Rebecca A. Johnson

BDNF is necessary and sufficient for spinal respiratory plasticity following intermittent hypoxia

Intermittent hypoxia causes a form of serotonin-dependent synaptic plasticity in the spinal cord known as phrenic long-term facilitation (pLTF). Here we show that increased synthesis of brain-derived neurotrophic factor (BDNF) in the spinal cord is necessary and sufficient for pLTF in adult rats. We found that intermittent hypoxia elicited serotonin-dependent increases in BDNF synthesis in ventral spinal segments containing the phrenic nucleus, and the magnitude of these BDNF increases correlated with pLTF magnitude. We used RNA interference (RNAi) to interfere with BDNF expression, and tyrosine kinase receptor inhibition to block BDNF signaling. These disruptions blocked pLTF, whereas intrathecal injection of BDNF elicited an effect similar to pLTF. Our findings demonstrate new roles and regulatory mechanisms for BDNF in the spinal cord and suggest new therapeutic strategies for treating breathing disorders such as respiratory insufficiency after spinal injury. These experiments also illustrate the potential use of RNAi to investigate functional consequences of gene expression in the mammalian nervous system in vivo.

Hippocampal brain-derived neurotrophic factor but not neurotrophin-3 increases more in mice selected for increased voluntary wheel running.

Voluntary wheel running in rats increases hippocampal brain-derived neurotrophic factor (BDNF) expression, a neurochemical important for neuronal survival, differentiation, connectivity and synaptic plasticity. Here, we report the effects of wheel running on BDNF and neurotrophin-3 (NT-3) protein levels in normal control mice, and in mice selectively bred (25 generations) for increased voluntary wheel running. We hypothesized that increased voluntary wheel running in selected (S) mice would increase CNS BDNF and NT-3 protein levels more than in control (C) mice. Baseline hippocampal BDNF levels (mice housed without running wheels) were similar in S and C mice. Following seven nights of running, hippocampal BDNF increased significantly more in S versus C mice, and levels were correlated with distance run (considering C and S mice together). Spinal and cerebellar BDNF and hippocampal NT-3 levels were not significantly affected by wheel running in any group, but there was a small, positive correlation between spinal C3-C6 BDNF levels and distance run (considering C and S mice together). This is the first study to demonstrate that mice which choose to run more have greater elevations in hippocampal BDNF, suggesting enhanced potential for exercise-induced hippocampal neuroplasticity.

Exercise-induced changes in hippocampal brain-derived neurotrophic factor and neurotrophin-3: effects of rat strain.

We tested the hypothesis that exercise-induced changes in hippocampal brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) differ among rat strains exhibiting a range of voluntary wheel running activity. Four strains (Sprague-Dawley, Brown Norway, Dark Agouti and PVG) were given access to running wheels (1 or 7 nights). Over 7 nights, the average distance run per night was higher in PVG versus other strains, and higher in Brown Norway versus Sprague-Dawley rats. Hippocampal BDNF concentrations in sedentary rats were higher in PVG versus Sprague-Dawley rats. When data from all strains were combined, BDNF levels increased with 7 nights of wheel running and were positively correlated to the previous night distance run. Sedentary hippocampal NT-3 levels were not different between rat strains, but decreased with 7 nights of wheel access; NT-3 was negatively correlated with previous night distance run. There were no differences between strains in the correlation between distance run and BDNF or NT-3 levels. Although exercise decreases hippocampal NT-3, strain does not alter NT-3 levels. In contrast, BDNF levels increase with exercise and basal levels differ between strains, possibly due to strain differences in spontaneous activity.

