson et al, 2006) as the mane has a huge cost in the form of extra heat. Climate therefore also plays a large and important role in mane growth, with hotter areas tending to produce lions with smaller or no manes. The colour of the mane is said to be affected by maturity becoming darker and longer with age. A hormone effect is also evident where individuals with higher testosterone levels have a darker mane. Nutrition plays an important role in both mane growth and colour healthy, well fed males tend to be stronger with a larger, denser and darker mane. Although much research has been done by several different authors to try and explain the reason for the manes evolution, and subsequently its function today; there is as of yet no concrete evidence for its evolutionary purpose. The exact mechanism behind its growth and development is also not yet fully understood, and largely speculative. __________________________________________________________________________________________ Keywords: evolution, protection, intimidation, attraction, temperature, climate, hormone, testosterone, maturity, health, nutrition
#Corresponding author: donne@woodlandsnet.co.za

Introduction
Lions are part of the Felidae family and the only species in the Panthera genus that have evolved groupliving behaviour and the characteristic mane evident in males. The mane is a sexually selected characteristic and the only real example of sexual dimorphism in the Felidae and Carnivora (Patterson et al., 2006). As the lion is the only species under the Panthera genus to have evolved a mane, it was suggested that the mane evolved after lions diverged from their common ancestor that they share with the other species in the Panthera genus (Yamaguchi et al., 2006). There is, as of yet, no consensus on the true function of the lions mane. It is proposed to have a role in intimidation, attraction and protection. Lions are generally large animals, with the average male weighing between 150 and 260 kg (Kayes & Patterson, 2002). A large, thick mane will make a male lion look even bigger and more intimidating, thus improves his chances of avoiding conflict with rival males. Fierce competition exists between male coalitions as all males try to fight their way into a pride (Iwago, 1995). Fights between rival males can be deadly; this further increases the advantage of having a large mane. The mane also acts as an advertisement to females. In a study by West and Packer (2002), it was shown that females are more inclined to approach a dark-maned male compared to a light or blonde maned male. Lastly, it is suggested that the mane has evolved and functions as a form of protection as it covers vital areas such as the skull, neck and chest. Charles

Darwin (1871) was one of the first to classify the mane as a form of protection. However, there is no concrete evidence that shows that the mane has a protective function, as portrayed in a study done by West et al. (2005). The variation in mane colour is another aspect of the lions mane that is often discussed. The influence of hormones, specifically the androgen testosterone, is the most accepted reason for the darkening that occurs as a male ages (Randall, 2008). Other suggested influences include nutrition and health as these factors determines the availability of nutrients to allow for melanisation of the hair follicles. Although temperature and climate has also often been suggested, a study done by Patterson et al. (2006) on the effect of climate on mane variation in zoo lions showed no correlation between temperature and mane colour.

