Juvenile Spontaneously Hypertensive Rats Research Articles

Subdiaphragmatic vagal nerve stimulation attenuates the development of hypertension and alters nucleus of the solitary tract transcriptional networks in the spontaneously hypertensive rat.

Augmented vagal signaling may be therapeutic in hypertension. Most studies to date have used stimulation of the cervical vagal branches. Here, we investigated the effects of chronic intermittent electric stimulation of the ventral subdiaphragmatic vagal nerve branch (sdVNS) on long-term blood pressure, immune markers, and gut microbiota in the spontaneously hypertensive rat (SHR), a rodent model of hypertension characterized by vagal dysfunction, gut dysbiosis, and low-grade inflammation. We evaluated the effects of sdVNS on transcriptional networks in the nucleus of the solitary tract (NTS), a major cardioregulatory brain region with direct gut vagal projections. Male juvenile SHRs were implanted with radiotelemetry transmitters and vagal nerve cuffs for chronic intermittent electric sdVNS, applied three times per day for 7 consecutive weeks followed by 1 wk of no stimulation. Blood pressure was measured once a week using telemetry in the sdVNS group as well as age-matched sham-stimulated SHR controls. At the endpoint, colonic and circulating inflammatory markers, corticosterone, and circulating catecholamines were investigated. Bacterial 16 s sequencing measured gut bacterial abundance and composition. RNA sequencing evaluated the effects of sdVNS on transcriptional networks in the NTS. SHRs that received sdVNS exhibited attenuated development of hypertension compared with sham animals. No changes in peripheral inflammatory markers, corticosterone, or catecholamines and no major differences in gut bacterial diversity and composition were observed following sdVNS, apart from decreased abundance of Defluviitaleaceale bacterium detected in sdVNS SHRs compared with sham animals. RNA sequencing revealed significant sdVNS-dependent modulation of select NTS transcriptional networks, including catecholaminergic and corticosteroid networks.NEW & NOTEWORTHY We show that stimulation of the ventral subdiaphragmatic vagal nerve branch may be a promising potential approach to treating hypertension. The data are especially encouraging given that rodents received only 30 min per day of intermittent stimulation therapy and in view of the potential of long-term blood pressure effects that are not stimulus-locked.

Effects of Neuroinflammation and Autophagy on the Structure of the Blood-Brain Barrier in ADHD Model.

Spontaneously hypertensive rats (SHR) are the most common animal model used to study attention deficit hyperactivity disorder (ADHD). The purpose of this study was to look at the impact of neuroinflammation and autophagy on blood-brain barrier function in the prefrontal cortex and hippocampus of ADHD rats. The rats were separated into three groups: juvenile SHR (6weeks), mature SHR (12weeks), and comparable age WKY groups. An open-field test was used to assess rats' ability to move on their own. Immunofluorescence was used to detect the Iba1-immunopositive microglia, ZO-1 and TNF-α. Meanwhile, the expression of p62, Beclin-1, LC3B, and MMP9, MMP2, TNF-α, ZO-1, and occludin were detected by Western blot. The results have shown that Iba1-immunopositive microglia and TNF-α protein in the brain of SHR rats were significantly increased. Moreover, autophagy of cells and the level of MMP2 and MPP9 in the prefrontal cortex and hippocampus increased in SHR rats. In addition, the expression of ZO-1 and occludin was decreased in SHR rats. To sum up, the increase of neuroinflammation and excessive autophagy were essential factors for the damage of blood-brain barrier structure and function.

The mesocortical dopaminergic system cannot explain hyperactivity in an animal model of attention deficit hyperactivity disorder (ADHD)- Spontaneously hypertensive rats (SHR)

