Chemical Communication: The Language Of Pheromones

PREFACE

About twenty years ago, some of my colleagues in another laboratory at The Rockefeller University were investigating the Syrian golden hamster. They were using the hamster’s sex attractant to explore biological and behavioral questions, and they recognized that knowledge of the chemical compounds responsible for the hamster attractant could facilitate their research. No one had ever examined this pheromone chemically, and they soon found that doing so would require a serious chemical investigation. At this point they encouraged my research group to take up the problem.

Now, my research group spent all its time exploring details of how chemical reactions work. Although we were very familiar with handling chemical compounds, we had never dealt with chemical signals or hamsters. Working with chemical reactions seemed quite different to me from investigating small rodents. Nonetheless, the idea intrigued me, because at that time no one knew much about the chemistry of mammalian pheromones. I was interested and agreed to plunge in.

In making this decision, I understood that our investigation would move much faster if the experiments were in the hands of someone already experienced with pheromones. I began looking for an investigator with an appropriate background and very soon found just the right person. Dr. Alan G. Singer had just completed his doctoral research, isolating and identifying a mammalian pheromone, and the experience had given him ideal preparation for the hamster problem. I persuaded him to join us at Rockefeller as a postdoctoral fellow and to devote his full attention to the hamster attractant. Over the next two years he first determined the identity of the major hamster sex attractant and then turned his attention, with similar success, to another signal that is a powerful aphrodisiac for the male hamster.

Our accomplishments rightly reflect Singer’s ability and effort, but they also owe a great debt to an unusually effective interdisciplinary collaboration with two groups of biologists. Because no one is equipped to work equally well in all the areas that contribute to the study of pheromones, cooperative research has had an important role in understanding chemical communication.

The scientific study of pheromones combines elements of several disciplines, but many of the results can be understood and discussed without much technical language. This volume brings together information about chemical signals that carry a wide variety of messages in species from microbes to man. No particular knowledge of animal behavior, sensory physiology, or organic chemistry is required to follow the discussion, although enough technical detail and reference material are provided to permit the prepared reader to explore further. I have presented the chemical structures of the compounds responsible for many of the pheromones discussed, even though we do not yet understand much about why one structure and not another serves to carry a specific chemical message. I appreciate that most people (including scientists in many other fields) do not share the organic chemist’s strong esthetic sense for molecular structures, but for me the story of pheromones would have been incomplete without their chemical identities. Of course I hope that you will agree, but a reader less enthusiastic about chemistry should be assured that many behavioral scientists pursue significant research into chemical communication without giving more than a passing thought to chemical structure.

Discussions of pheromones can be organized around either messages or organisms. I suspect that for most people biological species are more familiar than the signals these species use, and for this reason I have organized the discussion around organisms. This approach also emphasizes the grand sweep of diverse living creatures that make use of chemical communication. I have started with more simply organized species and moved toward more complex ones. Some scientific problems associated with chemical signals grow more difficult as the organisms using the signals become more complex, and moving toward increasing complexity provides a gradual approach to these difficulties.

I began writing this book wondering whether it would find an audience. The subject attracted me, but had enough people heard of pheromones to be interested in reading about them? I was recently reassured on this point. When two New York socialites kissed and made up immediately after a well publicized spat, a local gossip columnist explained confidently that it was a chemical thing involving pheromones. This disclosure came to my attention, and I was delighted to learn that chemical signals had achieved such a prominent place in the world. This should certainly be an appropriate time to say something serious about them.

William C. Agosta

New York, March 1992

1

MOLECULAR MESSENGERS

e9781466813939_img_9473.gif A female silkworm moth, perched on a cocoon, emits a chemical signal that attracts a mate. So small is the amount of chemical produced that to isolate and identify the substance scientists needed twenty years and over half a million female moths.

