Form and Function of the Vertebrate and Invertebrate Blood-Brain Barriers.

The need to protect neural tissue from toxins or other substances is as old as neural tissue itself. Early recognition of this need has led to more than a century of investigation of the blood-brain barrier (BBB). Many aspects of this important neuroprotective barrier have now been well established, including its cellular architecture and barrier and transport functions. Unsurprisingly, most research has had a human orientation, using mammalian and other animal models to develop translational research findings. However, cell layers forming a barrier between vascular spaces and neural tissues are found broadly throughout the invertebrates as well as in all vertebrates. Unfortunately, previous scenarios for the evolution of the BBB typically adopt a classic, now discredited 'scala naturae' approach, which inaccurately describes a putative evolutionary progression of the mammalian BBB from simple invertebrates to mammals. In fact, BBB-like structures have evolved independently numerous times, complicating simplistic views of the evolution of the BBB as a linear process. Here, we review BBBs in their various forms in both invertebrates and vertebrates, with an emphasis on the function, evolution, and conditional relevance of popular animal models such as the fruit fly and the zebrafish to mammalian BBB research.

Early life stress induces dysregulation of the heme pathway in adult mice.

Early life stress (ELS) is associated with cardiovascular disease (CVD) risk in adulthood, but the underlying vascular mechanisms are poorly understood. Increased hemoglobin and heme have recently been implicated to mediate endothelial dysfunction in several vascular diseases. Chronic physiological stress is associated with alterations in the heme pathway that have been well-described in the literature. However, very little is known about the heme pathway with exposure to ELS or chronic psychosocial stress. Utilizing a mouse model of ELS, maternal separation with early weaning (MSEW), we previously reported that MSEW induces endothelial dysfunction via increased superoxide production. We reasoned that heme dysregulation may be one of the culprits induced by MSEW and sustained throughout adulthood; thus, we hypothesized that MSEW induces heme dysfunction. We investigated whether circulating levels of heme, a circulating pro-oxidant mediator, are increased by MSEW and examined the role of the heme metabolic pathway and heme homeostasis in this process. We found that circulating levels of heme are increased in mice exposed to MSEW and that plasma from MSEW mice stimulated higher superoxide production in cultured mouse aortic endothelial cells (MAECs) compared to plasma from normally reared mice. The heme scavenger hemopexin blunted this enhanced superoxide production. Splenic haptoglobin abundance was significantly lower and hemoglobin levels per red blood cell were significantly higher in MSEW versus control mice. These findings lead us to propose that ELS induces increased circulating heme through dysregulation of the haptoglobin-hemoglobin system representing a mechanistic link between ELS and CVD risk in adulthood.

3D printing of face shields to meet the immediate need for PPE in an anesthesiology department during the COVID-19 pandemic.

BACKGROUND: Anesthesia providers are at risk for contracting COVID-19 due to close patient contact, especially during shortages of personal protective equipment. We present an easy to follow and detailed protocol for producing 3D printed face shields and an effective decontamination protocol, allowing their reuse. METHODS: The University of Nebraska Medical Center (UNMC) produced face shields using a combination of 3D printing and assembly with commonly available products, and produced a simple decontamination protocol to allow their reuse. To evaluate the effectiveness of the decontamination protocol, we inoculated bacterial suspensions of E. coli and S. aureus on to the face shield components, performed the decontamination procedure, and finally swabbed and enumerated organisms onto plates that were incubated for 12-24 hours. Decontamination effectiveness was evaluated using the average log10 reduction in colony counts. RESULTS: Approximately 112 face shields were constructed and made available for use in 72 hours. These methods were successfully implemented for in-house production at UNMC and at Tripler Army Medical Center (Honolulu, Hawaii). Overall, the decontamination protocol was highly effective against both E. coli and S. aureus, achieving a ≥4 log10 (99.99%) reduction in colony counts for every replicate from each component of the face shield unit. DISCUSSION: Face shields not only act as a barrier against the soiling of N95 face masks, they also serve as more effective eye protection from respiratory droplets over standard eye shields. Implementation of decontamination protocols successfully allowed face shield and N95 mask reuse, offering a higher level of protection for anesthesiology providers at the onset of the COVID-19 pandemic. CONCLUSIONS: In a time of urgent need, our protocol enabled the rapid production of face shields by individuals with little to no 3D printing experience, and provided a simple and effective decontamination protocol allowing reuse of the face shields.

