The behavioral effects of gestational and lactational benzo[a]pyrene exposure vary by sex and genotype in mice with differences at the Ahr and Cyp1a2 loci

Benzo[a]pyrene (BaP) is a polycyclic aromatic hydrocarbon (PAH) and known carcinogen in the Top 10 on the United States' list of priority pollutants. Humans are exposed through a variety of sources including tobacco smoke, grilled foods and fossil fuel combustion. Recent studies of children exposed to higher levels of PAHs during pregnancy and early life have identified numerous adverse effects on the brain and behavior that persist into school age and adolescence. Our studies were designed to look for genotype and sex differences in susceptibility to gestational and lactational exposure to BaP using a mouse model with allelic differences in the aryl hydrocarbon receptor and the xenobiotic metabolizing enzyme CYP1A2. Pregnant dams were exposed to 10 mg/kg/day of BaP in corn oil-soaked cereal or the corn oil vehicle alone from gestational day 10 until weaning at postnatal day 25. Neurobehavioral testing began at P60 using one male and one female per litter. We found main effects of sex, genotype and treatment as well as significant gene x treatment and sex x treatment interactions. BaP-treated female mice had shorter latencies to fall in the Rotarod test. High-affinity AhrbCyp1a2(-/-) mice had greater impairments in Morris water maze. Interestingly, poor-affinity AhrdCyp1a2(-/-) mice also had deficits in spatial learning and memory regardless of treatment. We believe our findings provide future directions in identifying human populations at highest risk of early life BaP exposure, because our model mimics known human variation in our genes of interest. Our studies also highlight the value of testing both males and females in all neurobehavioral studies.

Benson’s Disease or Posterior Cortical Atrophy, Revisited

Background: D. Frank Benson and colleagues first described the clinical and neuropathological features of posterior cortical atrophy (PCA) from patients in the UCLA Neurobehavior Program. Objective: We reviewed the Program’s subsequent clinical experience with PCA, and its potential for clarifying this relatively rare syndrome in comparison to the accumulated literature on PCA. Methods: Using the original criteria derived from this clinic, 65 patients with neuroimaging-supported PCA were diagnosed between 1995 and 2020. Results: On presentation, most have visual localization complaints and related visuospatial symptoms, but nearly half had memory complaints followed by symptoms of depression. Neurobehavioral testing showed predominant difficulty with visuospatial constructions, Gerstmann’s syndrome, and Balint’s syndrome, but also impaired memory and naming. On retrospective application of the current Consensus Criteria for PCA, 59 (91%) met PCA criteria with a modification allowing for “significantly greater visuospatial over memory and naming deficits.” There were 37 deaths (56.9%) with the median overall survival of 10.3 years (95% CI: 9.6–13.6 years), consistent with a slow neurodegenerative disorder in most patients. Conclusion: Together, these findings recommend modifying the PCA criteria for “relatively spared” memory, language, and behavior to include secondary memory and naming difficulty and depression, with increased emphasis on the presence of Gerstmann’s and Balint’s syndromes.

