Enteric Glia Modulate Macrophage Phenotype and Visceral Sensitivity following Inflammation.

Mechanisms resulting in abdominal pain include altered neuro-immune interactions in the gastrointestinal tract, but the signaling processes that link immune activation with visceral hypersensitivity are unresolved. We hypothesized that enteric glia link the neural and immune systems of the gut and that communication between enteric glia and immune cells modulates the development of visceral hypersensitivity. To this end, we manipulated a major mechanism of glial intercellular communication that requires connexin-43 and assessed the effects on acute and chronic inflammation, visceral hypersensitivity, and immune responses. Deleting connexin-43 in glia protected against the development of visceral hypersensitivity following chronic colitis. Mechanistically, the protective effects of glial manipulation were mediated by disrupting the glial-mediated activation of macrophages through the macrophage colony-stimulating factor. Collectively, our data identified enteric glia as a critical link between gastrointestinal neural and immune systems that could be harnessed by therapies to ameliorate abdominal pain.

Ammonia modifies enteric neuromuscular transmission through glial γ-aminobutyric acid signaling.

Impaired gut motility may contribute, at least in part, to the development of systemic hyperammonemia and systemic neurological disorders in inherited metabolic disorders, or in severe liver and renal disease. It is not known whether enteric neurotransmission regulates intestinal luminal and hence systemic ammonia levels by induced changes in motility. Here, we propose and test the hypothesis that ammonia acts through specific enteric circuits to influence gut motility. We tested our hypothesis by recording the effects of ammonia on neuromuscular transmission in tissue samples from mice, pigs, and humans and investigated specific mechanisms using novel mutant mice, selective drugs, cellular imaging, and enzyme-linked immunosorbent assays. Exogenous ammonia increased neurogenic contractions and decreased neurogenic relaxations in segments of mouse, pig, and human intestine. Enteric glial cells responded to ammonia with intracellular Ca2+ responses. Inhibition of glutamine synthetase and the deletion of glial connexin-43 channels in hGFAP::CreERT2+/-/connexin43f/f mice potentiated the effects of ammonia on neuromuscular transmission. The effects of ammonia on neuromuscular transmission were blocked by GABAA receptor antagonists, and ammonia drove substantive GABA release as did the selective pharmacological activation of enteric glia in GFAP::hM3Dq transgenic mice. We propose a novel mechanism whereby local ammonia is operational through GABAergic glial signaling to influence enteric neuromuscular circuits that regulate intestinal motility. Therapeutic manipulation of these mechanisms may benefit a number of neurological, hepatic, and renal disorders manifesting hyperammonemia.NEW & NOTEWORTHY We propose that local circuits in the enteric nervous system sense and regulate intestinal ammonia. We show that ammonia modifies enteric neuromuscular transmission to increase motility in human, pig, and mouse intestine model systems. The mechanisms underlying the effects of ammonia on enteric neurotransmission include GABAergic pathways that are regulated by enteric glial cells. Our new data suggest that myenteric glial cells sense local ammonia and directly modify neurotransmission by releasing GABA.

Enteric glia mediate neuron death in colitis through purinergic pathways that require connexin-43 and nitric oxide.

BACKGROUND AND AIMS: The concept of enteric glia as regulators of intestinal homeostasis is slowly gaining acceptance as a central concept in neurogastroenterology. Yet how glia contribute to intestinal disease is still poorly understood. Purines generated during inflammation drive enteric neuron death by activating neuronal P2X7 purine receptors (P2X7R), triggering ATP release via neuronal pannexin-1 channels that subsequently recruits intracellular calcium ([Ca2+]i) responses in the surrounding enteric glia. We tested the hypothesis that the activation of enteric glia contributes to neuron death during inflammation. METHODS: We studied neuroinflammation in vivo using the 2,4-dinitrobenzenesulfonic acid model of colitis and in situ using whole-mount preparations of human and mouse intestine. Transgenic mice with a targeted deletion of glial connexin-43 (Cx43) [GFAP∷CreERT2+/-/Cx43f/f ] were used to specifically disrupt glial signaling pathways. Mice deficient in inducible nitric oxide (NO) synthase (iNOS-/-) were used to study NO production. Protein expression and oxidative stress were measured using immunohistochemistry and in situ Ca2+ and NO imaging were used to monitor glial [Ca2+]i and [NO]i. RESULTS: Purinergic activation of enteric glia drove [Ca2+]i responses and enteric neuron death through a Cx43-dependent mechanism. Neurotoxic Cx43 activity, driven by NO production from glial iNOS, was required for neuron death. Glial Cx43 opening liberated ATP and Cx43-dependent ATP release was potentiated by NO. CONCLUSIONS: Our results show that the activation of glial cells in the context of neuroinflammation kills enteric neurons. Mediators of inflammation that include ATP and NO activate neurotoxic pathways that converge on glial Cx43 hemichannels. The glial response to inflammatory mediators might contribute to the development of motility disorders.

Hypertension in Asian/Pacific Island Americans.