Human beings gather information about the outside world largely through sight and sound. These are our best-developed senses, and when we want to know what is going on we look and listen. For many other living creatures, the world is a much different place: for them chemical signals are the primary source of information. Chemical compounds from other organisms or events in the environment provide their basic knowledge of the world. Even many creatures that do have other well-developed senses find chemical signals to be indispensable Species as diverse as water molds and elephants depend on the ability to sense chemicals in the world around them for survival. Even simple bacteria respond efficiently to certain chemicals, moving toward foodstuffs and away from toxic compounds. Humans must have developed an early appreciation of chemical signals. The attraction of a male dog to a female, for example, could have provided important clues thousands of years ago. Even earlier, hunters must have learned about the significance of scent to both prey and predator. However, our organized, scientific understanding of chemical signals is a much more recent development. This understanding began in the nineteenth century with the experiments of naturalists who were intrigued by the ability of female moths and butterflies to attract the male. For decades these investigators documented attraction for various species, often over distances of several kilometers. Through clever experiments they demonstrated that functional antennae were critical for the male to locate the female: males could no longer find females when their antennae were covered with lacquer or totally removed.

Dogs commonly sniff at one another, as depicted in this lithograph by Pierre Bonnard. They are a thousand to a million times more sensitive to various odors than we are, and recognize each other by smell.

These observations were fascinating, but scientists achieved a satisfactory interpretation of them only slowly. Initial explanations invoked some sort of radiation from the female, and one idea was that the male’s antennae were appropriately tuned to receive the wavelength broadcast by the females of his species. As late as 1950, investigators found that infrared radiation emanated from the region of the thorax of females of two species of moth. In their report of this discovery, these scientists wondered if this radiation might be the signal emitted by the female to lure her mate.

On a more rewarding track, as early as the 1930s there were reports of the extraction of attractivity from female moths. There was also evidence that attractive substances were present in the air over cages containing females. Experiments eventually showed convincingly that attractivity must be due to volatile chemical substances. At the end of the 1930s this conclusion was firmly enough established that the German chemist and Nobel laureate Adolph Butenandt became interested in undertaking the isolation and identification of such a sex attractant. After some twenty years this research led to the identification of bombykol, the sex attractant of the female silkworm moth Bombyx mori. This investigation holds an important place in our understanding of pheromones, and we shall return to it later.

About half the 15,000 to 20,000 hairs on the feathery antennae of a male silkworm moth are specialized for detection of the sex attractant released by the female.

At about the time that Butenandt completed the work on bombykol, Peter Karlson and Martin Luscher coined the word pheromone to describe a chemical signal transmitted between members of the same species. (The word was created from two Greek words, pherein, to transfer, and hormon, to excite.) They wished to distinguish pheromones from other kinds of chemical signals that convey information to organisms. Some of these other kinds of signals come from whole systems such as rivers or forests, as when the odor of its native stream guides a salmon returning after years in the distant ocean. Other chemical signals pass from members of one species to another: dying trees release substances detected by bark beetles that then attack the trees; skunks defend themselves against other animals with an unforgettably malodorous spray. None of these examples concerns a message sent and received within a single species, and so none of these signals is a pheromone. There is also another kind of chemical signal that is not a pheromone, even though it does concern a single species. This is a signal that operates within a single individual, delivering a message from one part of the organism to another. These signals are known as hormones, and they will turn up again later in our explorations.

While Butenandt was setting out to isolate bombykol, the famous behaviorist Karl von Frisch was making one of the classic discoveries of chemical communication. An Austrian zoologist most widely known for his work with honey bees, von Frisch (1886—1982) shared in the Nobel Prize for Physiology or Medicine in 1973. His contribution to the study of pheromones emerged from his work with the European minnow Phoxinus phoxinus, and it came about through two casual observations. The first concerned a minnow that von Frisch had marked with an incision near its tail. When he returned this individual to the school, the other minnows became frightened and some of them retreated. The second observation followed von Frisch’s finding a minnow stuck under the rim of a feeding tube, struggling to free itself. He released the fish, and when it swam away toward the group, they all fled in fright. To von Frisch these episodes were curious enough to warrant detailed investigation, and his carefully documented results set the direction of much later research into chemical communication in fishes.

A school of young European minnows.

Von Frisch called the agent responsible for this fright reaction the alarm substance. Subsequent work has shown that most members of this group of fishes (the ostariophysians) show a fright reaction when an injured ostariophysian is introduced into the water nearby; the nature of the reaction itself varies with species, sex, and location of the reacting fishes. They may swim in a tighter school, leap at the surface, swim away and hide, or sink to the bottom and become very still. In some instances they will avoid the area of their disturbing encounter for some time thereafter. In at least one species the fright reaction changes over the life of the fish: young creek chub (Semotilus atromaculatus) dart away and hide, but adults drop quietly to the bottom and remain motionless.