BackgroundAttention deficit hyperactivity disorder (ADHD) is one of the most prevalent neuropsychiatric disorders with morphological brain abnormalities. There is a growing body of evidence that abnormalities in the dopaminergic system may account for ADHD pathogenesis. However, it is not clear whether the dopaminergic system is hyper or hypoactive. To determine whether the DA neurons and/or axons deficiency might be the cause of the postulated dopaminergic hypofunction in spontaneously hypertensive rats (SHR, animal model of ADHD), this study examined the dopaminergic neurons and fibers in the brain tissues of SHRs and Wistar Kyoto rats (WKY, control animals). Here, we performed immunohistochemical tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH) staining on brain sections collected on juveniles from SHR and WKY. Moreover, behavioral testing to examine the hyperactivity in the open field area was also elucidated.ResultsThe mesocortical dopaminergic system appears to be normal in juvenile SHR, as suggested by (i) no alteration in the area density of TH-immunoreactive (TH-ir) dopaminergic neurons in the ventral tegmental area (VTA), (ii) no alterations in the volume density of TH-ir fibers in layer I of the prelimbic (PrL) subregion of medial PFC (mPFC), (iii) no alteration in the percentage of TH-ir dopaminergic fibers in layer I of the PrL subregion of mPFC as revealed by TH and/or DBH immunoreactivity. Furthermore, the SHR showed increased locomotor activity than WKY in the open field test.ConclusionsThe demonstration of no alteration in mesocortical dopaminergic neurons and fiber in SHR raises some concern about the position of SHR as an animal model of the inattentive subtype of ADHD. However, these results strengthen this strain as an animal model of hyperactive/impulsive subtype ADHD for future studies that may elucidate the underlying mechanism mediating hyperactivity and test various treatment strategies.

Effects of Long-Term Intervention with Losartan, Aspirin and Atorvastatin on Vascular Remodeling in Juvenile Spontaneously Hypertensive Rats

Hypertension in adolescents is associated with adverse cardiac and vascular events. In addition to lowering blood pressure, it is not clear whether pharmacological therapy in early life can improve vascular remodeling. This study aimed to evaluate the effects of long-term administration of losartan, aspirin, and atorvastatin on vascular remodeling in juvenile spontaneously hypertensive rats (SHRs). Losartan, aspirin, and atorvastatin were administered via gavage at doses of 20, 10, and 10 mg/kg/day, respectively, on SHRs aged 6-22 weeks. Paraffin sections of the blood vessels were stained with hematoxylin-eosin (H&E) and Sirius Red to evaluate the changes in the vascular structure and the accumulation of different types of collagen. The plasma levels of renin, angiotensin II (Ang II), aldosterone (ALD), endothelin-1 (ET-1), interleukin-6 (IL-6), and neutrophil elastase (NE) were determined using ELISA kits. After the 16-week treatment with losartan, aspirin, and atorvastatin, the wall thickness of the thoracic aorta and carotid artery decreased. The integrity of the elastic fibers in the tunica media was maintained in an orderly manner, and collagen deposition in the adventitia was retarded. The plasma levels of renin, ALD, ET-1, IL-6, and NE in the SHRs also decreased. These findings suggest that losartan, aspirin, and atorvastatin could improve vascular remodeling beyond their antihypertensive, anti-inflammatory, and lipid-lowering effects. Many aspects of the protection provided by pharmacological therapy are important for the prevention of cardiovascular diseases in adults and older adults.

Paradoxical Increase in Body Mass Induced by Beta-Guanidinopropionic Acid in Juvenile Spontaneously Hypertensive Rats.

BackgroundThe adenosine triphosphate (ATP) regenerating enzyme creatine kinase (CK) is intimately involved in blood pressure generation. Consequently, the creatine transporter and CK inhibitor beta-guanidinopropionic acid (GPA) successfully reduced blood pressure in 16-week-old spontaneously hypertensive rats (SHR), but GPA may cause growth retardation in juvenile mammals. This report considers a serendipity observation of paradoxical growth increase after using GPA to prevent hypertension in three-week-old SHR.MethodsImplementing the “Animal Research: Reporting of In Vivo Experiments” (ARRIVE) guideline, male, three-week-old spontaneously hypertensive rats (N=22) were randomly assigned to standard soy-based (creatine-free) chow with GPA 0.1% vs control chow during four weeks (primary, t=4w) or six weeks of treatment (t=6w). Blood pressure measured by the tail-cuff method was the main outcome. Other outcomes included body mass and contractility characteristics of isolated arteries.Results Body mass at baseline was 28.4 (SE 0.71) g (n=22). With similar food intake/100 gram animal in both groups, GPA-treated rats (n=11) developed a strikingly larger body size and mass: t=4w, GPA 110.4 g (3.7) vs controls (n=11) 65.0 g (4.8) (+69.8%; p<0.001); t=6w, GPA 154.3 (4.7) vs controls 68.0 (4.7) g. There were no significant differences in cardiovascular parameters including blood pressure.DiscussionAn unexpected increase in body mass and size without concurrent blood pressure increase was observed in juvenile SHR on GPA vs control soy-based chow. It is speculated that the partial creatine agonist activity of GPA contributed to these effects. Further studies are needed to confirm these findings and better understand the impact of modulating energy metabolism in juvenile hypertension-prone mammals.

