A New Key to the Story of How the Sexes Have Come to Be

In the beginning, we are all male and female… More specifically, we are all in between male and female. So what makes our embryonic selves choose and follow a developmental path to becoming the sex that we are today? New research has dramatically changed our understanding of this process.

During early embryonic development, all mammals develop a single pair of gonads that are neither testes nor ovaries, but have the potential to become either. Likewise, the external genitalia at this early stage has the potential to become either a penis and scrotum or a clitoris, vagina, and labia. Two pairs of ducts develop to connect the gonads to the undifferentiated external genitalia: One set of ducts, the Wolffian ducts, would become the epididymis, vas deferens, and seminal vesicles if this animal becomes male. The other set of ducts, the Müllerian ducts, would become the oviducts, uterus and innermost part of the vagina if this animal becomes female. So what determines if a given animal will develop male or female reproductive anatomy?

Early in development, mammalian embryos have one set of gonads that has the potential to become either testes or ovaries (here labeled as "bipotential gonad"). These gonads are connected the the developing external genitalia by two sets of tubes: The Wolffian ducts become the reproductive tracts in males and the Müllerian ducts become the reproductive tracts in females. The ducts that do not become reproductive tracts typically disintegrate. However, XX female embryos that lack the COUP-TFII protein do not dismantle their male-like reproductive tracts. Figure from Swain, 2017.

The sex of a mammal is determined by the combination of sex chromosomes it has. If the mammal has two X chromosomes, it will likely become female, and if it has an X chromosome and a Y chromosome, it will likely become male. The story physiologists have been telling for decades is that there is a single gene located on the Y chromosome, called the SRY gene, that single-handedly makes an embryo become a male. When expressed, the SRY gene produces a protein, called testes-determining factor, which interacts with the cells of the undifferentiated gonads to turn them into testicular cells. These newly formed testicular cells produce two key hormones: testosterone, which causes the Wolffian ducts to become the epididymis, vas deferens, and seminal vesicles, and anti-Müllerian hormone (AMH), which causes the Müllerian ducts to degenerate. In other words, if an animal has a Y chromosome, it will typically have an SRY gene that will trigger the sequence of events that causes the animal to develop into a male. If the animal does not have the Y chromosome, it will typically become female. However, it is not just the lack of a Y chromosome that can make a female; Any disruption of this pathway (such as an SRY gene that is not expressed, or the lack of testicular hormones) typically causes the animal to develop into a female. For this reason, females in mammals have been called the default sex. The scientific understanding since the 1950s has been that, in mammals, the development of a male reproductive system is an active process and the development of a female reproductive system is a passive process. However, a new study reveals that the process of becoming female mammal is not as passive as we have thought.

Fei Zhao, Humphrey Yao and their research team at the National Institute of Environmental Health Sciences and Baylor College of Medicine discovered a critical role for a specific protein, called COUP-TFII, in the active process of becoming a mammalian female. The research team examined female mouse embryos (which lack a Y chromosome, and hence an SRY gene) that had been genetically modified to lack a particular protein called the COUP-TFII protein. They compared these genetically modified XX embryos to genetically typical XX mouse embryos. When the unmodified XX embryos had developed to have only Müllerian ducts (the “typical” female reproductive pathway), the XX embryos without COUP-TFII protein retained both Müllerian and Wolffian ducts! Unfortunately, these XX mice that lacked the COUP-TFII protein died shortly after birth, so it was difficult to tell if this developmental process would have continued. The research team cultured reproductive organs of XX mice with and without COUP-TFII protein and found that this developmental trajectory likely would have continued after birth.

Images A and B show the reproductive tract from the side (A) and as a cross-section (B) in a "typical" XX female mouse embryo. Images D and E show that XX females that lack the COUP-TFII protein retain both Müllerian (pink arrows) and Wolffian (blue arrows) ducts. Figure from Zhao et al., 2017.

We know that testosterone helps promote the development of Wolffian ducts in XY males, so the most likely explanation of what they witnessed is that the lack of COUP-TFII protein somehow increased action of testosterone in these genetically modified XX embryos. The researchers ran a number of tests to explore this possibility. Testosterone is mostly produced by the gonads, so they compared the gene expression and enzymes of ovaries of unmodified XX mice with the ovaries of XX mice that lacked the COUP-TFII protein, and they found no differences that pointed to differences in testosterone production. They then considered the possibility that testosterone was produced somewhere else in the body, but the XX mice that lacked the COUP-TFII protein did not have more masculine body features compared to the unmodified XX mice. Finally, the researchers gave extra testosterone to the mother mice that were pregnant with unmodified XX mice and XX mice that lacked the COUP-TFII protein. The extra testosterone did not affect any of the mouse pups; it did not cause the Wolffian ducts of the XX mice that lacked the COUP-TFII protein to regress. Together, the researchers found that no, XX embryos that lack COUP-TFII protein do not have any more testosterone-like activity than their non-genetically modified XX sisters. This means that testosterone alone is not enough to keep Wolffian ducts.