This sudden and remarkable reaction is under the control of a pheromone in the skin of the fish, where it is carried in large alarm-system cells. These cells are fragile, and upon injury they rupture, discharging the pheromone into the water. Simply scaring a fish does not affect its alarm-system cells, but even minor mechanical damage to the skin will disrupt them, and damage to only a small area of the skin of a single fish is capable of causing fright in an entire school.

The alarm substance and bombykol are just two of thousands of pheromones. All sorts of organisms use chemical signals to convey a rich variety of messages. The alarm substance signals danger—get out of here!, and bombykol is the female moth’s message come to me. Other pheromones bear messages such as the queen is in the hive and all is well, produce more sex hormone, we are under attack! and I am pregnant. Messages may elicit a specific behavioral response in the recipient, such as swimming toward the source of the signal or attacking an enemy. Alternatively, the message may cause a physiological change in the recipient, perhaps altering the timing of the sexual cycle or inducing puberty. For some species we are aware of only one pheromone and a single message. Other species, such as ants and honey bees, use many different chemical signals to coordinate the activities of their complex communities.

The large alarm-system cells of the European minnow are located in the outer layer of the skin, where they can be ruptured easily upon injury. The mucus cells discharge the slimy substance that forms the protective outer coating of fish.

As we consider the significance of pheromones in the living world, it is important to keep in mind that human beings make relatively minor use of the ability to sense chemicals in the environment. Our primary means of detecting chemicals is through olfaction, the sense of smell. The sensitive discrimination of professional perfumers and wine tasters demonstrates that considerable refinement of the human olfactory ability is possible, but we are much less sensitive to chemicals than many other species. It is quite normal for some people to be a thousand-fold more sensitive than others to specific chemicals, evidence of how variable the importance of olfaction is to humans. Without a sense of smell, we would certainly find the world a much poorer place, but for us olfaction is not critical to survival or even to a reasonable existence. People who lack a sense of smell are said to suffer from anosmia. This condition accompanies several serious maladies, but there are many anosmic people who show no other defect and who live quite normal lives. For human beings, anosmia is not even remotely comparable to the losses entailed in blindness or deafness. In contrast, chemical communication plays an indispensable role in the lives of the creatures that we shall discuss in the pages that follow.

When disturbed, honey bees release an alarm pheromone and fan their wings to disperse the signal to their nest mates.

Pheromones as Chemical Compounds

Pheromones are specific chemical compounds or mixtures of compounds. This means that they are real objects that are endowed with definite physical and chemical properties. Once released, a pheromone has a physical existence quite apart from the organism that produced it. A volatile chemical signal may be carried on the breeze or in a current of water, to deliver its message at a later time and in a place remote from its source. Such a signal could be effective at attracting faraway mates or helpers. Chemical signals may also be persistent. Some of them can be deposited on a bush or on the ground, to be detected where they were left after the sender has departed. If such a signal is chemically stable and not too volatile, it may remain in place, and active for days. Stability and nonvolatility are good properties for a signal used, for example, to delineate the boundaries of a territory or to mark a food source. Conversely, the signal may be chemically unstable and destined to provide its message only briefly. This might be useful in a pheromone employed to give an immediate and short-term warning of danger.

Pheromones that travel through the air need to be volatile, ones that are released in the ocean must be stable in water, and ones that must remain in one place for an extended period should be persistent. Virtually all the compounds that will appear in the pages to follow are organic molecules. Molecules are groups of atoms assembled into a specific three-dimensional arrangement or structure, and organic simply means that these molecules contain carbon atoms. Properties such as volatility, persistence, and stability have their basis in molecular structure. The smaller a molecule of a given kind of is, the more volatile the compound will be. Two simple molecules illustrate the effect of size: pentane has 5 carbon atoms and pentadecane has 15, but otherwise they look much alike; yet the small size of the pentane molecule leads to high volatility. A small amount of pentane poured into a saucer will evaporate in a minute or so, whereas the much larger pentadecane molecules form an oily, persistent substance. A small amount does not evaporate noticeably in hours.

Like many other cats, cheetahs (Acinonyx jubatus) send a spurt of urine backward to mark trees and other conspicuous objects. This probably permits small groups of cheetahs to avoid each other while hunting in the same