Dynamin 2 is required for GPVI signaling and platelet hemostatic function in mice.

Receptor-mediated endocytosis, which contributes to a wide range of cellular functions, including receptor signaling, cell adhesion, and migration, requires endocytic vesicle release by the large GTPase dynamin 2. Here, the role of dynamin 2 was investigated in platelet hemostatic function using both pharmacological and genetic approaches. Dnm2fl/fl Pf4-Cre (Dnm2Plt - / -) mice specifically lacking dynamin 2 within the platelet lineage developed severe thrombocytopenia and bleeding diathesis and Dnm2Plt - / - platelets adhered poorly to collagen under arterial shear rates. Signaling via the collagen receptor GPVI was impaired in platelets treated with the dynamin GTPase inhibitor dynasore, as evidenced by poor protein tyrosine phosphorylation, including that of the proximal tyrosine kinase Lyn on its activating tyrosine 396 residue. Platelet stimulation via GPVI resulted in a slight decrease in GPVI, which was maintained by dynasore treatment. Dynasore-treated platelets had attenuated function when stimulated via GPVI, as evidenced by reduced GPIbα downregulation, α-granule release, integrin αIIbß3 activation, and spreading onto immobilized fibrinogen. By contrast, responses to the G-protein coupled receptor agonist thrombin were minimally affected by dynasore treatment. GPVI expression was severely reduced in Dnm2Plt-/- platelets, which were dysfunctional in response to stimulation via GPVI, and to a lesser extent to thrombin. Dnm2Plt-/- platelets lacked fibrinogen in their α-granules, but retained von Willebrand factor. Taken together, the data show that dynamin 2 plays a proximal role in signaling via the collagen receptor GPVI and is required for fibrinogen uptake and normal platelet hemostatic function.

The Adult Murine Intestine is Dependent on Constitutive Laminin-γ1 Synthesis.

Laminin-γ1 is required for early embryonic development; however, the need for laminin-γ1 synthesis in adulthood is unknown. A global and inducible mouse model of laminin-γ1 deficiency was generated to address this question. Genetic ablation of the Lamc1 gene in adult mice was rapidly lethal. Despite global Lamc1 gene deletion in tamoxifen-induced mutant mice, there was minimal change in total cardiac, pulmonary, hepatic or renal laminin protein. In contrast, laminin-γ1 was significantly depleted in the small intestines, which showed crypt hyperplasia and dissociation of villous epithelium from adjacent mesenchyme. We conclude that the physiologic requirement for laminin-γ1 synthesis in adult mice is dependent on a tissue-specific basal rate of laminin-γ1 turnover that results in rapid depletion of laminin-γ1 in the intestine.

Impact of cannabis use on prefrontal and parietal cortex gyrification and surface area in adolescents and emerging adults.

BACKGROUND: Regions undergoing maturation with CB1 receptors may be at increased risk for cannabis-induced alterations. Here, we examine the relationships between cannabis use and prefrontal (PFC) and inferior parietal gyrification and surface area (SA) in youth. METHODS: Participants included 33 cannabis users and 35 controls (ages 18-25). Exclusions included co-morbid psychiatric/neurologic disorders and heavy other drug use. Multiple regressions and Pearson r correlations examined the effects of cannabis use on gyrification, SA and cognition. RESULTS: Cannabis use was associated with decreased gyrification in: ventral-medial PFC (RH: [FDR corrected p=.02], LH: [FDR corrected p=.02]); medial PFC (RH: [FDR corrected p=.02], LH: [FDR corrected p=.02]); and frontal poles (RH: [FDR corrected p=.02], LH: [FDR corrected p=.02]). No differences were observed in bilateral hemispheres, PFC, dorsolateral, ventrolateral, or inferior parietal ROIs. Cannabis use was associated with marginally decreased SA in left: medial PFC [FDR corrected p=.09], and ventral lateral PFC: [FDR corrected p=.09]. In cannabis users, increased gyrification was associated with improved working-memory performance in right medial (p=.003), ventral-medial (p=.03), and frontal pole ROIs (p=.007). CONCLUSIONS: Cannabis use was associated with reduced gyrification in PFC regions implicated in self-referential thought and social cognition. Results suggest that these gyrification characteristics may have cognitive implications.

Poorer frontolimbic white matter integrity is associated with chronic cannabis use, FAAH genotype, and increased depressive and apathy symptoms in adolescents and young adults.