This research has shown us that for the Wolfian ducts to go away during the reproductive development of a mammalian female, they need to be actively dismantled using a biochemical process (similar to how AMH dismantles Müllerian ducts during male reproductive development). COUP-TFII protein appears to be the chemical in charge of triggering this process. Female mammals are not the passive result of simply not becoming male, as has been taught in physiology classes for decades. Becoming a female mammal requires a process all its own, and we are only now starting to learn what that is.


Want to know more? Check these out:

F. Zhao et al. Elimination of the male reproductive tract in the female embryo is promoted by COUP-TFII in mice. Science. Vol. 357, August 18, 2017, p. 717. doi: 10.1126/science.aai9136

A. Swain. Ductal sex determination. Science. Vol. 357, August 18, 2017, p. 648. doi: 10.1126/science.aao2630

The Olympic Athlete of the Animal Kingdom: The Circulatory System of a Horse (A Guest Post)

By Emily Fandrey


How do you judge the abilities of an athlete? Is it all about speed? What about endurance? Strength? How would you judge an animal that can run up to 48 kilometers per hour (30 mph), cover 48 kilometers (30 miles) in a day, or clear a 2.4 meter (8 foot) jump, all while carrying a human on its back? Because of these abilities, the horse (Equus caballus) is often considered to be one of the animal kingdom’s best athletes. The major factor behind horses’ advanced athleticism is their unique circulatory system, specialized for delivering large amounts of oxygen throughout the body.

Image by Paul Kehrer at Wikimedia Commons.

A horse’s circulatory system has three major players: the heart, the spleen, and the frog (and no, this has nothing to do with the animal frog, but rather a specialized unit of a horse’s hoof). Due to these three components, horses have one of the best aerobic capacities in the animal kingdom. Let’s look at a racehorse for example: During a race, a thoroughbred can reach a maximum oxygen capacity (the amount of oxygen the blood can carry) of 200 milliliters per kilogram per minute, meaning 200 milliliters of blood per kilogram of weight (or 3.1 ounces per pound) are transported to the body every minute! This is more than twice the oxygen capacity of the most elite human athlete!

This diagram illustrates the horse’s circulatory system,
including the heart, arteries, veins, and spleen. Diagram by Emily Fandrey.

So let’s break down this superior aerobic system, starting with the horse heart. Typically, a horse’s heart weighs 1% of its total body weight; meaning if a horse weighs 450 kilograms (1000 pounds), its heart will be roughly 4.5 kilograms (10 pounds). If this was true for humans, a 68 kilogram (150 pound) human’s heart would be 0.68 kilograms (1.5 pounds), although the average human heart is only about 0.23 kilograms (half a pound). The horse’s heart functions very similarly to a human heart. It contains four chambers and is responsible for getting oxygen to the body by pumping the oxygen-filled blood. After the body systems have used the oxygen in the blood, this deoxygenated blood enters the heart and is sent to the lungs where the blood is resupplied with oxygen from breathing air. This oxygenated blood reenters the heart and is pumped back out to the body. Because of the size of their hearts, horses are able to supply large amounts of blood with oxygen to the body with each heartbeat, averaging a combined 38 liters (10 gallons) per minute (this is about ten times as much as a human).

Horses also have very different heart rates than humans during rest and exercise. A horse’s resting heart rate is 28-44 beats per minute (bpm), compared to the average human’s, which is 60-80 bpm. During exercise, a human’s heart rate is 90-170 bpm, depending on age. A horse’s heart rate, however, rises to 80 bpm during a walk, 130 bpm during a trot, 180 during a canter, and 240 bpm while galloping. At top speed, the fast beating heart of the horse is what allows the heart to pump much more blood to the body than a human, increasing their athletic abilities.

A diagram of how the horse’s frog sends blood back to heart quickly,
working against gravity. Diagram by Emily Fandrey.

With the long legs of horses, the heart also has to work against gravity to get blood from the limbs back to the heart. To combat this, the horse has its “frog”. For a horse, the frog is a vessel-filled tissue structure on each of its four hooves. When weight is placed on the frog, this structure can help the heart work against gravity. How? When the horse’s hoof meets the ground, the ground will push up on the frog, resulting in the frog being compressed and squeezing blood in the vessels out and rapidly up the leg. The frog helps heart work against gravity by sending the blood up the leg and back to the heart, allowing for faster blood circulation, increasing the athleticism of the horse.

