New research finds caffeine consumed during pregnancy can change important brain pathways that could lead to behavioral problems later in life.
Researchers in the Del Monte Institute for Neuroscience at the University of Rochester Medical Center (URMC) analyzed thousands of brain scans of nine and ten-year-olds, and revealed changes in the brain structure in children who were exposed to caffeine in utero.
“These are sort of small effects and it’s not causing horrendous psychiatric conditions, but it is causing minimal but noticeable behavioral issues that should make us consider long term effects of caffeine intake during pregnancy,” said John Foxe, Ph.D., director of the Del Monte Institute for Neuroscience, and principal investigator of the Adolescent Brain Cognitive Development or ABCD Study at the University of Rochester.
“I suppose the outcome of this study will be a recommendation that any caffeine during pregnancy is probably not such a good idea.”
“What makes this unique is that we have a biological pathway that looks different when you consume caffeine through pregnancy,” said Zachary Christensen, a M.D/Ph.D. candidate in the Medical Science Training Program and first author on the paper published in the journal Neuropharmacology.
“Previous studies have shown that children perform differently on IQ tests, or they have different psychopathology, but that could also be related to demographics, so it’s hard to parse that out until you have something like a biomarker.
This gives us a place to start future research to try to learn exactly when the change is occurring in the brain.”
Investigators analyzed brain scans of more than 9,000 nine and ten-year-old participants in the ABCD study. They found clear changes in how the white matter tracks – which form connections between brain regions – were organized in children whose mothers reported they consumed caffeine during pregnancy.
URMC is one of 21-sites across the country collecting data for the ABCD study, the largest long-term study of brain development and child health. The study is funded by the National Institutes of Health. Ed Freedman, Ph.D., is the principal investigator of the ABCD study in Rochester and a co-author of the study.
“It is important to point out this is a retrospective study,” said Foxe. “We are relying on mothers to remember how much caffeine they took in while they were pregnant.”
Previous studies have found caffeine can have a negative effect on pregnancy. It is also known that a fetus does not have the enzyme necessary to breakdown caffeine when it crosses the placenta.
The researchers point out that it is unclear if the impact of the caffeine on the fetal brain varies from one trimester to the next, or when during gestation these structural changes occur.
“Current clinical guidelines already suggest limiting caffeine intake during pregnancy – no more than two normal cups of coffee a day,” Christensen said.
“In the long term, we hope to develop better guidance for mothers, but in the meantime, they should ask their doctor as concerns arise.”
Adult brain structure is primarily established in early life (Lenroot and Giedd, 2006). Disturbances in anatomical (number and connectivity of neurons) and functional (ability to engage alternative brain networks) development can interfere with crucial processes, including cell proliferation, neuronal migration (Crandall et al., 2004) and post-migration cortical connectivity (Guerrini and Dobyns, 2014).
Disturbances can be genetic (e.g., a mutation) and environmental (e.g., maternal separation), resulting in increased risk to develop pathologies during early life such as cortical malformations, developmental delay, epilepsy, and/or autism (Barkovich et al., 2012; Guerrini and Dobyns, 2014), as well as during adulthood/aging, leading to dementia such as Alzheimer’s Disease (AD)-related pathology (Borenstein et al., 2006; Whalley et al., 2006; Stern, 2012; Seifan et al., 2015).
Neuropathologically, AD is defined by the extracellular accumulation of amyloid beta (Aß) peptides into amyloid plaques, and the presence of intraneuronal fibrillar aggregates of hyper- and abnormally-phosphorylated tau proteins (Masters et al., 1985; Sergeant et al., 2008).
Tau pathology is observed early in the brain stem and entorhinal cortex (Braak et al., 2011) and its progression from entorhinal cortex, to the hippocampus, and finally neocortex corresponds to the progression of the symptoms in AD (Duyckaerts et al., 1997; Grober et al., 1999) supporting a pivotal role of Tau pathology in AD-related memory impairments.
Various genetic and environmental risk factors are associated with dementia and/or AD (Reitz et al., 2011). Most of these environmental and lifestyle-related factors also impact AD lesions, and in particular Tau pathology. For instance, physical exercise (Belarbi et al., 2011), anesthetics (Le Freche et al., 2012; Whittington et al., 2013), or obesity/diabetes (Leboucher et al., 2013; Papon et al., 2013) modulate Tau pathology and associated memory disturbances.
Among the numerous existing environmental risk factors, those occurring during pregnancy and lactation can have important functional consequences. Exposure to psychoactive substances in utero can alter fetal brain development, leading to pathological states later in life for the offspring, including psychiatric disorders (Marroun et al., 2015; Skorput et al., 2015).