7,8-dihydroxyflavone exhibits therapeutic efficacy in a mouse model of Rett syndrome

Rett syndrome (RTT), caused by mutations in the methyl-CpG binding protein 2 gene (MECP2), is a debilitating autism spectrum developmental disorder predominantly affecting females. Mecp2 mutant mice have reduced levels of brain-derived neurotrophic factor (BDNF) in the brain; conditional deletion and overexpression of BDNF in the brain accelerates and slows, respectively, disease progression in Mecp2 mutant mice. Thus we tested the hypothesis that 7,8-dihydroxyflavone (7,8-DHF), a small molecule reported to activate the high affinity BDNF receptor (TrkB) in the CNS, would attenuate disease progression in Mecp2 mutant mice. Following weaning, 7,8-DHF was administered in drinking water throughout life. Treated mutant mice lived significantly longer compared with untreated mutant littermates (80 ± 4 and 66 ± 2 days, respectively). 7,8-DHF delayed body weight loss, increased neuronal nuclei size and enhanced voluntary locomotor (running wheel) distance in Mecp2 mutant mice. In addition, administration of 7,8-DHF partially improved breathing pattern irregularities and returned tidal volumes to near wild-type levels. Thus although the specific mechanisms are not completely known, 7,8-DHF appears to reduce disease symptoms in Mecp2 mutant mice and may have potential as a therapeutic treatment for RTT patients.

Chronic cervical spinal sensory denervation reveals ineffective spinal pathways to phrenic motoneurons in the rat

We hypothesized that pretreatment with chronic cervical dorsal rhizotomy (CDR; C(3)-C(6)) would reveal ineffective crossed spinal pathways to phrenic motoneurons. Anesthetized CDR (1 week post-rhizotomy) and control rats were spinally hemisected at C(2), and phrenic potentials were evoked by stimulating the ventrolateral funiculus contralateral and rostral to hemisection. Phrenic potentials contralateral to the stimulating electrode were evoked at lower stimulus currents (CDR=640 +/- 46 microA; control=900 +/- 50 microA; P<0.05) and potential amplitude was significantly greater in CDR versus control rats (P<0.05). The serotonin receptor antagonist methysergide (4 mg/kg, i.v.) had no effect on the crossed phrenic potential amplitude (91+/-17% of control at 800 microA; P>0.05). Thus, CDR enhances crossed phrenic pathways but serotonin receptor activation is not necessary to maintain this effect.

Respiratory plasticity after perinatal hypercapnia in rats

Environmental conditions during early life may have profound effects on respiratory control development. We hypothesized that perinatal hypercapnia would exert lasting effects on the mammalian hypercapnic ventilatory response, but that these effects would differ between males and females. Rats were exposed to 5% CO2 from 1 to 3 days before birth through postnatal week 2 and ventilation was subsequently measured by whole-body plethysmography. In both male and female rats exposed to perinatal hypercapnia, a rapid, shallow breathing pattern was observed for the first 2 weeks after return to normocapnia, but ventilation was unchanged. Acute hypercapnic ventilatory responses (3% and 5% CO2) were reduced 27% immediately following perinatal hypercapnia, but these responses were normal after 2 weeks of recovery in both sexes and remained normal as adults. Collectively, these data suggest that perinatal hypercapnia elicits only transient respiratory plasticity in both male and female rats. This plasticity appears similar to that observed after chronic hypercapnia in adult animals and, therefore, is not unique to development.

Simultaneous MRI of lung structure and perfusion in a single breathhold

To develop and demonstrate a breathheld 3D radial ultrashort echo time (UTE) acquisition to visualize co‐registered lung perfusion and vascular structure.