The last key factor to the horse’s circulatory system is the spleen. This organ improves aerobic capabilities and the horse’s athleticism. Now, the primary function of the horse’s spleen is to remove damaged blood cells. However, when a horse is relaxed, their spleen will fill with up to 30 liters (8 gallons) of oxygen-filled blood. And then, once the excitement of activities like running or jumping sparks, the spleen will contract and send up to 25 liters (6.6 gallons) of this stored blood back into circulation in mere seconds! So in seconds, the spleen is capable of almost doubling the maximum amount of oxygen the blood can carry, increasing the athleticism of the horse as well.

So if you ever need an excelling athlete on your team, consider an animal with a superior circulatory system: the horse. With a large and powerful heart capable of pumping large amounts of blood, a spleen to provide an extra burst of blood in seconds, and a “frog” to work against gravity, there is no wonder why horse is considered to be one of the world’s superior athletes.


References

Allen, K.J., Young, L.E., and Franklin, S.H. (2016). Evaluation of heart rate and rhythm during exercise. Equine Veterinary Education 28: 99-112. DOI: 10.1111/eve.12405.

Cardiovascular System (2007). In EQUINAvet.

Circulatory System of the Horse (2010). In Helpful Horse Hints.

Equine Circulatory System Vet, Horse First Aid (2012). In Equestrian and Horse.

Norton, J. (2013). The equine circulatory system. In EquiMed: Horse Health Matters.

Caught in My Web: Animal Reactions to A Solar Eclipse

Animation by Locutus Borg at Wikimedia Commons.

A solar eclipse is a rare event that can have dramatic effects not only on us people, but on animals as well. For this edition of Caught in My Web, we think about how animals may respond to such a rare celestial event.

Image by Luc Viatour at Wikimedia Commons.

1. National Geographic shares many ways animals are known to behave in strange ways in response to a solar eclipse.

2. Much of the effect of an eclipse on animal behavior is utter confusion, but many groups will be watching animals to see how they respond.

3. Researchers at the University of Nebraska will be collecting behavior data on GPS-tagged red-tailed hawks to see how the solar eclipse affects them.

4. Some nature centers are studying animal behavior during the eclipse.

5. If you found any animals that freaked out so badly as to injure themselves, check here for advice.

Drinking Beer Makes You More Attractive… To Mosquitoes

Summer is a time of backyard bar-b-ques, camping, baseball games, beer, and mosquitoes. Ugh, mosquitoes! Have you ever noticed that when a bunch of us are hanging out together outside, some of us get eaten alive by those pesky buggers while others are hardly touched at all? It turns out, differences in how much alcohol we have imbibed may be a factor.

An Anopheles gambiae mosquito ready for a meal. Photo by James D. Gathany
at the Public Health Image Library at Wikimedia Commons.

“No! Say it ain’t so!”

I hate to be the bearer of bad news, so I’ll let the scientific evidence speak for itself.

A research team from the French Research Institute for Development, including Thierry Lefèvre, Louis-Clément Gouagna, Eric Elguero, Didier Fontenille, François Renaud, Carlo Costantini, and Frédéric Thomas, and Kounbobr Roch Dabiré, from the Institute for Research in Health Sciences in Burkina Faso set out to test whether people were more attractive to female mosquitoes after drinking a beer compared to beforehand. They only tested females because only female mosquitoes bite, requiring extra protein for their eggs.

The researchers put groups of 50 hungry female mosquitoes into the end of a special Y-shaped maze that let them fly in the direction of one of two odors. At the end of one arm of the Y-maze was a fan, simply blowing outdoor air through a tent and into the apparatus. At the other end of the Y-maze was a fan blowing air through a tent past a shirtless man and into the apparatus. This shirtless man had either not had anything to drink recently, or had recently drunk either a liter of beer or a liter of water. Between the starting chamber and both ends of the arms of the Y-maze were traps that would capture mosquitoes that had chosen to head that direction (lucky for the shirtless men). The number of mosquitoes caught in both traps combined (compared to the total of 50 that was initially released) was called mosquito activation, and reflected how many mosquitoes were motivated to take off and fly upwind. The proportion of mosquitoes caught in the volunteer-bated trap compared to those caught in both traps combined was called mosquito orientation, and reflected the attractiveness of the volunteer’s odor compared to the control odor.

Image A shows the two tents: one in which the man-bait sat (having consumed beer or water), and the other with no one in it. Air from each tent blew threw a tube (seen in picture B) and then into the building, past the traps and into the downwind box, where the mosquito starting-line was located (seen in picture C). Photos from Lefèvre et al., 2010.

