The idea of sex differences in the brain both fascinates and inflames the public. As a result, the communication and public discussion of new findings is particularly vulnerable to logical leaps and ...pseudoscience. A new US National Institutes of Health policy to consider both sexes in almost all preclinical research will increase the number of reported sex differences and thus the risk that research in this important area will be misinterpreted and misrepresented. In this article, I consider ways in which we might reduce that risk, for example, by (i) employing statistical tests that reveal the extent to which sex explains variation, rather than whether or not the sexes ‘differ’, (ii) properly characterizing the frequency distributions of scores or dependent measures, which nearly always overlap, and (iii) avoiding speculative functional or evolutionary explanations for sex-based variation, which usually invoke logical fallacies and perpetuate sex stereotypes. Ultimately, the factor of sex should be viewed as an imperfect, temporary proxy for yet-unknown factors, such as hormones or sex-linked genes, that explain variation better than sex. As scientists, we should be interested in discovering and understanding the true sources of variation, which will be more informative in the development of clinical treatments.
As part of an initiative to improve rigor and reproducibility in biomedical research, the U.S. National Institutes of Health now requires the consideration of sex as a biological variable in ...preclinical studies. This new policy has been interpreted by some as a call to compare males and females with each other. Researchers testing for sex differences may not be trained to do so, however, increasing risk for misinterpretation of results. Using a list of recently published articles curated by Woitowich et al. (eLife, 2020; 9:e56344), we examined reports of sex differences and non-differences across nine biological disciplines. Sex differences were claimed in the majority of the 147 articles we analyzed; however, statistical evidence supporting those differences was often missing. For example, when a sex-specific effect of a manipulation was claimed, authors usually had not tested statistically whether females and males responded differently. Thus, sex-specific effects may be over-reported. In contrast, we also encountered practices that could mask sex differences, such as pooling the sexes without first testing for a difference. Our findings support the need for continuing efforts to train researchers how to test for and report sex differences in order to promote rigor and reproducibility in biomedical research.
Biomedical research has a long history of including only men or male laboratory animals in studies. To address this disparity, the United States National Institutes of Health (NIH) rolled out a policy in 2016 called Sex as a Biological Variable (or SABV). The policy requires researchers funded by the NIH to include males and females in every experiment unless there is a strong justification not to, such as studies of ovarian cancer. Since then, the number of research papers including both sexes has continued to grow.
Although the NIH does not require investigators to compare males and females, many researchers have interpreted the SABV policy as a call to do so. This has led to reports of sex differences that would otherwise have been unrecognized or ignored. However, researchers may not be trained on how best to test for sex differences in their data, and if the data are not analyzed appropriately this may lead to misleading interpretations.
Here, Garcia-Sifuentes and Maney have examined the methods of 147 papers published in 2019 that included both males and females. They discovered that more than half of these studies had reported sex differences, but these claims were not always backed by statistical evidence. Indeed, in a large majority (more than 70%) of the papers describing differences in how males and females responded to a treatment, the impact of the treatment was not actually statistically compared between the sexes. This suggests that sex-specific effects may be over-reported. In contrast, Garcia-Sifuentes and Maney also encountered instances where an effect may have been masked due to data from males and females being pooled together without testing for a difference first.
These findings reveal how easy it is to draw misleading conclusions from sex-based data. Garcia-Sifuentes and Maney hope their work raises awareness of this issue and encourages the development of more training materials for researchers.
•More than 50 variants of sex steroid receptor genes are associated with human behavioral phenotypes.•Most of the polymorphisms do not occur in coding regions, making interpretation ...challenging.•Variation in non-coding regions can affect gene transcription via a variety of mechanisms.•Making use of in vitro assays and non-human models will advance understanding of these processes.
Sex steroid receptors have received much interest as potential mediators of human behaviors and mental disorders. Candidate gene association studies have identified about 50 genetic variants of androgen and estrogen receptors that correlate with human behavioral phenotypes. Because most of these polymorphisms lie outside coding regions, discerning their effect on receptor function is not straightforward. Thus, although discoveries of associations improve our ability to predict risk, they have not greatly advanced our understanding of underlying mechanisms. This article is intended to serve as a starting point for psychologists and other behavioral biologists to consider potential mechanisms. Here, I review associations between polymorphisms in sex steroid receptors and human behavioral phenotypes. I then consider ways in which genetic variation can affect processes such as mRNA transcription, splicing, and stability. Finally, I suggest ways that hypotheses about mechanism can be tested, for example using in vitro assays and/or animal models.
Measuring stress in animals is important in many ecological, zoological and veterinary research settings. A common method is to measure plasma levels of glucocorticoid hormones (cortisol, ...corticosterone, hereafter CORT). Over the past decade, an alternative method has become widely popular: assessing leucocyte profiles; in other words, the heterophil‐to‐lymphocyte ratio (H/L) in birds and reptiles or the neutrophil‐to‐lymphocyte ratio (N/L) in mammals, amphibians and fish. Recent studies have shown that although both indices reliably change after acute stress, they are not always correlated at baseline, and thus are not interchangeable. This lack of correlation has led to some confusion about which of these two measures is “better”; in other words, more biologically relevant and a truer readout of stress.