BACKGROUND: The heaviest period of cannabis use coincides with ongoing white matter (WM) maturation. Further, cannabis-related changes may be moderated by FAAH genotype (rs324420). We examined the association between cannabis use and FAAH genotype on frontolimbic WM integrity in adolescents and emerging adults. We then tested whether observed WM abnormalities were linked with depressive or apathy symptoms. METHODS: Participants included 37 cannabis users and 37 healthy controls (33 female; ages 18-25). Multiple regressions examined the independent and interactive effects of variables on WM integrity. RESULTS: Regular cannabis users demonstrated reduced WM integrity in the bilateral uncinate fasciculus (UNC) (MD, right: p = .009 and left: p = .009; FA, right: p = .04 and left: p = .03) and forceps minor (fMinor) (MD, p = .03) compared to healthy controls. Marginally reduced WM integrity in the cannabis users was found in the left anterior thalamic radiation (ATR) (FA, p = .08). Cannabis group ∗ FAAH genotype interaction predicted WM integrity in bilateral ATR (FA, right: p = .05 and left: p = .001) and fMinor (FA, p = .02). In cannabis users, poorer WM integrity was correlated with increased symptoms of depression and apathy in bilateral ATR and UNC. CONCLUSIONS: Consistent with prior findings, cannabis use was associated with reduced frontolimbic WM integrity. WM integrity was also moderated by FAAH genotype, in that cannabis-using FAAH C/C carriers and A carrying controls had reduced WM integrity compared to control C/C carriers. Observed frontolimbic white matter abnormalities were linked with increased depressive and apathy symptoms in the cannabis users.

Effects of marijuana use on prefrontal and parietal volumes and cognition in emerging adults.

RATIONALE: Chronic marijuana (MJ) use among adolescents has been associated with structural and functional abnormalities, particularly in developing regions responsible for higher order cognition. OBJECTIVES: This study investigated prefrontal (PFC) and parietal volumes and executive function in emerging adult MJ users and explored potential gender differences. METHODS: Participants (ages 18-25) were 27 MJ users and 32 controls without neurologic or psychiatric disorders or heavy other drug use. A series of multiple regressions examined whether group status, past year MJ use, and their interactions with gender predicted ROI volumes. Post hoc analyses consisted of brain-behavior correlations between volumes and cognitive variables and Fisher's z tests to assess group differences. RESULTS: MJ users demonstrated significantly smaller medial orbitofrontal (mOFC; p = 0.004, FDR p = 0.024) and inferior parietal volumes (p = 0.04, FDR p = 0.12); follow-up regressions found that increased past year MJ use did not significantly dose-dependently predict smaller mOFC volume in a sub-sample of individuals with at least one past year MJ use. There were no significant gender interactions. There was a significant brain-behavior difference by group, such that smaller mOFC volumes were associated with poorer complex attention for MJ users (p < 0.05). CONCLUSIONS: Smaller mOFC volumes among MJ users suggest disruption of typical neurodevelopmental processes associated with regular MJ use for both genders. These results highlight the need for longitudinal, multi-modal imaging studies providing clearer information on timing of neurodevelopmental processes and neurocognitive impacts of youth MJ initiation.

Learning redundant motor tasks with and without overlapping dimensions: facilitation and interference effects.

Prior learning of a motor skill creates motor memories that can facilitate or interfere with learning of new, but related, motor skills. One hypothesis of motor learning posits that for a sensorimotor task with redundant degrees of freedom, the nervous system learns the geometric structure of the task and improves performance by selectively operating within that task space. We tested this hypothesis by examining if transfer of learning between two tasks depends on shared dimensionality between their respective task spaces. Human participants wore a data glove and learned to manipulate a computer cursor by moving their fingers. Separate groups of participants learned two tasks: a prior task that was unique to each group and a criterion task that was common to all groups. We manipulated the mapping between finger motions and cursor positions in the prior task to define task spaces that either shared or did not share the task space dimensions (x-y axes) of the criterion task. We found that if the prior task shared task dimensions with the criterion task, there was an initial facilitation in criterion task performance. However, if the prior task did not share task dimensions with the criterion task, there was prolonged interference in learning the criterion task due to participants finding inefficient task solutions. These results show that the nervous system learns the task space through practice, and that the degree of shared task space dimensionality influences the extent to which prior experience transfers to subsequent learning of related motor skills.

The effect of movement rate and complexity on functional magnetic resonance signal change during pedaling.

We used functional magnetic resonance imaging (fMRI) to record human brain activity during slow (30 RPM), fast (60 RPM), passive (30 RPM), and variable rate pedaling. Ten healthy adults participated. After identifying regions of interest, the intensity and volume of brain activation in each region was calculated and compared across conditions (p < .05). Results showed that the primary sensory and motor cortices (S1, M1), supplementary motor area (SMA), and cerebellum (Cb) were active during pedaling. The intensity of activity in these areas increased with increasing pedaling rate and complexity. The Cb was the only brain region that showed significantly lower activity during passive as compared with active pedaling. We conclude that M1, S1, SMA, and Cb have a role in modifying continuous, bilateral, multijoint lower extremity movements. Much of this brain activity may be driven by sensory signals from the moving limbs.