The mosquitoes significantly increased both activation and orientation in response to the beer-drinking volunteers, but not in response to the water-drinking volunteers. That is to say, that the smell of someone that has had a beer motivates more mosquitoes to actively pursue them, and makes them more of a focused target of the mosquitoes. The researchers believe there is an interaction between how our bodies naturally smell and how our bodies break down beer that increases the attractiveness of our odors to mosquitoes. People that were more attractive to mosquitoes before they drank were also more attractive to mosquitoes after they drank. But interestingly, people that were warmer or gave off more CO2 were not more attractive to mosquitoes.

You should know that this research is much more important than just being a drag on your summer bar-b-que. The particular mosquito species that these researchers studied was Anopheles gambiae, the primary vector for malaria in Africa. They did this study in Burkina Faso, a country in West Africa with a high rate of malaria, using a local beer called dolo. Dolo, a fermented sorghum beer with low (3%) alcohol content, is the most common alcoholic beverage in Burkina Faso. So if you are in a place with a high rate of malaria, knowing that you should take extra precautions against mosquitoes when you drink could be a life-saver.

Want to know more? Check this out:

Lefèvre, T., Gouagna, L.C., Dabiré, K.R., Elguero, E., Fontenille, D., Renaud, F., Costantini, C. and Thomas, F. (2010). Beer consumption increases human attractiveness to malaria mosquitoes. PloS one, 5(3), e9546.

How Some Unique Animals Beat the Heat

Man, is it hot out there! While I tuck away in my cool, air-conditioned office, I think about those incredible animals that beat the heat with their own bodies and strategies. It turns out, there are only four major ways to exchange heat with your environment: conduction, convection, radiation and evaporation. The animals that are best equipped for a hot summer are those that take creative advantage of these mechanisms of heat transfer.

"My, what big ears you have!"
"The better to thermoregulate with, my dear!"
Photo by Bernard DUPONT at Wikimedia Commons.

We all exchange heat with our environment wherever our body comes in contact with the environment, namely, across our skin. Animals can increase the rate of this heat-exchange by having a larger surface area relative to the volume of their heat-containing bodies. Big, round animals have a particularly hard time dispersing excess heat, because they have a lot of heat stored in their big bodies, and proportionally not much surface area for the heat to leave from. You may think elephants have such big ears for hearing, but the truth is that they are major temperature-regulation organs. Big, flat body structures provide that added surface area for excess heat to dissipate from. Elephants have large ears with lots of large blood vessels, so they can pump hot blood from the body out to the ears, where they are closer to the cooler environment to dissipate by conduction and convection. The cooler blood then returns to the body core to cool it down.

Built for a life in the cool underground.
Photo by Ted M Townsend at Wikimedia Commons.

Reptiles use a different approach to take advantage of principles of conduction and convection, by burrowing. Burrows create a layer of insulation (usually soil and plant matter) that slows heat exchange and keeps the area below ground a more constant temperature. This means that, compared to the outside, burrows remain cooler in the summer and warmer in the winter. Many reptiles go into their summer burrow in the heat of day and emerge during cooler times of day. Blind lizards, however, take this burrowing idea to an extreme. Blind lizards are a family of legless lizards found in tropical forests. They are small, skinny, and have narrow heads, which make them look more like an earthworm than a lizard, but it also gives them great burrowing efficiency to stay underground and avoid the heat.

A dragonfly exposing as little of his body to the sun
as possible. Photo by Raphael Carter at Wikimedia Commons.

On a hot summer day, we all seek out the shade. This is to reduce the heat we absorb through radiation, and most of this radiation comes from the sun. But what do you do if you can’t find shade? Some dragonflies and damselflies raise their abdomens to aim their rear-ends towards the sun so they can shield themselves from the full-on intensity of the sun’s rays. This body position is called the obelisk posture, because when the sun is directly overhead, the insect’s handstand looks like an obelisk.

Really? Crapping on my own feet?
There has GOT to be a better way.
Photo by Rob Schoenmaker at Wikimedia Commons.

Evaporation is the most efficient way to dissipate heat. Some animals swim, some animals sweat, some animals pant, but vultures and storks win the cooling efficiency award. Vultures and storks. . . poop on their own legs and feet. With this approach, called urohydrosis, the animal releases a mixture of urine and poop through their single excretory hole, called a cloaca, onto their legs and feet. The subsequent evaporation from the high surface area of their long legs helps them to cool off.

So what can we learn from these heat-beating experts (without pooping on ourselves)? If you are hot, spread your body out, stay in the shade, and wet an area of your body with high surface area and exposed blood vessels (namely, your inner wrists and forearms). I’d use water though.