The cytokine alterations/abnormalities and oxidative damage in the pancreas during hypertension development

The aim of the present study was to compare the content of cytokines, chemokines, and oxidative stress markers in the pancreas of spontaneously hypertensive rats (SHRs) and Wistar Kyoto Rats (WKYs) serving as controls. Enzyme-like immunosorbent assay (ELISA) and biochemical methods were used to measure pancreatic levels of interleukin-1ß, interleukin-6, tumor necrosis factor α, transforming growth factor β, RANES, monocyte chemoattractant protein 1, interferon gamma-induced protein 10, malondialdehyde, and sulfhydryl groups. The results showed that the pancreatic concentrations of all studied cytokines and chemokines did not differ between 5-week-old SHRs and WKYs, except RANTES which was significantly reduced in juvenile SHRs. In 10-week-old animals, except interleukin-1ß, the levels of all these proteins were significantly reduced in SHRs. The pancreatic levels of malondialdehyde were significantly reduced in 5-week-old SHRs and significantly elevated in 10-week-old SHRs while the contents of sulfhydryl groups were similar in both rat strains at any age studied. In conclusion, these data provide evidence that in maturating SHRs, the pancreatic levels of cytokines and chemokines are significantly reduced, while malondialdehyde significantly elevated. This suggests that in the pancreas of mature SHRs, the inflammation process is suppressed but there is ongoing oxidative damage.

Age‐dependent Changes of CO2 Chemoreflex and Blood Pressure (BP) in Juvenile and Adult Spontaneously Hypertensive Rats (SHRs)

Activation of central chemoreceptors by CO2 increases BP, sympathetic nerve activity (SNA), and breathing. Experiments using SHRs, a well‐established neurogenic hypertensive model, have shown that the high BP is associated with elevated SNA and peripheral chemoreflexes, and an impaired baroreflex. Earlier we showed that adult SHRs have enhanced ventilatory (VE) and BP responses to hypercapnia, and blocking both orexin receptors can normalize such enhanced responses and markedly lower BP. Here, using juvenile SHRs (P42‐60), we report that: 1) VE responses to normoxic‐hypercapnia and hyperoxic‐hypercapnia (179±25 &amp; 162±38 %increase respectively) are higher than that of age‐matched normal rats (118±22 &amp; 113±26 %increase, respectively), but lower than that of adult SHRs (226±39 % increase)(P&lt;0.05); 2) resting BP is higher than of age‐matched normal rats (130±12 &amp; 99±5mmHg respectively), but lower than that of adult SHRs (157±12mmHg)(P&lt;0.05). Summary, in SHRs 1) significant increases in both CO2 chemoreflex and BP are measureable in juveniles; 2) the enhanced CO2 chemoreflex persists while peripheral chemoreceptors were inhibited by hyperoxia indicate an important role for central chemoreflex in neurogenic hypertension. We suggest that the enhanced CO2 chemoreflex in early life may be causally linked to the development of neurogenic hypertension in SHRs, and modulation of the overactive CO2 chemoreflex could be beneficial in treating neurogenic hypertension. (HL 28066)

Stimulation of postsynapse adrenergic α2A receptor improves attention/cognition performance in an animal model of attention deficit hyperactivity disorder