075 Systematic Individual Differences in Vulnerability to Sleep Inertia

Introduction Sleep inertia (SI), the transient grogginess and disorientation that occurs upon awakening, may be particularly problematic in on-call operations that require safety-sensitive and time-critical action, such as healthcare and emergency response. The magnitude of SI is determined by multiple factors, including sleep history, and anecdotal evidence suggests individuals may differ considerably in susceptibility to impairment from SI. Methods As part of a larger study investigating individual differences in neurobehavioral impairment, N=21 healthy adults (aged 21–38y; 9 females) completed three laboratory-based neurobehavioral testing sessions, each preceded by baseline sleep. Baseline sleep and nightly at-home sleep opportunities during the week prior were either 12h (extended) or 6h (restricted); two sessions involved extension and one involved restriction in randomized, counterbalanced order. Baseline sleep opportunities ended at 10:00, whereupon subjects completed a 60min neurobehavioral test battery, which began with the Karolinska Sleepiness Scale (KSS). The test battery was repeated every 2h throughout the testing sessions. Results A nonlinear mixed-effects regression, controlling for prior sleep restriction/extension and session number, was used to estimate the subjective magnitude of SI immediately upon awakening from baseline sleep, as measured by KSS scores, and the exponential dissipation rate of the effect relative to KSS scores later in the day (12:00-20:00). Following prior sleep extension, SI was associated with a 1.82±0.59 KSS score increase (p=0.006), which subsequently dissipated from a level of 4.80±0.65 to 2.98±0.29 (p=<0.001) later in the day. Following prior sleep restriction, SI was associated with a 1.58±0.58 KSS score increase (p=0.014), which subsequently dissipated from a level of 5.39±0.63 to 3.80±0.30 later in the day. SI took ~45min to dissipate to a negligible level based on a time constant estimate of 23.6±15.8min. Importantly, there were substantial, systematic individual differences in the magnitude of SI, with a between-subjects standard deviation of 1.15±0.42 points on the KSS (ICC=0.51, F=19.5, p<0.001). Conclusion We observed sizeable, systematic individual differences in subjective sleepiness due to sleep inertia. To what degree these individual differences predict objective performance deficits remains to be investigated. Support (if any) NASA grant NAG9-1161 and CDMRP grant W81XWH-20-1-0442

Earlier preterm birth is associated with a worse neurocognitive outcome in a rabbit model

Background Preterm birth (PTB) and particularly late preterm PTB has become a research focus for obstetricians, perinatologists, neonatologists, pediatricians and policy makers alike. Translational models are useful tools to expedite and guide clinical but presently no model exists that contextualizes the late PTB scenario. Herein we aimed to develop a rabbit model that echo’s the clinical neurocognitive phenotypes of early and late PTB. Methods Time mated rabbit does underwent caesarean delivery at a postconceptional age (PCA) of either 28 (n = 6), 29 (n = 5), 30 (n = 4) or 31 (n = 4) days, term = 31 d. Newborn rabbits were mixed and randomly allocated to be raised by cross fostering and underwent short term neurobehavioral testing on corrected post-natal day 1. Open field (OFT), spontaneous alteration (TMT) and novel object recognition (NORT) tests were subsequently performed at 4 and 8 weeks of age. Results PTB was associated with a significant gradient of short-term mortality and morbidity inversely related to the PCA. On postnatal day 1 PTB was associated with a significant sensory deficit in all groups but a clear motor insult was only noted in the PCA 29d and PCA 28d groups. Furthermore, PCA 29d and PCA 28d rabbits had a persistent neurobehavioral deficit with less exploration and hyperanxious state in the OFT, less alternation in TMT and lower discriminatory index in the NORT. While PCA 30d rabbits had some anxiety behavior and lower spontaneous alteration at 4 weeks, however at 8 weeks only mild anxiety driven behavior was observed in some of these rabbits. Conclusions In this rabbit model, delivery at PCA 29d and PCA 28d mimics the clinical phenotype of early PTB while delivery at PCA 30d resembles that of late PTB. This could serve as a model to investigate perinatal insults during the early and late preterm period.

The absence of the CD163 receptor has distinct temporal influences on intracerebral hemorrhage outcomes

