Can prenatal stress be reversed?

I was scanning the titles of new journal articles a while back, and came across one that made me think, hey, that may be about rats, but it is totally relevant to dogs. And then I thought, why don’t I teach a class on it? Read and interpret this really interesting journal article with a group of dog trainers and dog lovers?

I will be teaching the class Prenatal Stress and Anti-Depressants for APDT the week of November 18 (and you are invited to take it). This post will be used as reading material for it. In the class, we will talk about this article and what conclusions we can draw from it and apply to dogs. So I may not draw as many the conclusions for you in this post as I usually do; the plan is for the students to do that together in class. But it was a fascinating paper and there’s lots of good material in it, so read on if you want a conclusion-free summary of it!

Pereira-Figueiredo I., Juan Carro, Orlando Castellano & Dolores E. Lopez (2014). The effects of sertraline administration from adolescence to adulthood on physiological and emotional development in prenatally stressed rats of both sexes, Frontiers in Behavioral Neuroscience, 8 DOI: http://dx.doi.org/10.3389/fnbeh.2014.00260


Why prenatal stress?
So what’s prenatal stress and why is it important to dogs? The authors of the article don’t provide much background on this phenomenon, but it’s an interesting one: when a mammal undergoes unusually high stress during her pregnancy, the personality of her offspring can be affected. We believe that the stress hormones rising in her bloodstream can pass through her placenta to the fetus or fetuses, and can change how their brains develop at this very early stage of life. That’s prenatal stress: stress experienced before birth.

Part of what this paper investigates is exactly how prenatal stress affects the developing personality, because we don’t yet fully understand how this stuff works. In general, though, we expect prenatally stressed animals to be more anxious and less confident than animals who were not prenatally stressed.

Does prenatal stress affect dogs? We don’t know for sure, but we think it is something that can affect most or all mammals. Would it happen commonly? Hard to say, but I imagine a pregnant dog who is stray or in a shelter and highly stressed, and I wonder what effects this might have on her puppies.

What can you do about it?
If you have a puppy that you believe was prenatally stressed and whose personality you thought was adversely affected, what are your options? Careful socialization and good enrichment are always an excellent choice, but in some cases you might consider medication. This study looks at whether a particular anti-depressant, sertraline, might help change the individual’s personality long-term if given in adolescence. Sertraline is an SSRI, in the same class of medications as Prozac. These are widely used medications believed to be fairly safe, but one of the questions these researchers ask is whether it is safe when given throughout an animal’s entire adolescence.

SSRIs such as sertraline affect the levels of serotonin in your brain. Serotonin is a chemical which affects mood; depressed people tend to have less of it, as do aggressive people. As a result, it is the target of a fair number of anti-depressants, which work to increase its levels. Prenatal stress is known to disrupt the serotonin system in the brain, so a medication which affects serotonin is a reasonable choice for prenatally stressed individuals.

So the idea is: give these prenatally stressed animals a medication which increases their serotonin levels while they are adolescents and their brains are still developing. The hope is that they will develop into more normal adults than they would have without the medication. So, exactly how do you investigate such a question?

Methods: how the study worked
First, the researchers stressed some pregnant rats by putting them in clear tubes to restrain them, and shining bright light on them. This was repeated for forty-five minutes at a time, three times a day. They also kept control rats, who were not stressed during their pregnancy. The pups born to these two sets of rats were then in two categories: prenatally stressed pups and non-stressed pups.

The rat pups began their anti-depressant treatment with sertraline when they were one month old. Now there were four groups of rat pups:

prenatal stress anti-depressants	prenatal stress no anti-depressants
no prenatal stress anti-depressants	no prenatal stress no anti-depressants

Having these four groups allowed the researchers to pick apart the two different effects, the effect of prenatal stress and the effect of anti-depressants during adolescence.

The pups were tested at two months of age, the beginning of rat adolescence, to see if the prenatal stress had affected their personalities. They were assessed for how they dealt with startling noises and being exposed to open space (scary for a rat!). Their blood was also tested to see how their immune systems were developing, because immune systems develop differently in animals who have been subjected to high stress. All these tests were run again one month later, at the end of rat adolescence, to see how the anti-depressant given throughout adolescence had affected treated rats compared to the control groups.