A 5-trial inhibitory avoidance test using spontaneously hypertensive rat (SHR) pups has been used as an animal model of attention deficit hyperactivity disorder (ADHD). However, the roles of noradrenergic systems, which are involved in the pathophysiology of ADHD, have not been investigated in this model. In the present study, the effects of adrenergic α2 receptor stimulation, which has been an effective treatment for ADHD, on attention/cognition performance were investigated in this model. Moreover, neuronal mechanisms mediated through adrenergic α2 receptors were investigated. We evaluated the effects of both clonidine, a non-selective adrenergic α2 receptor agonist, and guanfacine, a selective adrenergic α2A receptor agonist, using a 5-trial inhibitory avoidance test with SHR pups. Juvenile SHR exhibited a shorter transfer latency, compared with juvenile Wistar Kyoto (WKY) rats. Both clonidine and guanfacine significantly prolonged the transfer latency of juvenile SHR. The effects of clonidine and guanfacine were significantly blocked by pretreatment with an adrenergic α2A receptor antagonist. In contrast, the effect of clonidine was not attenuated by pretreatment with an adrenergic α2B receptor antagonist, or an adrenergic α2C receptor antagonist, while it was attenuated by a non-selective adrenergic α2 receptor antagonist. Furthermore, the effects of neither clonidine nor guanfacine were blocked by pretreatment with a selective noradrenergic neurotoxin. These results suggest that the stimulation of the adrenergic α2A receptor improves the attention/cognition performance of juvenile SHR in the 5-trial inhibitory avoidance test and that postsynaptic, rather than presynaptic, adrenergic α2A receptor is involved in this effect.

Atomoxetine-Induced Increases in Monoamine Release in the Prefrontal Cortex are Similar in Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

Spontaneously hypertensive rats (SHRs) are used as a model for attention-deficit/hyperactivity disorder (ADHD), since SHRs are hyperactive and show defective sustained attention in behavioral tasks. The psychostimulants amphetamine and methylphenidate and the selective norepinephrine reuptake inhibitor atomoxetine are used as ADHD medications. The effects of high K(+) stimulation or psychostimulants on brain norepinephrine or dopamine release in SHRs have been previously studied both in vitro and in vivo, but the effects of atomoxetine on these neurotransmitters have not. The present study examined the effects of administration of atomoxetine on extracellular norepinephrine, dopamine, and serotonin levels in the prefrontal cortex of juvenile SHRs and Wistar-Kyoto (WKY) rats. Baseline levels of prefrontal norepinephrine, dopamine, and serotonin were similar in SHRs and WKY rats. Systemic administration of atomoxetine (3 mg/kg) induced similar increases in prefrontal norepinephrine and dopamine, but not serotonin, levels in both strains. Furthermore, there was no difference in high K(+)-induced increases in extracellular norepinephrine, dopamine, and serotonin levels in the prefrontal cortex between SHRs and WKY rats. These findings indicate that monoamine systems in the prefrontal cortex are similar between SHRs and WKY rats.

Persistent α1-adrenergic receptor function in the nucleus locus coeruleus causes hyperexcitability in AD/HD model rats

Spontaneously hypertensive rats (SHR) are widely used as a model of attention deficit hyperactivity disorder (ADHD) as their ADHD-like behaviors are restored by methylphenidate. However, a postnatal neural development in SHR is unknown. We performed whole cell patch clamp recordings from locus coeruleus (LC) neurons in neonatal [postnatal day (P) 3-5], juvenile (P21-28), and adult (P 49-56) SHR and age-matched Wistar rats to evaluate α1- and α2-adrenergic receptor (ARs) activities at each developmental period. LC neurons in neonatal Wistar rats and SHR showed no difference in resting membrane potential and spontaneous firing rate, while juvenile and adult SHR LC neurons showed depolarized resting membrane potential and faster spontaneous firing rate than in Wistar rats. Blockade of α1-AR activity by prazosin hyperpolarized the membrane and abolished spontaneous firings in all developmental periods in SHR LC neurons, but not in juvenile and adult Wistar rats. α1-AR stimulation by phenylephrine evoked an inward current in juvenile LC neurons in SHR, but not in juvenile Wistar rats. This phenylephrine-induced inward current was abolished by nonselective cation channel blockers. By contrast, α2-AR stimulation-induced outward currents in the presence of an α1-AR antagonist were equivalent in SHR and Wistar LC neurons. These data suggest that Wistar LC neurons lose α1-AR function during development, whereas α1-ARs remain functional in SHR LC neurons. Thus persistent intrinsic activity of α1-ARs may be a neural mechanism contributing to developmental disorders in juvenile SHRs.