Blood-brain barrier function, cell viability, and gene expression of tight junction-associated proteins in the mouse are disrupted by crude oil, benzo[a]pyrene, and the dispersant COREXIT.

Exposure to crude oil, its components, and oil dispersants during a major crude oil spill, such as the Deepwater Horizon Oil Spill, can elicit behavioral changes in animals and humans. However, the underlying mechanisms by which oil spill-related compounds alters behavior remains largely unknown. A major cause of behavioral changes generally is dysfunction of the blood-brain barrier (BBB). We investigated the impact of a crude oil high energy water accommodated fraction (HEWAF), benzo[a] pyrene (BaP; a major component of crude oil), and the oil dispersant COREXIT, on BBB function. BBB function was assessed by measuring transendothelial electrical resistance (TEER) of mouse brain microvascular endothelial cells (BMECs). Within 3 h after treatment, TEER was significantly reduced by exposure to high concentrations of all test compounds. TEER remained reduced in response to COREXIT after 48 h, but this effect waned in BMECs treated with HEWAF and BaP, with low-mid range concentrations inducing increased TEER compared to vehicle controls. At 48 h of treatment, BMEC viability was significantly reduced in response to 2% HEWAF, but was increased in response to BaP (25 and 50 µM). BMEC viability was increased with 80 ppm COREXIT, but was reduced with 160 ppm. Gene expression of tight junction-associated proteins (claudin-5 and tight junction protein-1), and cell adhesion receptor (vascular cell adhesion molecule-1) was reduced in response to HEWAF and COREXIT, but not BaP. Taken together, these data suggest that oil spill-related compounds markedly affect BBB function, and that these changes may underlie the observed behavioral changes due to crude oil exposure.

Early life stress induces immune priming in kidneys of adult male rats.

Early life stress (ELS) in humans is associated with elevated proinflammatory markers. We hypothesized that ELS induces activation of the immune response in a rat model of ELS, maternal separation (MatSep), in adulthood. MatSep involves separating pups from the dam from postnatal day 2 to postnatal day 14 for 3 h/day. Control rats are nonseparated littermates. We determined circulating and renal immune cell numbers, renal immune cell activation markers, renal cytokine levels, and the renal inflammatory gene expression response to low-dose lipopolysaccharide (LPS) in male MatSep and control rats. We observed that MatSep did not change the percentage of gated events for circulating CD3+, CD4+, CD8+, and CD4+/Foxp3+ cells or absolute numbers of mononuclear and T cells in the circulation and kidneys; however, MatSep led to an increase in activation of renal neutrophils as well as CD44+ cells. Renal toll-like receptor 4 (TLR4) and interleukin 1 beta (IL-1ß) was significantly increased in MatSep rats, specifically in the outer and inner medulla and distal nephron, respectively. Evaluation of renal inflammatory genes showed that in response to a low-dose LPS challenge (2 mg/kg iv) a total of 20 genes were significantly altered in kidneys from MatSep rats (17 genes were upregulated and 3 were downregulated), as opposed to no significant differences in gene expression in control vs. control + LPS groups. Taken together, these findings indicate that MatSep induces priming of the immune response in the kidney.

Metabolic rate and hypoxia tolerance are affected by group interactions and sex in the fruit fly (Drosophila melanogaster): new data and a literature survey.