Caffeine is the most frequently consumed psychoactive substance, including during pregnancy (Mandel, 2002; Greenwood et al., 2014). In mice, caffeine exposure during pregnancy and until weaning delays the migration and integration of GABA neurons, enhances seizure susceptibility, as well as alters brain rhythms and hippocampus-dependent memory function in the offspring (Silva et al., 2013; Fazeli et al., 2017).
Although it is difficult to generalize rodent studies to humans, a study in mother–child pairs showed an association between caffeine exposure during pregnancy and impaired cognitive development (Galéra et al., 2015). Guidelines for pregnant women recommend to limit the amount of caffeine consumption to 200–300 mg/kg (American College of Obstetricians and Gynecologists, 2010). Whether early life exposure to caffeine may prime exposed offsprings to the development of neurodegenerative disorders later in life remains unknown.
In the present study, we specifically aimed at determining whether Tau pathology related pathological traits would appear sooner in animals exposed to caffeine during brain development. To address this question, we evaluated the effects of early life caffeine exposure in offspring of the THY-Tau22 transgenic mouse model that progressively develops AD-like hippocampal Tau pathology, with ongoing deficits at 6–8 months of age and a full pathology and memory impairments occurring at 12 months of age (Van der Jeugd et al., 2013).
Psychoactive drugs ingested during pregnancy can have widespread deleterious effects in the fetus, in particular in the brain (Salisbury et al., 2009). Substances of abuse, such as alcohol, cannabis and cocaine, can directly alter the construction of the brain, in particular GABAergic circuits (Miller, 1986; Berghuis et al., 2007; Navi-Goffer and Mulder, 2009; Thompson et al., 2009).
As the most widely consumed psychoactive drug, including during pregnancy, caffeine can also have body-wide effects (Temple et al., 2017). Previous works in control GIN (GFP-expressing Inhibitory Neurons) mice have shown that caffeine slows down the migration of GABA neurons resulting in a delayed insertion in the circuitry, both in the hippocampus and the cortex during the first postnatal week (Silva et al., 2013; Fazeli et al., 2017).
Such alterations may have a direct impact on hippocampal network activity and performance during development (Salesse et al., 2011). Indeed, they are associated with enhanced sensitivity to epilepsy, hyperactivity in vivo and cognitive deficits later in life in offspring (Silva et al., 2013; Fazeli et al., 2017).
Based on these studies, we hypothesized that the modifications produced by early life exposure to caffeine (acting as a first hit) would leave a long-term trace in the circuits, rendering them more vulnerable to a second hit, in line with the diathesis-disease theory (Bernard, 2016).
This theory has been validated in the context of stress-induced vulnerability to epilepsy, depression and cognitive deficits, with the demonstration that social defeat (first hit) induces a state of vulnerability in some rats; a second hit being necessary to trigger a phenotype in the vulnerable population (Blugeot et al., 2011; Becker et al., 2015, 2019; Bouvier et al., 2016).
If early-life exposure to caffeine constitutes a first hit and induces a state of vulnerability to a tauopathy, we reasoned that Tau mutant offspring exposed to caffeine would express phenotypic traits earlier than Tau mutants on water, in particular the cognitive deficits and biochemical alterations known to occur in this mouse model.
Since we wanted to bridge behavioral and molecular levels, we looked at the circuit level, focusing on the CA1 region, which is characterized by strong alterations in the GABAergic and glutamatergic circuits in caffeine exposed WT mice (Silva et al., 2013; Fazeli et al., 2017). Our ultimate goal was to be able to provide a coherent picture, linking all levels of analysis. However, the three levels of analysis do not provide a coherent picture.
The most salient feature of the results is the occurrence of deficits in spatial memory and learning already at 8 months in caffeine exposed Tau mice. These deficits occurred at 12 months in water exposed Tau mice and were still present at that age in caffeine Tau mice.
In the present work, we used the Barnes maze, which is a terrestrial version of the Morris water maze and which is less stressful to mice than the Morris water maze (Harrison et al., 2009). We did not use other tests (such as anxiety and novel object recognition) to prevent interference when using different behavioral tests in succession. Interestingly, caffeine treatment did not alter spatial memory as assessed with the Barnes maze in Wild type mice, whilst caffeine-treated WT mice showed deficits in the object-location memory task (Silva et al., 2013).
This suggests that specific types of memories are affected. The Barnes maze test is particularly useful in our case as WT mice exposed to caffeine do not display deficits, which would have added an independent variable, rendering the interpretation of the results more difficult. We can thus propose that early life exposure to caffeine accelerates the occurrence of the behavioral phenotype (with the caveat that we only looked at two time points).
Humans have different genetic backgrounds and go through different life experiences, which will determine their sensitivity to the development of diseases, in particular neurological disorders (Bernard, 2016). The fact that caffeine exposure during early life produces an earlier occurrence of cognitive deficits in the Tau model used here may be relevant to a subset of human individuals.