Results: what they found

• Although we may think of prenatal stress as mostly affecting an animal’s behavior, it’s been shown to also affect metabolism, so this study looked at birth weight. Interestingly, prenatal stress only reduced the birth weight of the female rat pups, not the males. The weights of these females had equalized compared to non-stressed pups by weaning age. After weaning, though, the prenatally stressed females continued to gain weight and ended up heavier as adults than the non-stressed females. When prenatally stressed females were given the anti-depressant sertraline, however, this weight difference was reduced.
• The pups were also tested for their startle response when they heard a sudden sound. Prenatally stressed rat pups did seem to have a larger startle amplitude (size) compared to controls, but this wasn’t statistically significant, and was not reversed by sertraline treatment.
• Prenatally stressed females did not habituate to the startling sound after several exposures as well as rats from other groups did; treatment with sertraline reversed this effect.
• The pups’ behavior in an open space was tested. No difference was seen between prenatally stressed and non-stressed pups, except in males on their first time being tested (not on later tests).
• In the open space test, only non-stressed females explored more (became more confident) on repeated testing; males and prenatally stressed females did not become more confident with repeated exposure to the open space.
• The pre-natally stressed rats showed a significant decrease in their number of white blood cells. This change was reversed when they were treated with sertraline.

Discussion: what does it all mean?
The study’s main conclusions are that effects of prenatal stress can be seen in rats, and that giving sertraline during adolescence did not harm them.

The rapid weight gain in the pre-natally stressed females is an effect that’s been seen before, and seen in humans. Children born with low birth weights often grow to have issues with their weight and can suffer from diseases related to a poorly regulated metabolism. This loss of control of energy balance has been associated with dysregulation of serotonin in humans, adding additional support to the choice of sertraline, an anti-depressant which interacts with the serotonin system.

It is interesting that no anxiety-like behavioral changes were seen in the prenatally stressed rats. Prenatal stress is known to cause anxious personalities in many cases. However, these rats were as confident (or as anxious!) in the open space test as rats who had not been prenatally stressed. The researchers comment that this particular test has been done on prenatally stressed rats in other studies, and that those rats didn’t show anxiety in the open space test either, so this does seem to be a real result, rather than a statistical error.

They did see some differences, though. Male rats who had been prenatally stressed did show some additional reluctance to explore (i.e. anxiety) on their first day only in the open space test. After that they explored equal amounts compared to other groups.

The researchers also note that while most of the rats that they tested were equally anxious on all days that they were tested in the open space, females who had not been prenatally stressed appeared to begin to explore more on repeated tests, as if they were learning to be less anxious as their surroundings became more familiar. This was not the case in male rats or in rats who had been pre-natally stressed.

Remember also that female rats who were prenatally stressed did not habituate to startling sounds as well as rats from other groups. Is it possible that with this particular model of prenatal stress, the personality effects of prenatal stress appear not as classical anxiety, but as difficulty habituating to new situations or stressors such as loud noises?

Finally, the researchers looked at effects of pre-natal stress on the immune system, and found significant effects (decreased numbers of white blood cells) which were reversed by treatment with the anti-depressant sertraline. Why did they care about the immune system? Because the immune system and the stress system are closely intertwined. Stressed animals show changes in the numbers of their white blood cells just as the prenatally stressed rats did. The researchers were using the changes in the immune system as markers for changes in the stress system.

There are two possibilities for why these prenatally rats showed stress-associated changes in their immune systems: either because they themselves had high stress levels, or because their immune systems were developing prenatally (in utero) in a high stress environment due to their mother’s stress levels. Either way, it is interesting that treatment with sertraline reversed these effects, suggesting that it may have either changed current stress levels in these adolescent rats (even though the only serious stressors they had undergone were before their birth!), or had counteracted other effects from that prenatal stressor.


Conclusions
It can be hard to know exactly what conclusions to draw from a scientific paper. What do you think? What are the most important findings in this paper (maybe just two or three of them)? Do you think those findings are real phenomena, or maybe just statistical mistakes? If they’re real, do you think they can be extrapolated from rats to humans or dogs?

Domestication and human evolution, streaming!