Population density and associated behavioral adjustments are potentially important in regulating physiological performance in many animals. In r-selected species like the fruit fly (Drosophila), where population density rapidly shifts in unpredictable and unstable environments, density-dependent physiological adjustments may aid survival of individuals living in a social environment. Yet, how population density (and associated social behaviors) affects physiological functions like metabolism is poorly understood in insects. Additionally, insects often show marked sexual dimorphism (larger females). Thus, in this study on D. melanogaster, we characterized the effects of fly density and sex on both mass-specific routine oxygen consumption (V̇O2) and hypoxia tolerance (PCrit). Females had significantly lower routine V̇O2 (∼4 µl O2 mg-1 h-1) than males (∼6 µl O2 mg-1 h-1) at an average fly density of 28 flies·respirometer chamber-1 However, V̇O2 was inversely related to fly density in males, with V̇O2 ranging from 4 to 11 µl O2 mg-1 h-1 at a density of 10 and 40 flies·chamber-1, respectively (r2=0.58, P<0.001). Female flies showed a similar but less pronounced effect, with a V̇O2 of 4 and 7 µl O2 mg-1 h-1 at a density of 10 and 40 flies·chamber-1, respectively (r2=0.43, P<0.001). PCrit (∼5.5 to 7.5 kPa) varied significantly with density in male (r2=0.50, P<0.01) but not female (r2=0.02, P>0.5) flies, with higher fly densities having a lower PCrit An extensive survey of the literature on metabolism in fruit flies indicates that not all studies control for, or even report on, fly density and gender, both of which may affect metabolic measurements.

Early life stress in male mice induces superoxide production and endothelial dysfunction in adulthood.

Early life stress (ELS) is a risk for cardiovascular disease in adulthood although very little mechanistic insight is available. Because oxidative stress and endothelial dysfunction are major contributors to cardiovascular risk, we hypothesized that ELS induces endothelial dysfunction in adult male mice via increased superoxide production. Studies employed a mouse model of ELS, maternal separation with early weaning (MSEW), in which litters were separated from the dam for 4 h/day [postnatal days (PD) 2-5] and 8 h/day (PD6-16), and weaned at PD17. Control litters remained undisturbed until weaning at PD21. When compared with control mice, thoracic aortic rings from adult male MSEW mice displayed significant endothelial dysfunction that was reversed by the superoxide scavenger, polyethylene glycol-superoxide dismutase (PEG-SOD). PEG-SOD-inhibitable superoxide production by aortae from MSEW mice was significantly greater than observed in control aortae, although unaffected by nitric oxide synthase inhibition, suggesting that uncoupled nitric oxide synthase was not responsible for the accelerated superoxide production. Aortic SOD expression, plasma SOD activity, and total antioxidant activity were similar in MSEW and control mice, indicating unaltered antioxidant capacity in MSEW mice. Increased expression of the NADPH oxidase subunits, NOX2 and NOX4, was evident in the aortae of MSEW mice. Moreover, endothelial dysfunction and superoxide production in MSEW mice was reversed with the NADPH oxidase inhibitor, apocynin, indicating increased NADPH oxidase-dependent superoxide production and endothelial dysfunction. The finding that MSEW induces superoxide production and endothelial dysfunction in adult mice may provide a mechanistic link between ELS and adult cardiovascular disease risk.

Dahl SS rats demonstrate enhanced aortic perivascular adipose tissue-mediated buffering of vasoconstriction through activation of NOS in the endothelium.