Evolution of the mane and its function

The mane evolved between 320 000 and 190 000 years ago during the Pleistocene era in the east of Africa (Yamaguchi et al., 2006). During the late Pleistocene era, lions were found across the globe and not concentrated in Africa like they are today; with fossils of lion-like felids found in Africa, Europe, Asia, North America and the northern parts of South America. The lion is the only species in the Felidae family that has evolved a mane, therefore it is accepted that the lions mane evolved after the lion diverged from the common ancestor that it shares with the other members of the Panthera genus (Yamaguchi et al., 2006). The mane has been described as a social benefit with ecological costs (Patterson et al., 2006). The evolutionary purpose of the mane is a topic that is frequently discussed. Evolution can be defined as a process of gradual change in a group of individuals over successive generations in which favourable traits are selected for, causing a change in the frequency of specific traits. As evolution only favours favourable traits, the presence of a mane must have served as a major advantage to the male lion, especially when considering its costs. Firstly, the mane has a cost on thermoregulation as the mane serves as a heat insulator. The insulation properties of the mane was confirmed in a study done by West & Packer (2002) in which they measured the surface temperature of maned and maneless males through infrared thermography and compared it to that of female lions to determine the added heat cost of the mane. Secondly, the mane has an added cost to maintenance due to the extra need for nutrients and energy for the production of extra long hair follicles; this will however not be a problem in areas where food is abundant. Lastly, the mane makes the male more conspicuous to prey and makes it more difficult to move through thick bush (Kayes & Patterson, 2002), which are both factors that can hamper the lions ability to hunt. This may in turn have had an influence in the development of group-living behaviour that is primarily seen in lions; where females, who are less hampered by a bulky mane, do most of the hunting. However, group-living has been documented to not be a function of hunting success, but rather of the competitive advantage in intergroup competition (Mosser & Packer, 2009). Although there is no concrete evidence for a single function that aided in the evolution of the mane, there are some suggested functions that have been widely researched and discussed; these include protection, advertisement and intimidation. Protection The most common suggested function of the mane is for protection. The mane covers the forehead, neck, shoulder and chest, which are all relatively vital areas. This strengthens the suggestion that the mane acts as a form of protection. It has further been hypothesised that darker, longer hairs are stronger than shorter, blonde hairs and provides more protection. Hair strength is determined by examining the amount of breakage at the distal end of the follicle and is indicated by jagged ends as opposed to flat ends (Braida et al., 1994). The theory of protection can however be argued. To begin with, the lion is the only felid to have evolved a mane, even though all members of the Felidae family have the same weapons and are similarly armed (West and Packer, 2002). In a study done by West et al. (2005), it was also shown that longer hairs were more prone to breakage as indicated by jagged ends, although darker hairs were shown to be less broken as compared to blonde hairs. The study further investigated the proposed protection hypothesis by evaluating the amount of attacks to the areas covered by the mane, the incidence of wounds in the areas covered by the mane in females and subadult males, the rate of mortality due to inflicted wounds in the mane area and evaluated whether the rate of

mane development at the different initiation points reflects the association between the frequency and mortality rate of wounds in the respected areas. In the study by West et al. (2005) it was found that adult males were wounded significantly less on the mane area in comparison to females and sub-adult males and suggested that the mane area may be avoided by attackers. It was noted, however, that their results may have been biased due to the mane obscuring some wounds. As for mortality rates associated with wounds to the mane area; the study showed that wounds to the mane area was not more dangerous or fatal compared to those on other areas of the body. It was also added that wounds on the forehead and face are more difficult to lick and clean, therefore may result in a higher than necessary mortality due to the inability to clean the wounds. Lastly, with regards to the rate of mane development; if protection was the main function, the mane should develop earliest and fastest on the forehead. However, the study revealed that mane development began on the chest and neck and that it only later started developing on the forehead. With regard to the colour of the mane playing a role in protection, the study by West et al. (2005) showed that the mane begins darkening at the neck and shoulder and that chest mane hair was the darkest. It has also been shown that darker hairs are thicker than blonde hairs, thus provides more protection. Intimidation Between-male conflict can be deadly. Male lions compete not only for access into prides, but also within male coalitions for mating opportunities. Due to the brutality of conflict, a male lion would want to avoid challenging another male that is a lot stronger than he is. This is where the mane may play an important role. The mane mainly grows and darkens with age, but the condition of the mane has also been suggested to be an indication of health status and testosterone levels. As testosterone in males is linked to aggression (Book, 2001), and mane colour is said to be influenced by testosterone, a darker maned lion would be portrayed as more aggressive. This was evident in a study done by West & Packer (2002) where their study showed that males were more hesitant to approach a longer, darker maned lion. The size of the mane will also play a role. It is commonly know that appearing larger makes an individual seem more intimidating. This will have the same effect in lions, where larger maned males seem visibly larger and are therefore perceived to be stronger and more dominant. Attraction It has been shown that lionesses are more attracted to darker maned lions than lighter maned lions (West & Packer, 2002). This could be indicative of the visual role the mane plays in attracting females. It has been suggested that males with darker manes have offspring that survive longer, thus a favourable choice to a lioness to ensure the survival of her cubs. When a new male takes over a pride, he kills all the defeated males cubs to ensure that the lionesses return to heat and that only his genes are be passed. It then makes sense for a lioness to choose a darker maned male, as his chances of being overthrown by a rival male would be less due to his higher testosterone levels, visual intimidation and perceived aggression. Mane length was shown not to play a role in attracting females (West & Packer, 2002).