Hemoglobin (Hb) toxicity precipitates secondary brain damage following intracerebral hemorrhage (ICH). CD163 is an anti-inflammatory Hb scavenger receptor and CD163-positive macrophages/microglia locally accumulate post-bleed, yet no studies have investigated the role of CD163 after ICH. ICH was induced in wildtype and CD163−/− mice and various anatomical and functional outcomes were assessed. At 3 d, CD163−/− mice have 43.4 ± 5.0% (p = 0.0002) and 34.8 ± 3.4% (p = 0.0003) less hematoma volume and tissue injury, respectively. Whereas, at 10 d, CD163−/− mice have 49.2 ± 15.0% larger lesions (p = 0.0385). An inflection point was identified, where CD163−/− mice perform better on neurobehavioral testing and have less mortality before 4 d, but increased mortality and worse function after 4 d (p = 0.0389). At 3 d, CD163−/− mice have less Hb, iron, and blood–brain barrier dysfunction, increased astrogliosis and neovascularization, and no change in heme oxygenase 1 (HO1) expression. At 10 d, CD163−/− mice have increased iron and VEGF immunoreactivity, but no significant change in HO1 or astrogliosis. These novel findings reveal that CD163 deficiency has distinct temporal influences following ICH, with early beneficial properties but delayed injurious effects. While it is unclear why CD163 deficiency is initially beneficial, the late injurious effects are consistent with the key anti-inflammatory role of CD163 in the recovery phase of tissue damage.

Abstract 151: CD163 Has Distinct Temporal Influences on Intracerebral Hemorrhage Outcomes

Extracorpuscular hemoglobin (Hb)-induced toxicity following intracerebral hemorrhage (ICH) precipitates secondary brain damage and poor outcomes. CD163 is the Hb scavenger receptor with potent anti-inflammatory effects and CD163-positive macrophages/microglia accumulate in the brain with time post-ICH, yet no studies have investigated the role of CD163 after ICH. ICH was induced in wildtype and CD163 -/- mice and various anatomical and functional outcomes were temporally assessed by histology and neurobehavioral testing. Acutely, CD163 -/- mice have 33.2±4.5% (p<0.0001), 43.4±5.0% (p=0.0002), and 34.8±3.4% (p=0.0003) less lesion volume, hematoma volume, and tissue injury, respectively. Whereas, at 10d, CD163 -/- mice have 49.2±15.0% larger lesion volumes (p=0.0385). Temporal data inspection revealed an inflection point at 4d, where CD163 -/- mice perform significantly better on neurobehavioral testing and have less mortality before 4d, but increased mortality and worse function after 4d (p<0.05). Histology at 72h shows that CD163 -/- mice have significantly less Hb, iron, and blood-brain barrier dysfunction, increased astrogliosis and cortical neovascularization, and no change in HO1 expression. At 10d, CD163 -/- mice have increased iron and hematomal VEGF immunoreactivity, no change in HO1 expression, and decreased astrogliosis. These novel findings reveal that CD163 deficiency has distinct temporal influences on ICH outcomes, with early beneficial properties but delayed injurious effects. While it is unclear why CD163 deficiency is initially beneficial, the late injurious effects of absent CD163 are consistent with the key anti-inflammatory role of the receptor in the recovery phase of tissue damage. CD163 may represent a key targetable immunomodulatory receptor after ICH. Funding: This work was supported by NIH grants F31NS086441 (JLL) and R01NS046400 (SD).

Neurobehavioral testing in subarachnoid hemorrhage: A review of methods and current findings in rodents

The most important aspect of a preclinical study seeking to develop a novel therapy for neurological diseases is whether the therapy produces any clinically relevant functional recovery. For this purpose, neurobehavioral tests are commonly used to evaluate the neuroprotective efficacy of treatments in a wide array of cerebrovascular diseases and neurotrauma. Their use, however, has been limited in experimental subarachnoid hemorrhage studies. After several randomized, double-blinded, controlled clinical trials repeatedly failed to produce a benefit in functional outcome despite some improvement in angiographic vasospasm, more rigorous methods of neurobehavioral testing became critical to provide a more comprehensive evaluation of the functional efficacy of proposed treatments. While several subarachnoid hemorrhage studies have incorporated an array of neurobehavioral assays, a standardized methodology has not been agreed upon. Here, we review neurobehavioral tests for rodents and their potential application to subarachnoid hemorrhage studies. Developing a standardized neurobehavioral testing regimen in rodent studies of subarachnoid hemorrhage would allow for better comparison of results between laboratories and a better prediction of what interventions would produce functional benefits in humans.