Perivascular adipose tissue (PVAT) mediates buffering of vasoconstriction through activation of endothelium-derived factors. We hypothesized that the PVAT of Dahl salt-sensitive (Dahl SS) rats has reduced ability to buffer vasoconstriction. Vascular reactivity experiments were performed on aortic rings with PVAT intact (+PVAT) or removed (-PVAT), and endothelium intact (+ENDO) or removed (-ENDO) from Dahl SS rats and control SS.13(BN) rats (Dahl SS rats that have had chromosome 13 completely replaced with that of the Brown Norway rat, rendering this strain insensitive to high-salt or high-fat diet-induced hypertension). Endothelial dysfunction, assessed by ACh-mediated vasorelaxation, was confirmed in aortic rings of Dahl SS rats. The +PVAT+ENDO aortic rings had indistinguishable phenylephrine-induced vasoconstriction between genotypes. In both strains, removal of PVAT significantly enhanced vasoconstriction. Dahl SS rat -PVAT+ENDO aortic rings displayed exaggerated vasoconstriction to phenylephrine vs. SS.13(BN) rats, indicating that PVAT-mediated buffering of vasoconstriction was greater in Dahl SS rats. Removal of both the ENDO and PVAT restored vasoconstriction in both strains. The nitric oxide synthase (NOS) inhibitor, N(ω)-nitro-L-arginine methyl ester (L-NAME), produced a similar effect as that seen with -ENDO. These data indicate that the function of the PVAT to activate endothelium-derived NOS is enhanced in Dahl SS compared with SS.13(BN) rats and, most likely, occurs through a pathway that is distinct from ACh-mediated activation of NOS. PVAT weight and total PVAT leptin levels were greater in Dahl SS rats. Leptin induced a significantly decreased vasoconstriction in -PVAT+ENDO aortic rings from Dahl SS rats, but not SS.13(BN) rats. In contrast to our initial hypothesis, PVAT in Dahl SS rats buffers vasoconstriction by activating endothelial NOS via mechanisms that may include the involvement of leptin. Thus, the PVAT serves a vasoprotective role in Dahl SS rats on normal-salt diet.

Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3.

The lysine acetylation state of nonhistone proteins may be regulated through histone deacetylases (HDACs). Evidence suggests that nitric oxide (NO) synthase 3 (NOS3; endothelial NOS) is posttranslationally lysine acetylated, leading to increased NO production in the endothelium. We tested the hypothesis that NOS3 is lysine acetylated and that upregulated HDAC1-mediated deacetylation leads to reduced NO production in endothelial cells. We determined that NOS3 is basally lysine acetylated in cultured bovine aortic endothelial cells (BAECs). In BAECs, HDAC1 is expressed in the nucleus and cytosol and forms a novel protein-protein interaction with NOS3. Overexpression of HDAC1 in BAECs resulted in a significant reduction in NOS3 lysine acetylation (control = 1.0 ± 0.1 and HDAC1 = 0.59 ± 0.08 arbitrary units, P < 0.01) and significantly blunted basal nitrite production (control 287.7 ± 29.1 and HDAC1 172.4 ± 31.7 pmol·mg(-1)·h(-1), P < 0.05) as well as attenuating endothelin-1-stimulated nitrite production (control = 481.8 ± 50.3 and HDAC1 243.1 ± 48.2 pmol·mg(-1)·h(-1), P < 0.05). While HDAC1 knockdown with small-interfering RNA resulted in no change in NOS3 acetylation level, yet increased basal nitrite production (730.6 ± 99.1 pmol·mg(-1)·h(-1)) and further exaggerated increases in endothelin-1 stimulated nitrite production (1276.9 ± 288.2 pmol·mg(-1)·h(-1)) was observed. Moreover, overexpression or knockdown of HDAC1 resulted in no significant effect on NOS3 protein expression or NOS3 phosphorylation sites T497, S635, or S1179. Thus these data indicate that upregulated HDAC1 decreases NOS3 activity, most likely through direct lysine deacetylation of NOS3. We propose that HDAC1-mediated deacetylation of NOS3 may represent a novel target for endothelial dysfunction.

Transgenerational epigenetics: the role of maternal effects in cardiovascular development.

Transgenerational epigenetics, the study of non-genetic transfer of information from one generation to the next, has gained much attention in the past few decades due to the fact that, in many instances, epigenetic processes outweigh direct genetic processes in the manifestation of aberrant phenotypes across several generations. Maternal effects, or the influences of maternal environment, phenotype, and/or genotype on offsprings' phenotypes, independently of the offsprings' genotypes, are a subcategory of transgenerational epigenetics. Due to the intimate role of the mother during early development in animals, there is much interest in investigating the means by which maternal effects can shape the individual. Maternal effects are responsible for cellular organization, determination of the body axis, initiation and maturation of organ systems, and physiological performance of a wide variety of species and biological systems. The cardiovascular system is the first to become functional and can significantly influence the development of other organ systems. Thus, it is important to elucidate the role of maternal effects in cardiovascular development, and to understand its impact on adult cardiovascular health. Topics to be addressed include: (1) how and when do maternal effects change the developmental trajectory of the cardiovascular system to permanently alter the adult's cardiovascular phenotype, (2) what molecular mechanisms have been associated with maternally induced cardiovascular phenotypes, and (3) what are the evolutionary implications of maternally mediated changes in cardiovascular phenotype?