Intra-orbitofrontal cortex injection of haloperidol removes the beneficial effect of methylphenidate on reversal learning of spontaneously hypertensive rats in an attentional set-shifting task

Numerous studies suggest that attention-deficit/hyperactivity disorder (ADHD) is caused by deficits in catecholaminergic systems. Furthermore, dysfunctions of prefrontal cortex can impair inhibitory controls of ADHD patients, resulting in their impulsive behaviors. Researchers also find that rats with lesions in the orbitofrontal cortex show deficits in the reversal learning of attentional set-shifting task (ASST), a behavioral test frequently used in human studies to asses the inhibition system. However, the role of orbitofrontal dopamine system in the mechanism responsible for the dysfunctions of inhibitory controls in ADHD patients and animal models remains unknown. In the present study, we manipulated orbitofrontal dopamine activities of spontaneously hypertensive rats (SHR), a widely used ADHD animal model, through intra-peritoneal injection of methylphenidate (MPH) and central infusion of haloperidol, and observed their performances in ASST. The results show that juvenile SHRs learned slower than Wistar controls in the first and second reversal learnings of ASST. The deficits could be removed by intra-peritoneal injections of MPH. Furthermore, central infusions of haloperidol in the orbitofrontal cortex blocked the effects of MPH. In conclusions, dopamine activity in orbitofrontal cortex might play a crucial role in the neural mechanism of reversal learning deficits in this animal model of ADHD.

Upregulation of junctional adhesion molecule-A is a putative prognostic marker of hypertension

AimsEstablishing biochemical markers of pre-hypertension and early hypertension could help earlier diagnostics and therapeutic intervention. We assess dynamics of junctional adhesion molecule-A (JAM-A) expression in rat models of hypertension and test whether JAM-A expression could be driven by angiotensin (ANG) II and whether JAM-A contributes to the progression of hypertension. We also compare JAM-A expression in normo- and hypertensive humans.Methods and resultsIn pre-hypertensive and spontaneously hypertensive rats (SHRs), JAM-A protein was overexpressed in the brainstem microvasculature, lung, liver, kidney, spleen, and heart. JAM-A upregulation at early and late stages was even greater in the stroke-prone SHR. However, JAM-A was not upregulated in leucocytes and platelets of SHRs. In Goldblatt 2K-1C hypertensive rats, JAM-A expression was augmented before any increase in blood pressure, and similarly JAM-A upregulation preceded hypertension caused by peripheral and central ANG II infusions. In SHRs, ANG II type 1 (AT1) receptor antagonism reduced JAM-A expression, but the vasodilator hydralazine did not. Body-wide downregulation of JAM-A with Vivo-morpholinos in juvenile SHRs delayed the progression of hypertension. In the human saphenous vein, JAM-A mRNA was elevated in hypertensive patients with untreated hypertension compared with normotensive patients but reduced in patients treated with renin–angiotensin system antagonists.ConclusionBody-wide upregulation of JAM-A in genetic and induced models of hypertension in the rat precedes the stable elevation of arterial pressure. JAM-A upregulation may be triggered by AT1 receptor-mediated signalling. An association of JAM-A with hypertension and sensitivity to blockers of ANG II signalling were also evident in humans. We suggest a prognostic and possibly a pathogenic role of JAM-A in arterial hypertension.

Region-specific elevation of D1 receptor-mediated neurotransmission in the nucleus accumbens of SHR, a rat model of attention deficit/hyperactivity disorder

Spontaneously hypertensive rats (SHR) are widely used as a rat model of attention deficit/hyperactivity disorder (AD/HD). Here, we conducted neurochemical and behavioral studies in SHR to clarify the topographical alterations in neurotransmissions linked to their behavioral abnormalities. In the open-field test, juvenile SHR showed a significant hyperactivity in ambulation and rearing as compared with Wistar Kyoto rats (WKY). Brain mapping analysis of Fos-immunoreactivity (IR) revealed that SHR showed a marked increase in Fos expression in the core part (AcC) of the nucleus accumbens (NAc). Small to moderate increases were also observed in the shell part of the NAc and some regions of the cerebral cortex (e.g., parietal association cortex). These changes in Fos expression were region-specific and the Fos-IR levels in other brain regions (e.g., hippocampus, amygdala, striatum, thalamus and hypothalamus) were unaltered. In addition, treatment of SHR with the selective D1 antagonist SCH-23390 significantly reversed both behavioral hyperactivity and elevated Fos expression in the AcC and cerebral cortex. The present study suggests that D1 receptor-mediated neurotransmission in the AcC is region-specifically elevated in SHR, which could be responsible for behavioral hyperactivity.