Domesticated humans, domesticated canids, domesticated finches! The Center for Academic Training & Research in Anthropogeny held a conference on domestication and human evolution, and live streamed it. I watched live, refusing to speak to my husband or dogs during it, which didn’t go over well. You, however, can watch the archive on YouTube.

The conference was a series of short talks from researchers. These included:

Robert Wayne, “The transformation of wolf to dog: history, traits, and genetics”

Wayne’s lab published a recent genetics paper on exactly where the dog was domesticated, and in this talk he stepped us through their findings. Previous work (and there has been plenty of it) on the location of dog domestication has begun with the assumption that dogs evolved from a population of wolves which still exists basically unchanged, inhabiting the same range now as it did then. Previous work has suggested that this occurred in Asia or the Middle East. Wayne’s group argues that the population of wolves which gave rise to dogs no longer exists, but lived in Europe about 20,000 years ago. It’s a different perspective on a question which is very difficult to answer, because dogs continue to interbreed with wolves so freely that these genetic studies are awfully hard to interpret.

Anna Kukekova, “Fox domestication and the genetics of complex behaviors”

This talk came out of the lab where I work and I got to see my own name on the list of contributors at the end of the talk. Kukekova gave an overview of the history of the fox domestication project, in which lines of foxes were selectively bred for tame temperament or aggressive temperament, and recent research. Our lab digs in to the question of what it is in the genetics of the tame foxes that makes their personalities so different from those of conventional farm foxes. Since this conference was about domestication, and the tame foxes are the best known and longest running domestication study, speakers returned to the foxes throughout their talks. They are a tough nut to crack. Behavior is exceedingly complex mechanistically and we (by which I mean all behavioral geneticists, not just our lab) are still trying to figure out how to get a handle on the genetics that affect it.

Robert Franciscus, “Craniofacial feminization in canine and human evolution”

Craniofacial feminization means that your face is flat, basically. Look at the reconstruction of a Neanderthal face: the chin juts out. Look at a modern human face: flat. Look at a chimpanzee face: jutting chin. Look at a baby chimpanzee: flat. Do humans look like baby chimps? We kind of do. Is there a significance to this? Franciscus argued convincingly that there is, and discussed differences in dog versus wolf muzzle length (and got quite technical about how his group investigated them). We don’t know why this feminization or neotenization (childlike changes) happens in domestication, but it seems to be a recurrent theme. This was the first talk that really grappled with the idea that humans are domesticated, with changes compared to our recent ancestors that parallel changes between dogs and wolves, or between tame and conventional foxes.

Terrence W. Deacon, “The domesticated brain”

Do domesticated animals have smaller brains than their wild counterparts? This is certainly the case in dogs and wolves. Is it the case in humans and our ancestors? Deacon’s group has studied Neanderthal brain size based on their skulls, and they conclude that modern humans do not have clearly smaller brains. He noted that the tame foxes also do not appear to have smaller brains than their conventional counterparts. Why does the difference in brain size show up in only some, but not all, examples of domestication? Is it perhaps not a necessary part of the domestication process?

Phillipp Kaltovich, “Neotenous gene expression in the developing human brain”

It is pretty difficult to study gene expression in human brains, because you have to cut up brains to get your data. Kaltovich did get his data from somewhere, though, and it was really interesting to see his comparisons of gene expression in young versus older brains. The question was whether gene expression changes with age, which helps get at the bigger question, are there gene expression differences in domesticated species compared to their wild counterparts, and are these expression differences similar to the differences in young versus mature animals? In other words, are domesticated species basically enternally young (neotenized)? He did find differences, but his group will have a long way to go to put them together into findings that really illuminate the domestication question. I have a lot of sympathy for how hard this particular approach is, as my research is currently also focused on brain gene expression.

Tecumseh Fitch, “The domestication syndrome and neural crest cells: a unifying hypothesis”

In a recent paper, Fitch’s group put forth the concept of a domestication syndrome, a set of changes associated with domestication: flatter face, behavioral changes, white markings, etc. Subsets of these changes are seen in all domesticated species. Fitch’s group hypothesizes that a particular kind of cell involved in early development is involved in all of these changes. This cell, the neural crest cell, migrates through the growing embryo and develops into many different structures and cell types, including coloration cells (explaining white markings), teeth (explaining dentition changes), and the adrenal medulla (source of adrenaline, explaining behavioral changes). It’s an interesting hypothesis and I’ll be curious to see where this group goes with validating it.