In this review, we first document the dramatic expansion of the use of leucocyte profiles to assess stress over the past decade. We then review published evidence that may explain the discrepancies between leucocyte profiles and plasma CORT. We argue that these discrepancies stem from the nature of the stress reaction and the role of corticosterone (or cortisol) versus leucocytes during the stress response.
The differing roles of the two responses translate to differences in the timing of each, which is important for researchers studying either acute or chronic stress. Although plasma glucocorticoids rise within minutes of the onset of acute stress and return to baseline within 1 or 2 hr after the stressor passes, H/L or N/L ratios remain low for at least 30–60 min or sometimes more.
The temporal nature of these metrics differs also in the context of long‐term chronic stress. The glucocorticoid response wanes in repeatedly stressed animals, or those exposed to chronic environmental stress, which can often (but not always) lead to low baseline levels and more typically, an attenuated acute stress response. Meanwhile, H/L or N/L ratios appear to remain high under the same conditions.
Understanding the temporal differences in the responses should help researchers decide which metric is the better method in their particular study, or whether both would be useful for elucidating different elements of the stress response.
Research highlights ► Estradiol levels are regulated by auditory experience and modulate auditory-based behaviors in songbirds. ► Brain-generated estradiol enhances the gain and auditory coding ...efficiency of central auditory neurons in real-time. ► Estradiol suppresses local GABAergic transmission in central auditory neurons via a pre-synaptic, non-genomic mechanism. ► Estradiol is necessary and sufficient for the induction of plasticity-associated genes in central auditory neurons. ► Catecholaminergic transmission may regulate estradiol’s modulation of auditory processing and behavior.
Biomedical research has a history of excluding females as research subjects, which threatens rigor, reproducibility, and inclusivity. In 2016, to redress this bias, the U.S. National Institutes of ...Health (NIH) implemented a policy requiring the consideration of sex as a biological variable (SABV) in all studies involving vertebrate animals, including humans. Unless strongly justified, females and males must be included in all studies and results reported disaggregated by sex. Recent evidence indicates, however, that misunderstandings of the policy and other significant barriers impede its implementation. To shed light on those barriers at our home institution, we conducted a study funded by the Emory University Specialized Center of Research Excellence on Sex Differences (SCORE). In semistructured interviews of Emory principal investigators in the biological sciences, we noted their knowledge of what the policy entails and why it was implemented, their attitudes toward it, and the extent to which it has or has not changed their research practices. Although attitudes toward SABV were generally positive, most researchers face challenges with respect to its implementation. We suggest interventions that can be mounted at the level of home institutions, such as raising awareness of locally available core facilities, to help address these challenges. More training is needed on what the policy asks of researchers, how sex is defined, the nonhormonal ways that sex differences can manifest, and best practices for statistical analysis of sex-based data. Home institutions may also want to explore ways to lessen the stress associated with rollout of SABV policy.
To enhance inclusivity and rigor, many funding agencies and journals now mandate the inclusion of females as well as males in biomedical studies. These mandates have enhanced generalizability and ...created unprecedented opportunities to discover sex differences. However, education in sound methods to consider sex as a subgroup category has lagged behind, resulting in a problematic literature in which study designs, analyses, and interpretations of results are often flawed. Here, we outline best practices for complying with sex-inclusive mandates, both for studies in which sex differences are a primary focus and for those in which they are not. Our recommendations are organized within the "4 Cs of Studying Sex to Strengthen Science: Consideration, Collection, Characterization and Communication," a framework developed by the Office of Research on Women's Health at the National Institutes of Health in the United States. Following these guidelines should help researchers include females and males in their studies while at the same time upholding high standards of rigor.
Conspecific vocalizations differ from many other sounds in that they have natural incentive salience. Our thinking about auditory responses to vocalizations may therefore benefit from models ...originally developed to understand reward. According to those models, the brain attributes incentive salience to rewarding stimuli via the activity of monoaminergic neuromodulators. These neuromodulators, in turn, mediate the effects of experience and internal state. Songbirds lend themselves well to this discussion because the natural incentive salience of song is clearly modulated by both factors. Their auditory responses have been well-studied, particularly the song-induced expression of plasticity-associated genes such as ZENK. Here I review evidence that ZENK responses to song are regulated by monoamine neuromodulators, and I interpret this evidence in the context of incentive salience. First, hearing conspecific song engages monoaminergic activity in the auditory system and elsewhere. Second, in females this activity may be regulated by the same hormones that regulate behavioral preferences for song. Finally, much of the evidence thought to implicate neuromodulators in song discrimination and memory suggests that they may affect incentive salience. Expanding the study of incentive salience beyond the mesolimbic reward system may reveal some new ways of thinking about its underlying neural basis.
This article is part of a Special Issue entitled “Communication Sounds and the Brain: New Directions and Perspectives”.
•The processing of vocalizations is considered in the context of incentive salience.•The concept of incentive salience extends beyond the mesolimbic reward system.•In songbirds, hearing song engages monoaminergic activity.•This activity is modulated by associative learning and internal state.•Associative learning and internal state can confound study designs.
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