Parental hypoxic exposure confers offspring hypoxia resistance in zebrafish (Danio rerio).

Parental influences are a potentially important component of transgenerational transfer of phenotype in vertebrates. This study examined how chronic hypoxic exposure on adult zebrafish (Danio rerio) affected the phenotype of their offspring. Separate adult populations were exposed to hypoxia (13.1 kPa O(2)) or normoxia (21.1 kPa O(2)) for periods ranging from 1 to 12 weeks. Adults were then returned to normoxia and bred within experimental groups. Adult fecundity and egg characteristics (volume of egg, yolk and perivitelline fluid) were assessed. Subsequently, larval body length, time to loss of equilibrium in severe hypoxia (~4 kPa O(2)), and critical thermal minima (CT(min)) and maxima (CT(max)) were measured at 6, 9, 12, 15, 18, 21 and 60 days post-fertilization (d.p.f.). Adult fecundity was depressed by hypoxic exposure. Egg component volumes were also depressed in adults exposed to 1-2 weeks of hypoxia, but returned to control levels following longer hypoxic exposure. Adult hypoxic exposures of >1 week resulted in longer body lengths in their larval offspring. Time to loss of equilibrium in severe hypoxia (i.e. hypoxic resistance) in control larvae decreased from 6 to 12 d.p.f., remaining constant thereafter. Notably, hypoxic resistance from 6 to 18 d.p.f. was ~15% lower in larvae whose parents were exposed to just 1 week of chronic hypoxia, but resistance was significantly increased by ~24-30% in 6-18 d.p.f. larvae from adults exposed to 2, 3 or 4 weeks of hypoxia. CT(min) (~10-12°C) and CT(max) (~39.5°C) were unchanged by parental hypoxic exposure. This study demonstrates that parental hypoxic exposure in adult zebrafish has profound epigenetic effects on the morphological and physiological phenotype of their offspring.

Changing standard chow diet promotes vascular NOS dysfunction in Dahl S rats.

We hypothesized that vascular nitric oxide synthase (NOS) function and expression is differentially regulated in adult Dahl salt-sensitive rats maintained on Teklad or American Institutes of Nutrition (AIN)-76A standard chow diets from 3 to 16 wk old. At 16 wk old, acetylcholine (ACh)-mediated vasorelaxation and phenylephrine (PE)-mediated vasoconstriction in the presence and absence of NOS inhibitor, N(ω)-nitro-L-arginine methyl ester (L-NAME), was assessed in small-resistance mesenteric arteries and aortas. Rats maintained on either diet throughout the study had similar responses to ACh and PE in the presence or absence of L-NAME in both vascular preparations. We reasoned that changing from one diet to another as adults may induce vascular NOS dysfunction. In the absence of L-NAME, small arteries from Teklad-fed rats switched to AIN-76 diet and vice versa had similar responses to ACh and PE. Small-arterial NOS function was maintained in rats switched to AIN-76A from Teklad diet, whereas NOS function in response to ACh and PE was lost in the small arteries from rats changed to Teklad from AIN-76A diet. This loss of NOS function was echoed by reduced expression of NOS3, as well as phosphorylated NOS3. The change in NOS phenotype in the small arteries was observed without changes in blood pressure. Aortic responses to ACh or PE in the presence or absence of L-NAME were similar in all diet groups. These data indicate that changing standard chow diets leads to small arterial NOS dysfunction and reduced NOS signaling, predisposing Dahl salt-sensitive rats to vascular disease.

Egg yolk environment differentially influences physiological and morphological development of broiler and layer chicken embryos.