Factors affecting mane growth

There are significant differences in mane length for different areas of the body; the forehead being the shortest, chest mane being significantly longer than neck mane, while neck and shoulder mane is similar in length (West et al., 2005). The mane develops with age, starting at the age of 12 months and is usually fully developed by the age of 4. There are, however, several factors that have been suggested to have a role in mane development; these include season and climate, hormones, maturity, health and nutrition. Season and climate Due to the high thermal cost of a larger mane, it can be considered that mane length is inversely correlated to environmental temperature. This has been shown in several studies where lions at higher altitudes develop larger manes, while manes are shorter in hot climates, for example the Tsavo lions.

In a study done by Patterson et al. (2006), mane length was compared among zoo lions across America. Since nutrition, health, social structure and hormone levels were relatively constant, environmental effects could play a larger role in mane development, while genetic effects, which are lowly heritable, could be related to the hormone levels which govern mane growth. It was shown that climate does have a large effect on mane growth as it caused up to half of the variation in mane length. An interesting discovery was that cold weather, below the lions thermoneutral zone of 9 to 33C (McNab, 2000), was more closely correlated to mane length than hot temperatures and heat stress. It was suggested, however, that measuring mane length in spring rather than autumn depicted the effect of the winter coldness and not that of the heat stress experienced in the summer. However, there is contradicting information on the seasonal variation of the mane with some studies supporting it (Patterson et al., 2006; West and Packer, 2002) while others found no supporting evidence (Gnoske et al., 2006). Hormones Hormones play an essential, intricate role in hair growth as it coordinates growth with age and stage of development or environmental alterations (Randall, 2008). As I have already mentioned; the mane is a sexually dimorphic trait. Sexually dimorphic traits in vertebrates develop secondarily to the reproductive organs, namely the ovaries and testes, under sex-specific hormone influences (Williams & Carrol, 2009). It can therefore be accepted that male-limited traits, like the mane, is under the influence of testosterone or testosterone derivatives, which in turn would be under sex-limited gene regulation. Androgens are the main regulators of mane growth in lions and act locally on specific receptors in the skin (Messenger, 1993). As testosterone is a steroid hormone, it can diffuse directly through the hydrophobic lipid bilayer of the cell membrane. Testosterone will follow one of two routes; it either moves directly into the nucleus where it binds to a receptor, or it is first metabolised into a more potent form, 5-dihydrotestosterone, which binds to the receptor. The hormone-receptor complex then binds to the DNA binding site and initiates gene expression through transcription (Randall, 2008), causing hair growth. It has also been showed that castrating adult males may cause them to lose their mane, and prevents mane growth in subadult males (Guggisberg, 1961). This is another indicator of the importance of testosterone in mane growth as castration involves the removal of the testes and subsequently the major testosterone producing organ. Maturity By looking at the mane types that were used in the study by Gnoske et al. (2006) to classify and score mane development, one can clearly see the development progress of the mane. Growth of the mane starts at roughly 12 months of age with the growth of sideburns and bilateral chest hair at the shoulder joints and a dorsal neck crest on the scruff of the neck. Mane development then spreads to the neck and chest, growing longer and filling out as the young male ages. By the age of 4, the mane covers the whole head and neck and is considered fully grown. Mane growth will continue over the shoulders and down the chest as the male grows older and his mane grows longer and bigger. Health and nutrition During a study done on zoo lion mane variation (Patterson et al., 2006), it was shown that zoo lions have significantly larger manes than free-living wild lions. This can be indicative of the differences in nutrition and health. Zoo lions are fed more often and are fed a more balanced, nutritious diet. They are also monitored more closely and are treated for diseases and infections. Hair growth, and especially the mane, has a high metabolic requirement (Randall, 2008). Hair is composed mainly of keratin which forms the intermediate filaments in the hair follicle. Being a protein, the production of keratin and therefore hair growth as a whole, will be negatively influenced when lions are not eating. It would therefore make sense that males not receiving enough protein would have impaired hair growth. Unfortunately, health is a larger issue for males than it is for females. A sick or injured male cant defend his pride and will be overthrown by a stronger, healthier rival male. Hunting is easier when in a pride as

the lionesses go out and hunt together. A defeated male will have to hunt for his own food, a task even more challenging if sick or injured. The extra activity needed for hunting further increase his energy demands and decrease the protein available for hair growth.