Pedaling alters the excitability and modulation of vastus medialis H-reflexes after stroke.

OBJECTIVE: Individuals post-stroke display abnormal Group Ia reflex excitability. Pedaling has been shown to reduce Group Ia reflexes and to normalize the relationship between EMG and reflex amplitude in the paretic soleus (SO). The purpose of this study was to determine whether these changes extend to the paretic quadriceps. METHODS: H-reflexes were used to examine Group Ia reflex excitability of the vastus medialis (VM). H-reflexes were elicited in paretic (n=13) and neurologically intact (n=13) individuals at 11 positions in the pedaling cycle and during static knee extension at comparable limb positions and levels of VM EMG. RESULTS: VM H-reflexes were abnormally elevated in the paretic limb of stroke survivors. During static muscle activation, H-reflex amplitude did not change with the level of background VM activity. Pedaling reduced the amplitude of paretic VM H-reflexes and restored the normal relationship between VM EMG and H-reflex amplitude. CONCLUSIONS: Pedaling-induced changes in Group Ia reflex excitability that have been reported for the paretic SO are evident in the paretic VM. Pedaling may have a generalized effect on lower extremity Group Ia reflexes post-stroke. SIGNIFICANCE: Pedaling may be therapeutic for reducing Group Ia reflexes after stroke.

A novel technique for examining human brain activity associated with pedaling using fMRI.

Advances in neural imaging technologies, such as functional magnetic resonance imaging (fMRI), have made it possible to obtain images of human brain activity during motor tasks. However, technical challenges have made it difficult to image the brain during multijoint lower limb movements like those involved in locomotion. We developed an MR compatible pedaling device and recorded human brain activity associated with rhythmic, alternating flexion and extension of the lower extremities. Ten volunteers pedaled at 30 RPM while recording fMRI signals in a GE 3T short bore MR scanner. We utilized a block design consisting of 3 runs of pedaling, each lasting 4 min. In a single run, subjects pedaled for 30 s and then rested for 30 s. This sequence was repeated 4 times. Conventional fMRI processing techniques, that correlate the entire BOLD signal with standard model, did not extract physiologically meaningful signal, likely due to magnetic field distortion caused by leg movement. Hence, we examined only the portion of the blood-oxygen-level dependent (BOLD) signal during movement-free periods. This technique takes advantage of the delayed nature of the BOLD signal and fits the falling portion of the signal after movement has stopped with a standard model. Using this approach, we observed physiologically plausible brain activity patterns associated with pedaling in the primary and secondary sensory and motor cortices and the cerebellum. To our knowledge, this is the first time that human brain activity associated with pedaling has been recorded with fMRI. This technique may be useful for advancing our understanding of supraspinal control of locomotor-like movements in health and disease.

Soleus H-reflex excitability during pedaling post-stroke.

A major contributor to impaired locomotion post-stroke is abnormal phasing of paretic muscle activity, but the mechanisms remain unclear. Previous studies have shown that, in the paretic limb of people post-stroke, Group Ia reflexes are abnormally elevated and fail to decrease in amplitude during locomotion. Hence, we hypothesized that inappropriate muscle phasing may be associated with enhanced transmission in the monosynaptic Group Ia afferent pathway. Soleus (SO) H-reflexes were used to examine transmission in the Group Ia afferent pathway to SO motor neurons during pedaling, a locomotor task in which abnormal muscle phasing is evident. Our hypothesis predicted that H-reflexes would be elevated during the flexion phase of pedaling where inappropriate SO activity occurs. H-reflexes were elicited in paretic (n = 13) and neurologically intact (NI, n = 26) individuals at 11 different positions in the pedaling cycle and during tonic plantar flexion at comparable limb positions and levels of SO EMG. In both groups, SO H-reflexes were smaller during pedaling as compared to matched tonic plantar flexion. In the NI group, but not the paretic group, SO H-reflex amplitude was significantly modulated across the pedaling cycle. H-reflexes were large during extension and small during flexion. Reduced H-reflex modulation post-stroke was associated with the level of neuromuscular impairment as indicated by Fugl-Meyer score. However, regardless of impairment level, stroke subjects displayed H-reflex suppression during the flexion phase of pedaling. After correcting for the level of background muscle activity, H-reflexes were found to be larger in paretic as compared to NI individuals, regardless of the phase of the pedaling cycle. We conclude that Group Ia afferent transmission is enhanced in the paretic SO of people post-stroke as compared to NI individuals. However, contrary to our hypothesis, enhanced transmission in the Group Ia monosynaptic spinal pathway is not specifically associated with extraneous extensor muscle activity during the flexion phase of pedaling and is unlikely to account for abnormal locomotor muscle phasing post-stroke. This result is important because it suggests that, despite the presence of hyperactive monosynaptic reflexes post-stroke, this impairment may not make an important contribution to abnormal locomotor muscle activity.