Maternal effects are important in epigenetic determination of offspring phenotypes during all life stages. In the chicken (Gallus gallus domesticus), transgenerational transfer of egg yolk factors may set the stage for morphological and physiological phenotypic differences observed among breeds. To investigate the effect of breed-specific yolk composition on embryonic broiler and layer chicken phenotypes, we employed an ex ovo, xenobiotic technique that allowed the transfer of broiler and layer chicken embryos from their natural yolks to novel yolk environments. Embryonic day two broiler embryos developing on broiler yolk culture medium (YCM) had significantly higher heart rates than layer embryos developing on layer YCM (176±7 beats min(-1) and 147±7 beats min(-1), respectively). Broiler embryos developing on layer YCM exhibited heart rates typical of layer embryos developing normally on layer YCM. However, layer embryo heart rates were not affected by development on broiler YCM. Unlike O(2) consumption, development rate and body mass of embryos were significantly affected by exposure to different yolk types, with both broiler and layer embryos displaying traits that reflected yolk source rather than embryo genotype. Analysis of hormone concentrations of broiler and layer egg yolks revealed that testosterone concentrations were higher in broiler yolk (4.63±2.02 pg mg(-1) vs 3.32±1.92 pg mg(-1)), whereas triiodothyronine concentrations were higher in layer yolk (1.05±0.18 pg mg(-1) vs 0.46±0.22 pg mg(-1)). Thus, a complex synergistic effect of breed-specific genotype and yolk environment exists early in chicken development, with yolk thyroid hormone and yolk testosterone as potential mediators of the physiological and morphological effects.

Differential impact of cocaine on meal patterns in female and male rats.

Female rats, relative to males, exhibit greater behavioral activation to cocaine and other psychostimulants, but the effect of sex and the estrous cycle in modulating the hypophagic action of cocaine has not been evaluated. Meal patterns were recorded in automated food hoppers during the first 3 h of the dark phase in adult female and male rats after administration of ascending cocaine doses (0, 7.5, and 15 mg/kg cocaine, i.p.) on successive trials. Cocaine produced a greater suppression of feeding as well as a reduction in meal number over a 3 h test period in female rats during estrus, relative to that noted during diestrus. In contrast, during the 180 min test period, male rats showed minimal hypophagic responses to 7.5 or 15 mg/kg cocaine. These results extend the range of behavioral perturbations induced by cocaine that are modulated by sex and by the estrous cycle and are consistent with the notion that estradiol may modulate the neurochemical actions of cocaine.

Meal patterns and body weight after nicotine in male rats as a function of chow or high-fat diet.

Studies of the effects of nicotine (NIC) on meal patterns in rats often employ chow pellet diets that contain little fat, whereas humans using NIC commonly consume diets relatively rich in fat. The aim of the present study was therefore to compare the impact of NIC administration and NIC cessation on meal pattern in adult male rats offered a standard powdered chow (CHOW: 10.9% fat by calories) diet or a palatable high-fat (HIFAT: 58.3% fat by calories) diet. Computerized meal pattern analyses were conducted for male rats treated for 14 days with injections of either saline or 1.4 mg/kg/day of NIC (as the free base given in 5 equal amounts) during the dark phase and continued for 10 days after NIC cessation. The suppression of daily caloric intake by NIC was larger in HIFAT-NIC rats than in CHOW-NIC rats (p < .01), such that NIC induced a greater suppression of body weight in HIFAT-NIC rats, relative to CHOW-NIC rats (p < 0.02). NIC administration reduced MS in both CHOW and HIFAT rats. CHOW fed rats showed a gradual increase in meal number in response to NIC, whereas HIFAT fed rats showed a significant initial suppression of meal number, which returned to control levels by day 4 of the 14 day NIC treatment period. In addition, NIC increased water intake more in HIFAT fed rats than in CHOW rats. Cessation of NIC resulted in transient increases in daily caloric intake in CHOW and in HIFAT rats. The present study demonstrates that NIC actions on food intake suppression, meal patterns, and weight reduction differ depending on whether the rats are fed low- or high-fat diets.