Factors affecting mane colour

As a male ages, so his mane becomes darker, changing from tan, or blonde, to almost black. Usually only the neck, chest and shoulder hair darkens, with the hair on the head remaining the usual tan colour. Several factors are considered to influence the colour of the mane; these include the climate, hormones, maturity and genetics. Climate and environment It is known that animals living in hot, desert environments have usually adapted a lighter pigmentation compared to darker pigmented animals living in cooler, more humid areas, like forests; which is in line with Glogers rule (Cowles, 1957). This is due to the heat attracting properties of the colour black that will add an extra heat cost to the animal. Dark hair follicles were also shown to be thicker than lighter hair follicles, hampering heat exchange and thermoregulation in hot climates (West & Packer, 2002). A darker mane would therefore attract more heat than a lighter one, increasing the already high heat cost of a mane. Another climatic effect can be reflected in the environment. Hot savannahs are usually dull in colour. Males with lighter, tan coloured manes would therefore be camouflaged in their environment. A black mane goes against this, making the lion more conspicuous to its prey as it stand out from its environment. However, it has been shown that climate and environment does not play a large role in mane colour variation (Patterson et al., 2006) as male lions from all regions and all climates have dark manes. More research needs to be done the climatic effect on mane colour to establish its true effect. Hormones Melanisation is the process responsible for the production of colour pigments from tyrosine. Melanin is divided into different categories depending on its colour; eumelanin produces brown to black pigmentation while pheomelanin produces yellow to reddish-brown pigments. The tan colour usually seen is therefore produced by pheomelanin while black manes are produced by eumelanin. The formation of eumelanin and pheomelanin is regulated by the amounts and presence of glutathione and cysteine which modifies the melanogenic pathway; low cysteine and high glutathionine produce eumelanin while high cysteine and low glutathionine produces pheomelanin (Slominski et al., 2004). Testosterone plays an important role in the initiation of the melanisation process as it targets androgen receptors in the hair follicle. It is metabolised in melanocytes to 5-dihydroxytestosterone which initiates the transcription of 5-reductase, which then plays a role in melanisation. Therefore, males with a higher testosterone level will have increased melanin production and produce a darker mane under the right circumstances. Maturity The mane only starts to darken after a male reaches sexual maturity, and then darkens as he ages. Sexual maturity is reached in males at roughly the age of 3 (Brown et al., 1991), which is when the mane is almost fully developed. Sexual maturity is accompanied by an increase in the hormone testosterone as the testes become functional. We can then again refer to the process of testosterone in production of eumelanin and the darkening of the mane. Genetics DNA and the genes it holds are the centre point of all biological processes in the body as these regulate the expression of proteins which act as regulators for almost every process in the body. Mutations or alternative forms of a gene, called an allele, can change the expression and regulation of proteins and biological processes.

It is often seen that different lion populations have males with darker coloured manes than others. Males with lighter mane colour may have differences in their DNA compared to a male with darker manes. These genes may then be passed on to its offspring, resulting in those males also having a lighter mane. A genome study was done by Dubach et al. (2005) that identified 6 genome haplotypes that could be used to identify lions from different regions in Africa; while 2 major geographic clades, a south-western and eastern Africa clade, were also identified. Unfortunately, this study only looked for differences in the genome so as to separate populations for phylogenetic purposes, and did not associate genotypic difference to phenotypic variations. Had they associated genotypic differences to phenotypic variation, it might have shed more light on the role that the genome plays, not only on mane colour, but also in mane growth as a whole.