Said differently, exposure to caffeine during pregnancy may sensitize some but not all offsprings. Future studies will need to investigate other time points during aging, other cognitive tests and other models of AD. The experimental protocol is however highly time-consuming. We cannot rely on established aging colonies, since females need to be exposed to caffeine in the drinking water before mating until weaning. Then, it is necessary to wait until offspring reach the appropriate age.
Multiple mechanisms can be proposed to explain cognitive deficits in the Barnes maze (activation of the HPA axis, inflammation, metabolic defect, cell death, synaptopathy, channelopathy, epigenetic modifications, to name but a few). In THY-Tau22 mice, cognitive deficits have been associated with Tau pathological load and neuroinflammation (Van der Jeugd et al., 2013; Laurent et al., 2016, 2017, but see Burlot et al., 2015; Chatterjee et al., 2018).
However, analysis of cardinal pTau and neuroinflammatory markers did not correlate with memory deficits in offspring exposed to caffeine. This result is in keeping with the proposal that cognitive decline may occur before the expression of molecular phenotypic traits in patients with AD (Jessen et al., 2014).
Since the hippocampus plays a key role is spatial memory, we were expecting circuit alterations in this region. To assess them, we measured glutamatergic and GABAergic synaptic currents received by hippocampal CA1 pyramidal cells. The results are very difficult to interpret because of the presence of three independent variables imposed by the experimental protocol.
The first two independent variables are caffeine and Tau, both of which result in morpho-physiological alterations. The third independent variable is age, since we are considering two time points, 8 and 12 months. Yet, the in vitro approach provides interesting results in their complexity. Since our hypothesis was that caffeine exposure accelerates the occurrence of the phenotype, we start to discuss the age factor.
In a given condition (WT mice or Tau mice), we were expecting to find whichever properties identified at 12 months in water treated animals at 8 months in caffeine exposed animals. In WT mice on water, we found an increase in both glutamatergic and GABAergic drives between 8 and 12 months. We can speculate that during normal aging, there is a gradual increase of the barrage of excitatory and inhibitory synaptic events received by CA1 pyramidal cells.
More time points should be investigated to test this hypothesis. Several non-excluding mechanisms can explain such a rise, including an increase in the number of synapses, more active presynaptic cells, and a greater release probability from the presynaptic terminals.
This result is consistent with that reported in the prefrontal cortex during aging (using later time points) for animals preserving their cognitive performances (Bories et al., 2013). Interestingly, in Tau mice on water, both glutamatergic and GABAergic drives were increased at 8 and 12 months as compared to their Wild type counterparts. In keeping with our results, most studies using Tau or Aß models report an increase in the excitatory drive as compared to Wild Type (Crimins et al., 2011; Dalby et al., 2014; Ovsepian et al., 2017), but see Rocher et al. (2008).
Less data is available for the GABAergic drive. In the cortex of a different Tau model, there was no difference in sIPSC frequency at 9 months as compared to Wild Type (Crimins et al., 2011). The discrepancy with our results may stem from the type of mutant used and the brain region selected. Together, our results raise the intriguing possibility that the Tau mutation accelerates the aging process as assessed with the glutamatergic and GABAergic drives received by CA1 pyramidal cells.
Early life exposure to caffeine changed the apparent co-variance relationship between the two age and Tau independent variables. At 8 months, both GABAergic and glutamatergic drives were decreased in Tau caffeine mice as compared to Tau water mice. However, at 12 months, although the excitatory drive was decreased as compared to that measured in Tau water mice, the inhibitory drive was increased.
Hence, contrary to our hypothesis, caffeine exposure does not accelerate the increase in synaptic barrage received by CA1 pyramidal cells that occurs in water exposed animal. We propose that the three variables tau-caffeine-age interact in non-linear fashion, thus constituting a complex system (the global behavior cannot be predicted from the observation of its independent components). This exemplifies the difficulty in interpreting the in vitro data.
However, our data emphasize that caffeine exposure disrupts the effect of the pathogenic process characteristic of the Tau phenotype in CA1 pyramidal cells. If there is a correlation between cognitive deficits and electrophysiological alterations at the circuit level, more parameters need to be measured (e.g., ion channels or metabolism). The possibility also exists that the CA1 region is not the most appropriate to establish such a correlation. In vivo recordings may also provide a different entry point into the underlying mechanisms, in particular measuring the properties of brain rhythms (theta, gamma) and sleep patterns.
reference link: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6797851/
More information: Zachary P. Christensen et al, Caffeine exposure in utero is associated with structural brain alterations and deleterious neurocognitive outcomes in 9–10 year old children, Neuropharmacology (2021). DOI: 10.1016/j.neuropharm.2021.108479