The effects of ritalin on the both attentional set shifting and levels of neurotransmitters and their metabolites in the mPFC in rat as a model of attention deficit hyperactivity disorder

Objective To evaluate the enhancement effects of ritalin enhancement of attentional set shifting and motor activity by assessing ability of attentional set shifting in juvenile spontaneously hypertensive rats (SHRs)as an animal model of attention deficit and hyperactivity disorder.Methods Sixteen male 4 week-old SHRs were randomly divided into titalin(SR)and saline(SN),control groups(WN)respectively.The number of trails were recorded as the sign of the ability of attentional set shifting.Results The number of trails to reach the criterion of six consecutive was significantly more in the SN group than those of WN group in SD,CD,CDR,IDS,EDS(P＜0.05,P＜0.01).The same was observed in the SN group than in the ritalin-treated group(SR)in all stages except IDSR(P＜0.05,P＜0.01).Ventral subdivisions of mPFC:(1)At the base line,SN((504.142±100.96)ng·g~(-1))group have exhibited a higher NE((354.42±125.38)ng·g~(-1),P=0.001)level,MHPG/DOPAC ratio,a lower utilization ratio(MHPG/NE=3.28±1.11,DOPAC/DA=0.93±0.31)and a lower MHPG level((1560.04±159.69)ng·g~(-1))when compared with the WN subgroup((1709.95±322.29)ng·g~(-1),P=0.000).(2)The ritalin treatment in SHRs decreased MHPG/DOPAC ratio and also increased the utilization ratio(MHPG/NE:1.95±0.37,3.28±1.11,P=0.000;DOPAC/DA:0.31±0.15,0.93±0.31,P=0.000),and MHPG,DOPAC level when compared with the SN group(P＜0.05～P＜0.01).Conclusions(1)The juvenile SHRs have the disability of attentional set shifting,ritalin has shown effect to improve the ability of attentional set shifting in SHRs.(2)Decreased utilization ratios(DOPAC/DA,MHPG/NE)were found in in the ventral subdivision of mPFC in the SHRs while a relative unbalance between hypodopaminergic and hypemoradrenergic activity was also noted.(3)Effects of ritalin has been observed in enhancement of cognitive flexibility by way of increasing the utilization ratios(DOPAC/DA,MHPG/NE)and hence the relative unbalance hypodopaminergic and hypemoradrenergic activity is improved. Key words: Attentional set shifting; Ritalin; mPFC; Neurotransmitter; Coularray detection

Gene expression of synaptosomal-associated protein 25 (SNAP-25) in the prefrontal cortex of the spontaneously hypertensive rat (SHR)

Dopamine is believed to play an important role in the etiology of attention-deficit/hyperactivity disorder (ADHD). In our previous study, we showed that gene expression of dopamine D4 receptor decreased in the spontaneously hypertensive rat (SHR) in the prefrontal cortex (PFC). In the present study, we explored the potential causes of dysfunction in the dopamine system in ADHD. It is the first time that neuronal activities in both juvenile SHR and WKY rats have been measured by functional MRI (fMRI). Our results showed that in PFC the Blood Oxygenation Level Dependent (BOLD) signal response in SHR was much higher than WKY under stressful situations. We tested the effects of acute and repeated administration of amphetamine on behavioral changes in SHR combined with the expression of the neuronal activity marker, c-fos, in the PFC. Meanwhile dopamine-related gene expression was measured in the PFC after repeated administration of amphetamine. We found that potential neuronal damage occurred through deficit of D2-like receptor protective functions in the PFC of the SHR. We also measured the expression of synaptosomal-associated protein 25 (SNAP-25) in SHR in PFC. The results showed decreased expression of SNAP-25 mRNA in the PFC of SHR; this defect disappeared after repeated injection of D-AMP.

Amplified respiratory–sympathetic coupling in the spontaneously hypertensive rat: does it contribute to hypertension?