Exceptions
There are two well known exceptions to the normal growth and colour pattern of the mane. The Tsavo lions of Kenya have a reduced mane growth and are often termed maneless, while white lions, on the other hand, haven abnormal white pigmentation in not only their mane, but their entire coat. The Tsavo lions Numerous studies have been done on the uniquely maned lions of the Tsavo region in Kenya. It has been widely accepted that these males, although fully mature, lack a mane. This has lead to an increased uncertainty on the true function and evolution of the mane as these lions go against the norm. A thorough study on the Tsavo lions was done by Gnoske et al. (2006) in which they compared the variation between the Tsavo lion populations in Kenya to a normal lion population in the Kruger National Park in South Africa in terms of mane morphology and environmental conditions. The study showed that not all Tsavo lions were maneless (only 7 out of the 55 lions observed). What they did find was that the onset of mane development was delayed; manes only started to develop between the ages of 3.5 and 4 years, while in normal populations the mane would be almost fully formed by then. Secondly, it was shown that mane growth was slower and more limited in the Tsavo population in comparison to normal populations, where manes were only fully developed after the age of 7. Even when fully grown, the Tsavo lions manes werent as large or well developed as compared to those in normal populations. Another interesting find was that actively breeding males had smaller, lighter manes than would be found normally. This indicates that there is a dissociation between the primary and secondary sexual characteristics; where in normal populations, the age of mating is usually correlated to mane size. This also proves that mane size and fertility are not linked as there is no record of a diminishing lion population, even though breeding males have smaller, less developed manes. The lack of mane growth may be as a result of the extreme environmental conditions. The Tsavo region is located below an elevation of 800m with high humidity levels and high minimum temperatures. It can therefore be suggested that the more extreme climate had caused a change in the rate and degree of mane development, but did not compromise their reproductive ability (Gnoske et al., 2006) White lions The white lion has a white to cream colouration as opposed to the normal tawny phenotype. The white coat colour is due to a recessive autosomal gene that causes a lack of pigmentation in the fur. Although they have a loss of pigment in their coat, they still have pigmented eyes and can therefore not be classified as an albino, which dictates that all pigment is lost. White lions are rather classified as leucistic. Leucism is a form of incomplete, partial albinism (Forrest & Naveen, 2002) characterised by a reduced pigmentation, but still allows for the presence of pigmentation in the eyes, eye lids, nose and lips. In feline coat genetics, the dominant C allele codes for the tyrosinase enzyme which is necessary for the production of pigment; while the recessive or null-type allele results in an albino. A dominant, white masking gene (W) is responsible for a white coat colouration as it prevents the formation of melanoblasts in the skin and the subsequent expression of colour. However, it has been suggested that a recessive gene mutation, known as the Chinchilla gene, rather than a dominant white allele, is the reason for the white coat colour (Robinson & De

Vos, 1982) seen in the white lion. It is proposed that the Chinchilla mutant prevents the production of phaeomelanin and degrades the quality of eumelanin, thus producing a pale colouration. A similar mechanism may be the cause for the white colouration seen in the white tigers.

Conclusion
The true evolutionary purpose of the mane has not yet been pin-pointed, although several possible explanations have been suggested. The role of sexual selection, however, has played a definite role in its development as only males have a mane, while lionesses, who were exposed to the same environmental conditions, do not. Even though lionesses are equipped with the same weapons and armour during conflict, it can be argued that there is less between- and within-pride conflict among lionesses, although they are more vulnerable during hunting than males. A lot of research has been done to try and determine not only the evolutionary purpose, but also the mechanism by which the mane grows and develops. Mane length and colour is not regulated by a single factor, but rather by several factors including climate, hormones, maturity, health and nutrition. All of these are interlinked and have an effect on one another, making their individual effects harder to determine in free-living populations. Unfortunately, most of the research that has been done is not conclusive and more intense and accurate research will be needed. More specifically, research is needed on the individual effects of climate and testosterone levels on mane growth and development, as well as possible genome differences in natural, freeliving populations. This may, in turn, shed some light on the true reason for the manes evolution.

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