Kazuo Okanoya, “Domestication and vocal behavior in finches”

Okanoya’s group studies a species of domesticated finch and its very closely related wild ancestor. The wild finch has a simple song, while the domesticated species has a quite complicated one. Okanoya’s group investigates the difference in these songs, as a model for the development of complex language in domesticated humans. He played both songs, wild and domesticated, and the difference was dramatic. He linked the changes in the song between species to sexual selection.

Richard Wrangham, “Did Homo sapiens self domesticate?”

The question of self domestication was one of the recurring themes of the conference, and for me this was the most transfixing session. Wrangham studies chimpanzees and bonobos, two closely related species with very different aggression levels, as models of the difference between humans and our more aggressive, non-domesticated ancestors. He defines domestication as the reduction of reactive aggression. Reactive aggression is different from proactive aggression: humans are quite good at controlling our reactive aggression, as we are able to tolerate strangers and live in large groups very well. But we do still show significant proactive aggression, which he described as aggression performed in cold blood, such as armed robbery. Wrangham suggests that a reduction in aggression is the trait evolutionarily selected for in self-domestication, and the other parts of the package (flatter faces, white markings) come along for the ride as associated traits. Self-domestication is often seen in island species, and he gave the example of the red colobus with a neotonous (childlike) island version compared to the mainland population.

The whole conference was really fascinating. It’s available on YouTube now, so go, check it out!

State of the Zombieverse

I haven’t been blogging much lately, and it’s mostly because I’ve been writing so much for more mainstream media outlets:

• “Neutering without a Scalpel” in the summer issue of the Whole Dog Journal. This story was about Zeuterin, a new product for performing chemical castration on a dog — in other words, non-surgical neutering. I tried to cover all the possible pros and cons of using Zeuterin versus the traditional surgical approach. I don’t think either solution is going to be right for every dog or every situation, and it’s nice to see new options coming out.
• “Testing the Tests“ in the fall issue of The Bark. I read so many journal articles about canine behavioral assessments (also known as temperament tests) for this piece. You know, the tests that are often used in shelters to try to identify behavior problems in dogs before they’re put up for adoption. It was fascinating reading and ended up telling an interesting story about researchers’ attempts to figure out whether behavioral assessments are good at predicting canine behavior. The answer: not really, but some new research approaches have appeared recently which have shed a lot of light on how to interpret these tests and how to approach building better tests.
• I’ve also been writing for DVM360 (I have two pieces going through the pipeline there right now, one about shelters and one about stress), I have another piece pending for the Whole Dog Journal, a guest post for a new dog site, and you already know about my upcoming ultra short, ultra fun online class for APDT, right?

So with all of that, I haven’t been blogging much, but I keep thinking about it and missing it. I figured I should at least report here on where to find me.

Journal articles, stress, and anti-depressants with me and APDT

I’m teaching another online course for APDT: Analyzing Journal Articles: Pre-Natal Stress and Anti-Depressants. I’m trying something new with this one. It's just one week long. Basically, I’m going to be walking through a recent journal article that I thought had interesting implications for dogs. There will be several short lectures at different experience levels. Some will provide background in the area to students who don’t have an extensive science background. Some will be aimed at students who do know a lot of science and want to dig deeper into stuff that isn’t often covered in online courses. And some will be straightforward “what is this article about?”

It's an interesting article, about whether stress before birth can affect an animal’s personality and whether anti-depressant medication change reverse those effects. Hopefully it isn’t hard to see how this could be relevant to dogs with behavior issues.

If we don't get at least ten students signed up, we won’t do it, so go sign up now! You will get CE! If people do sign up and do enjoy it, I’m hoping to do more like this -- providing a way for dog trainers to keep up on recent research and get a better feel for what it’s like to read a scientific paper. So this is my test case. Will people be interested? I really hope so!

[ETA: Although the last time I checked, the APDT web page for this class lists it as "no CEUs," I checked with their education coordinator, and students completing this class will earn 4 CEUs. So yes, you will get CEU credit for it!]