Sympathetic nerve activity (SNA) is elevated in established hypertension. We tested the hypothesis that SNA is elevated in neonate and juvenile spontaneously hypertensive (SH) rats prior to the development of hypertension, and that this may be due to augmented respiratory-sympathetic coupling. Using the working heart-brainstem preparation, perfusion pressure, phrenic nerve activity and thoracic (T8) SNA were recorded in male SH rats and normotensive Wistar-Kyoto (WKY) rats at three ages: neonates (postnatal day 9-16), 3 weeks old and 5 weeks old. Perfusion pressure was higher in SH rats at all ages reflecting higher vascular resistance. The amplitude of respiratory-related bursts of SNA was greater in SH rats at all ages (P < 0.05). This was reflected in larger Traube-Hering pressure waves in SH rats (1.4 +/- 0.8 versus 9.8 +/- 1.5 mmHg WKY versus SH rat, 5 weeks old, n = 5 per group, P < 0.01). Recovery from hypocapnic-induced apnoea and reinstatement of Traube-Hering waves produced a significantly greater increase in perfusion pressure in SH rats (P < 0.05). Differences in respiratory-sympathetic coupling in the SH rat were not secondary to changes in central or peripheral chemoreflex sensitivity, nor were they related to altered arterial baroreflex function. We have shown that increased SNA is already present in SH rats in early postnatal life as revealed by augmented respiratory modulation of SNA. This is reflected in an increased magnitude of Traube-Hering waves resulting in elevated perfusion pressure in the SH rat. We suggest that the amplified respiratory-related bursts of SNA seen in the neonate and juvenile SH rat may be causal in the development of their hypertension.

High dietary alpha tocopherol improves attention deficit/hyperactivity (ADHD)‐like behavior in juvenile spontaneously hypertensive rats (SHR)

The juvenile SHR, a widely used animal model for ADHD, was previously shown to have low levels of alpha tocopherol (AT) in plasma and brain compared to Sprague-Dawley (SD) rats on the same diet. In this study we asked if AT levels are related to the behavior abnormalities associated with ADHD (inattention, hyperactivity, and impulsivity), using measures previously shown to distinguish these behaviors. Because AT might affect behavior by protection of important brain PUFAs such as arachidonic and docosahexaenoic acids (AA and DHA), fatty acid analysis was also done. Three groups of SHR (n=8 each) were fed synthetic AIN93 diets containing, respectively, 0, 50 and 250 U/kg AT acetate, and 8 SD rats were given the 50 U/kg AT diet. The diets were fed ad libitum from the age of 4 weeks. Behavior testing was done from 8 weeks to 12 weeks in conditioning cages equipped with infrared monitors. The animals were killed at 12 weeks. AT concentrations in plasma of SHRs increased with increasing dietary AT (1.47 ± 0.64, 56.3 ± 6.39, 69.4 ± 12.8 μM), approaching that of SDs (71.0 ± 12.1 μM). AT concentrations in brain followed a similar pattern (5.49 ± 0.54, 14.5 ± 0.69, 17.2 ± 1.06 μg/g compared to 18.6 ± 2.00 μg/g). Total n-3 PUFA in SHR plasma and the AA + DHA sum in SHR brain also increased in response to dietary AT. SHR behavior showed significant effects of dietary treatment. Measures of learning (attention), inhibition, and impulsivity improved with increasing AT. Total activity was not reduced by AT. Replenishment of AT in the SHR can improve behavior. Supported by the Purdue Research Foundation.

Model of Attention Deficit Hyperactivity Disorder: Five‐Trial, Repeated Acquisition Inhibitory Avoidance in Spontaneously Hypertensive Rat Pups

There is no one definitive animal model or test for ADHD. However, behavioral symptoms of this disorder, such as inattentiveness, impulsivity, rate of learning, and hyperactivity, can be evaluated in animals. Unfortunately, many of the cognitive tests used to assess attention and impulsivity require considerable equipment, and financial and time resources. Additionally, many of the genetic animal models are difficult to obtain due to cost, intellectual property issues, or difficulties in breeding sufficient numbers. The spontaneously hypertensive rat (SHR) is relatively easily obtained and exhibits many behavioral abnormalities analogous to those observed in ADHD. This unit describes a cost-effective and relatively simple approach to modeling ADHD preclinically, by training juvenile SHR pups in a five-trial repeated acquisition inhibitory avoidance model with components of attention and response inhibition. The model is suited to identifying compounds with stimulant and nonstimulant mechanisms of action, although the latter has yet to be verified conclusively in the clinic.