Concurrent measures of feeding and locomotion in rats.

Psychostimulants including amphetamine and cocaine induce locomotion and stereotypy and suppress eating. Studies of the biobehavioral actions of psychostimulants commonly focus on locomotion and less commonly on feeding, and only rarely are these measures considered in conjunction within the same animal. Inasmuch as hyperactivity induced by a psychostimulant may compete with other motor behaviors, including eating, it would be important to concurrently assess changes in eating and locomotion after psychostimulant treatment. The present paper describes a modification of an automated activity chamber in which minute-by-minute recordings of food consumption are gathered in parallel with an assessment of locomotion. The present experiment illustrates the method by characterizing the temporal changes in locomotion and eating produced by administration of hypophagic doses of nicotine tartrate (0.28 mg/kg (as the base), IP) or cocaine hydrochloride (7.5 mg/kg, IP). At these doses, nicotine suppressed eating and locomotion, whereas cocaine suppressed eating, but facilitated forward locomotion. These outcomes support the viability of this apparatus and the concurrent method for the dissociation of feeding and locomotion.

An inexpensive food cup for use in a commercially available food monitoring system.

The microstructure of feeding in rats can be probed using a variety of protocols that employ videotape-based ratings, pellet feeders, and/or laboratory balances. A recent commercial product (BioDAQ, Research Diets, New Brunswick, NJ) uses a metal food hopper placed on a load cell to monitor daily food pellet consumption. In this system, movements of the hopper during eating and the cessation of hopper movements after eating are combined with momentary hopper weights for subsequent analyses of daily meal patterns. Our laboratory has devised an improved food cup for the BioDAQ system that is easily balanced, minimizes spillage, and is compatible with either powdered chow diets or nonpelleted soft diets (e.g., a high-fat diet). In the present paper, we describe the methods used to fabricate this food cup and present data illustrating its use in meal pattern analyses for rats fed either a ground laboratory chow or a 33% high-fat diet.

Noradrenergic modulation of ephedrine-induced hypophagia.

The hypophagic action of the sympathomimetic amine ephedrine (EPH) in the rat may reflect actions on central dopaminergic (DA) and noradrenergic (NE) systems. EPH indirectly facilitates DA and NE activity and acts as a partial agonist at alpha(1)-adrenergic receptors. Two approaches were used to assess the possible contribution of NE and DA pathways to EPH-induced hypophagia. In the first, regression analyses of published archival data were computed to characterize the relation between the hypophagic potency values of (-)-(EPH) and related sympathomimetic drugs, including (+)-amphetamine, aminorex, mazindol, and phentermine (data derived from Blosser JC et al., 1987) and the most potent action of these drugs on facilitating NE activity or DA activity in rat brain (data derived from Rothman RB et al., 2001). In the NE analyses, the ED(50) values for these drugs for the inhibition of eating in rats were significantly related (r = 0.91, P = 0.03) to the potency of each drug in facilitating NE activity (either release or inhibition of [(3)H]NE reuptake), whereas in the DA analyses the correlation between ED(50) values and DA activity for these drugs was also significant (r = 0.98, P = 0.003). The regression analyses are thus supportive of a role for NE or DA in the hypophagic capacity of EPH. Although an earlier study noted that administration of the putative DA antagonist pimozide in rats attenuated EPH hypophagia, pimozide exerts similar potency in antagonizing DA receptors and alpha(1)-adrenergic receptors. To clarify the role of alpha(1)-adrenoceptors in EPH-induced hypophagia, adult male rats were pretreated with the alpha(1)-adrenergic receptor antagonist prazosin (0.0.5 and 2 mg/kg) prior to the administration of (-)-EPH (0, 5, 10, or 20 mg/kg, IP). Prazosin pretreatment at 2.0 mg/kg significantly attenuated the hypophagia, but not the hypodipsia, induced by administration of 10 mg/kg and by 20 mg/kg (-)-EPH. Collectively, these results confirm a critical contribution of of alpha(1)-adrenoceptors to the hypophagic action of (-)-EPH in rats.