64‐MULTIDETECTOR COMPUTED TOMOGRAPHIC ANGIOGRAPHY OF THE CANINE CORONARY ARTERIES

Canine coronary artery angiography (CTA) was performed in four anesthetized healthy dogs using 64-multidetector computed tomography. Esmolol, a β-1 adrenergic receptor antagonist, and sodium nitroprusside, an arteriolar and venous dilator, were administered to enhance visualization of the coronary arteries by reducing heart rate and creating vasodilation. The left main coronary artery with its three main branches and the right coronary artery were visualized and subdivided in 13 segments for evaluation. Optimal reconstruction interval, expressed as percentage of the R-to-R interval, was determined at 5% in 2.9%, 35% in 1%, 75% in 21.2%, 85% in 43.3%, and 95% in 31.7% of the segments. Overall image quality was good in 41.3% of the segments and excellent in 14.4%. There was blur in 98.1%, motion in 17.3%, and stair step in 6.7% of the evaluated segments, but these artifacts did not interfere with anatomic depiction of the arteries. Cross-sectional anatomy of the coronary arteries as evaluated from the coronary CTA agreed well with gross anatomic evaluation and published information. The use of esmolol did not lead to the target heart rate of 60-65 beats/min. Nitroprusside had no significant effect on visualized length or diameter of the coronary artery branches. Coronary CTA is useful for the anatomic depiction of coronary artery branches in the dog.

The effects of lactated Ringer’s solution (LRS) or LRS and 6% hetastarch on the colloid osmotic pressure, total protein and osmolality in healthy horses under general anesthesia

OBJECTIVE To investigate changes in colloid osmotic pressure (COP), total protein (TP) and osmolality (OSM) during anesthesia in horses given intravenous lactated Ringers solution (LRS) or LRS and hetastarch (HES). STUDY DESIGN Prospective, clinical trial. ANIMALS Fourteen horses presented for surgery. Mean age 8.3 ± 1.9 years; mean weight 452 ± 25 kg. METHODS Horses were premedicated with xylazine intravenously (IV); anesthesia was induced with ketamine and diazepam IV, and maintained with sevoflurane. Butorphanol was administered IV with pre-medications or immediately after induction. Xylazine was administered IV for recovery if necessary. LRS was administered IV to all horses with a target rate of 5-10 mL kg(-1) hour(-1). Half of the horses also received 6% HES, 2.5 mL kg(-1) over 1 hour in addition to LRS. Horses that received LRS only were considered the LRS group. Horses that received both LRS and HES were considered the LRS/HES group. Blood was drawn pre- and post-anesthesia, immediately following induction, and every 30 minutes throughout anesthesia. COP, TP and OSM were measured. RESULTS COP and TP significantly decreased at similar rates for both treatment groups from pre-anesthetic values. Pre-anesthetic COP was significantly greater in the LRS group when compared to the LRS/HES group pre-, post- and throughout anesthesia. In the LRS group post-anesthetic OSM was significantly different than the pre-anesthesia value and that for the LRS/HES group. CONCLUSIONS AND CLINICAL RELEVANCE Administration of IV HES (2.5 mL kg(-1), over 1 hour) in combination with LRS does not attenuate the decrease in COP typically seen during anesthesia with crystalloid administration alone. Based on these results, administration of HES at this rate and total volume would not be expected to prevent fluid shifts into the interstitium through its effects on COP.

p-Chlorophenylalanine eliminates long-term modulation of the exercise ventilatory response in goats.

Repeated hypercapnic exercise augments future exercise ventilatory responses, an effect termed long-term modulation. We hypothesized that serotonin depletion with p-chlorophenylalanine (PCPA, 100mg kg(-1) i.v.) would attenuate long-term modulation. Ventilation, CO(2) production and arterial blood gases were measured at rest and during exercise (4kmh(-1), 5% grade) in goats before and after training (14 hypercapnic exercise trials). Six post-training exercise trials were performed. Trials 1-3 and 4-6 were grouped for analysis (post-training 1 and 2, respectively). Without PCPA, training exaggerated the Pa(CO(2)) decrease from rest to exercise (pre-training: 1.4+/-3mmHg; post-training 1: 3.1+/-3mmHg; post-training 2: 2.3+/-3mmHg; P<0.05), indicative of long-term modulation. The Pa(CO(2)) decrease from rest to exercise was unaffected by training following PCPA (pre-training: 1.4+/-1mmHg; post-training 1: 1.4+/-3mmHg; post-training 2: 1.1+/-5mmHg; P>0.05). Thus, PCPA abolishes long-term modulation, implicating serotonin in its underlying mechanism.