In vivo hypothalamic release and synthesis of catecholamines in spontaneously hypertensive rats.

Juvenile spontaneously hypertensive rats (SHR) have higher plasma levels of catechols and markedly larger catechol responses to yohimbine than do normotensive Wistar-Kyoto rats, indicating increased sympathoadrenal outflow and increased alpha 2-adrenergic receptor-mediated restraint of peripheral catecholamine release during hypertension development in SHR. Yohimbine-induced catecholamine release and metabolism in the posterolateral hypothalamus of the brain were assessed in juvenile (6 to 7 weeks) and adult (15 to 16 weeks) SHR and Wistar-Kyoto rats. In vivo microdialysis was used to obtain samples for measurements of norepinephrine, dihydroxyphenylglycol, methoxyhydroxyphenylglycol, and dihydroxyphenylacetic acid in conscious animals before and after yohimbine injection (1 mg/kg IV) beginning 24 hours after probe implantation. Catecholamine synthesis was examined from elevations of 3,4-dihydroxyphenylalanine levels after probe perfusion with NSD-1015, an inhibitor of L-aromatic acid decarboxylase. In adults, SHR had higher dialysate norepinephrine (277 +/- 38 versus 181 +/- 35 pg/mL), dihydroxyphenylglycol (3260 +/- 509 versus 2231 +/- 201 pg/mL), methoxyhydroxyphenylglycol (2659 +/- 369 versus 1890 +/- 144 pg/mL), and dihydroxyphenylacetic acid (46,312 +/- 5512 versus 13,187 +/- 1963 pg/mL) levels and markedly larger increases in 3,4-dihydroxyphenylalanine levels after NSD-1015 than Wistar-Kyoto rats. In juveniles, SHR had larger proportionate increments in microdialysate norepinephrine levels after yohimbine than Wistar-Kyoto rats (85% versus 25%). Although juvenile SHR and Wistar-Kyoto rats had similar NSD-1015-elicited increments in 3,4-dihydroxyphenylalanine levels, systemic yohimbine enhanced the NSD-1015-elicited 3,4-dihydroxyphenylalanine elevations in juvenile SHR but not in Wistar-Kyoto rats. These findings suggest augmented norepinephrine release and catecholamine synthesis in the posterolateral hypothalamus of adult SHR and augmented alpha 2-adrenergic receptor restraint of both norepinephrine release and catecholamine synthesis in juvenile SHR.

Endothelin-1 binding sites and inositol-monophosphate formation in kidney medulla from adult and juvenile SHR and WKY rats

High resolution autoradiography at the cellular level localized endothelin-1 binding sites to the collecting ducts in the rat renal papilla. These receptors were functionally coupled to the phosphatidyl-inositol system since endothelin-1 stimulated the accumulation of IP(1) in a concentration-dependent manner in cross chopped slices from renal papillae. The concentration-effect curves in 4 week old normotensive Wistar-Kyoto rats (WKY) lay to the right of curves from 4 week old and adult spontaneously hypertensive rats (SHR) and adult WKY (20-24 week old) animals; the EC(50) values in the 4 week old animals were 17 +/- 5 and 8 +/- 1 nM (P < 0.05, n = 5; mean +/- SE) for the normotensive and hypertensive animals, respectively. Autoradiographic studies showed that the density and distribution of binding sites for [(125)I]endothelin-1 in the kidneys did not differ between the groups; receptor densities in the renal papillae were 461 +/- 37 (fmol/mg protein) in the 4 week old WKY, and 443 +/- 27 (fmol/mg protein) in the 4 week old SHR. The plasma levels of immunoreactive endothelin-1 were also similar between groups; 4 week old SHR (39 +/- 3 pg/ml) and 4 week old WKY (36 +/- 1 pg/ml). The increased response to endothelin-1 may be related to the development of the hypertensive state in the SHR.