Male-specific deficits in natural reward learning in a mouse model of neurodevelopmental disorders.

Neurodevelopmental disorders, including autism spectrum disorders, are highly male biased, but the underpinnings of this are unknown. Striatal dysfunction has been strongly implicated in the pathophysiology of neurodevelopmental disorders, raising the question of whether there are sex differences in how the striatum is impacted by genetic risk factors linked to neurodevelopmental disorders. Here we report male-specific deficits in striatal function important to reward learning in a mouse model of 16p11.2 hemideletion, a genetic mutation that is strongly associated with the risk of neurodevelopmental disorders, particularly autism and attention-deficit hyperactivity disorder. We find that male, but not female, 16p11.2 deletion animals show impairments in reward-directed learning and maintaining motivation to work for rewards. Male, but not female, deletion animals overexpress mRNA for dopamine receptor 2 and adenosine receptor 2a in the striatum, markers of medium spiny neurons signaling via the indirect pathway, associated with behavioral inhibition. Both sexes show a 50% reduction of mRNA levels of the genes located within the 16p11.2 region in the striatum, including the kinase extracellular-signal related kinase 1 (ERK1). However, hemideletion males show increased activation in the striatum for ERK1, both at baseline and in response to sucrose, a signaling change associated with decreased striatal plasticity. This increase in ERK1 phosphorylation is coupled with a decrease in the abundance of the ERK phosphatase striatum-enriched protein-tyrosine phosphatase in hemideletion males. In contrast, females do not show activation of ERK1 in response to sucrose, but notably hemideletion females show elevated protein levels for ERK1 as well as the related kinase ERK2 over what would be predicted by mRNA levels. These data indicate profound sex differences in the impact of a genetic lesion linked with neurodevelopmental disorders, including mechanisms of male-specific vulnerability and female-specific resilience impacting intracellular signaling in the brain.

Developmental sensitivity of associative learning to cholesterol synthesis inhibitors.

Patients with Smith-Lemli-Opitz syndrome, a genetic disorder associated with severe mental retardation, are unable to convert 7-dehydrocholesterol to cholesterol. Treatment of rats with agents that block cholesterol synthesis produces a sterol profile reminiscent of Smith-Lemli-Opitz patients i.e., low levels of cholesterol accompanied by the appearance of its immediate precursor 7-dehydrocholesterol. In previous work, chronic inhibition of cholesterol synthesis in just-weaned rats impaired acquisition of the classically conditioned eyeblink response. The present study had two primary goals--(1) to determine whether the learning impairment depended on the age in which treatment was initiated; and (2) to determine whether the deficit was associative or due to performance factors. Consistent with earlier work, acquisition of the eyeblink conditioned response was impaired when the 30-day treatment was initiated on postnatal day (PND) 21. Reactivity to acoustic stimuli and to eyelid stimulation were normal, suggesting that the learning impairment was associative in nature. The learning impairment was transitory; acquisition was normal when evaluated 30 days after the cessation of treatment. When treatment was initiated 30 days after weaning (PND 51), acquisition of the eyeblink response was normal. However, brain sterols of young adult rats were less affected than those of just-weaned rats. Thus, there is a developmental sensitivity to cholesterol synthesis blocking agents both in terms of their effects on brain sterols and new motor learning.

Blocking cholesterol synthesis impairs acquisition of the classically conditioned eyeblink response.

Smith-Lemli-Opitz (SLO) syndrome is a congenital disorder characterized by severe mental retardation. Patients with SLO lack 7-dehydrocholesterol (7 dH) reductase, which catalyzes the last step of cholesterol synthesis. Administration of an agent that blocks 7 dH cholesterol reductase, BM 15.766 (BM), leads to a biochemical profile which resembles that of SLO patients, i.e., lower plasma, liver, and brain cholesterol levels accompanied by the appearance of the precursors 7 dH and 8 dH cholesterol. In this article we address the functional consequences of chronic BM treatment on new motor learning by assessing acquisition of the classically conditioned eyeblink response. Just-weaned rats were fed BM by gavage for four months, with half of these rats given exogenous cholesterol during the last two months of BM treatment. Acquisition of the eyeblink response was impaired in BM-treated rats. Impaired acquisition of the eyeblink response was not accompanied by alterations in responsiveness to either the conditioned or unconditioned stimulus. Exogenous cholesterol, a clinically relevant countertreatment, failed to correct for the learning impairment produced by BM treatment. Chronic treatment with a cholesterol synthesis-blocking agent impaired associative learning in just-weaned rats.

Relationship between abnormal cholesterol synthesis and retarded learning in rats.

We examined the relationship between brain sterol composition and associative learning (classical conditioning of the eyeblink response) in newly weaned rats fed BM 15.766 (BM) for 4 months. This compound inhibits 7-dehydrocholesterol-delta7-reductase, which catalyzes the conversion of 7-dehydrocholesterol to cholesterol, the last step in the synthetic pathway. As countertreatment, half of the BM-treated rats were fed 2% cholesterol during the last 2 months. With BM, cholesterol concentrations declined 91% in plasma, but with cholesterol feeding, the levels increased 50% compared with baseline values. 7-Dehydrocholesterol, which was not detected at baseline, increased to 55% of plasma sterols with BM but decreased to 5% of total plasma sterols when cholesterol was added. With BM, brain cholesterol levels decreased 60% and did not increase after cholesterol was added. However, 7-dehydrocholesterol, which comprised 39% of brain sterols with BM, decreased to 31% (P < .05) when cholesterol was fed. Hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase activity in the liver increased 2.2-fold with BM and declined 95% after adding cholesterol, but did not change in the brain. BM treatment for 4 months prevented learning of the conditioned eyeblink response as compared with controls. In contrast, BM-treated rats supplemented with cholesterol acquired the conditioned eyeblink response. Chronic inhibition of 7-dehydrocholesterol-delta7-reductase reduced cholesterol and increased 7-dehydrocholesterol levels in plasma and brain, and was associated with impaired learning. Cholesterol feeding corrected plasma and hepatic sterol levels and reduced brain 7-dehydrocholesterol concentrations to reestablish normal learning.

Intrinsic GUS-like activities in seed plants.

Fifty-two plant species, covering some Gymnosperms and all the key groups of Angiosperms, were chosen for surveying their intrinsic beta-glucuronidase-like activities. Histochemical (overnight incubation) and qualitative fluorometric (24 h incubation) assays indicated that, with few exceptions, such activities were detected in certain part(s) of the fruit walls, seed coats, endosperms or, especially, the embryos of the tested plants. Most of such activities in the excised immature embryos of soybean and string bean disappeared after one to a few days' in vitro culturing. Such activities in the intact mature seeds of these two species diminished also during germination process. The vegetative organs of seedlings/mature plants usually lack such activities. The enzyme(s) responsible for such activities was antigenically dissimilar to E. coli beta-glucuronidase.