Asian/Pacific Islander Americans (APIAs) are the fastest growing population in the United States by percentage. Hypertension is common and increases cardiovascular risk to a great extent in this population. The medical problems of this group are being increasingly encountered by US physicians. Many gene mutations associated with hypertension are more common in Asians. The significance of these polymorphisms in the pathogenesis of hypertension in APIAs is unclear. The percentage of APIAs who are aware, treated, and controlled is small. There may be some differences in the responses to antihypertensive medications between APIAs and whites. The results of human studies on the effect of drinking of tea on blood pressure in different groups are conflicting. Cough associated with angiotensin-converting enzyme inhibitor therapy may be more common in APIAs than in whites. There is a need for more education of APIAs regarding hypertension and for more effective treatment of hypertension by the physicians caring for this population.

Experimental animal models of hypertension.

Hypertension (HTN) and cardiovascular disease are the most common causes of death in developed countries. The use of experimental animal models of HTN has provided valuable information regarding many aspects of HTN, including etiology, pathophysiology, complications, and treatment. Because the etiology of HTN is heterogeneous, many experimental animal models have been developed to mimic the many facets of human HTN. The choice of animal model will be determined by the research question, monetary limitations, and technical expertise. The categories of models of HTN are: renovascular, renal parenchymal, pharmacologically induced, environmentally induced, and genetic. There are considerable differences between HTN in animals and humans, including differences in homeostatic mechanisms and pathophysiology; therefore, a thorough understanding of the animal models and rigorous analysis is required before extrapolating the finding in animals to humans.

Preferential myosin heavy chain isoform B Expression may contribute to the faster velocity of contraction in veins versus arteries.

Smooth muscle myosin heavy chains occur in 2 isoforms, SMA (slow) and SMB (fast). We hypothesized that the SMB isoform is predominant in the faster-contracting rat vena cava compared to thoracic aorta. We compared the time to half maximal contraction in response to a maximal concentration of endothelin-1 (ET-1; 100 nM), potassium chloride (KCl; 100 mM) and norepinephrine (NE; 10 microM). The time to half maximal contraction was shorter in the vena cava compared to aorta (aorta: ET-1 = 235.8 +/- 13.8 s, KCl = 140.0 +/- 33.3 s, NE = 19.8 +/- 2.7 s; vena cava: ET-1 = 121.8 +/- 15.6 s, KCl = 49.5 +/- 6.7 s, NE = 9.0 +/- 3.3 s). Reverse-transcription polymerase chain reaction supported the greater expression of SMB in the vena cava compared to aorta. SMB was expressed to a greater extent than SMA in the vessel wall of the vena cava. Western analysis determined that expression of SMB, relative to total smooth muscle myosin heavy chains, was 12.5 +/- 4.9-fold higher in the vena cava compared to aorta, while SMA was 4.9 +/- 1.2-fold higher in the aorta than vena cava. Thus, the SMB isoform is the predominant form expressed in rat veins, providing one possible mechanism for the faster response of veins to vasoconstrictors.

Morphological and biochemical characterization of remodeling in aorta and vena cava of DOCA-salt hypertensive rats.

Arterial remodeling occurs in response to mechanical and neurohumoral stimuli. We hypothesized that veins, which are not exposed to higher pressures in hypertension, would demonstrate less active remodeling than arteries. We assessed remodeling with two standard measures of arterial remodeling: vessel morphometry and the expression/function of matrix metalloproteinases (MMPs). Thoracic aorta and vena cava from sham normotensive and DOCA-salt hypertensive rats (110 +/- 4 and 188 +/- 8 mmHg systolic blood pressure, respectively) were used. Wall thickness was increased in DOCA-salt vs. sham aorta (301 +/- 23 vs. 218 +/- 14 mum, P < 0.05), as was medial area, but neither measure was altered in the vena cava. The aorta and vena cava expressed the gelatinases MMP-2, MMP-9, transmembrane proteinase MT1-MMP, and tissue inhibitor of metalloproteinase-2 (TIMP-2). Immunohistochemically, MMP-2 localized to smooth muscle in the aorta and densely in endothelium/smooth muscle of the vena cava. Western and zymographic analyses verified that MMP-2 was active in all vessels and less active in the vena cava than aorta. In hypertension, MMP-2 expression and activity in the aorta were increased (59.1 +/- 3.7 and 74.5 +/- 6.1 units in sham and DOCA, respectively, P < 0.05); similar elevations were not observed in the vena cava. MMP-9 was weakly expressed in all vessels. MT1-MMP was expressed by the aorta and vena cava and elevated in the vena cava from DOCA-salt rats. TIMP-2 expression was significantly increased in the aorta of DOCA rats compared with sham but was barely detectable in the vena cava of sham or DOCA-salt hypertensive rats. These findings suggest that large veins may not undergo vascular remodeling in DOCA-salt hypertension.

Changes in rates of beta-blocker use in community hospital patients with acute myocardial infarction.

OBJECTIVE: To examine changes in the rate of beta-blocker (BB) use at admission, in hospital, and at discharge between 1994 and 1995 (MICH I) and 1997 (MICH II) in patients with acute myocardial infarction (AMI). DESIGN: Comparison of two prospectively enrolled cohorts. SETTING: Five mid-Michigan community hospitals. PATIENTS: We studied 287 MICH I patients and 121 MICH II patients with AMI who had no contraindications to BB use from cohorts of consecutively admitted cases of AMI (814 in MICH I; 500 in MICH II). RESULTS: Prescription of BBs to ideal patients with AMI increased in patients with previous history of myocardial infarction on arrival at the hospital (12.5% vs 36.0%; P= .01), in hospital (47.0% vs 76%; P < .01), and at discharge (34.0% vs 61.9%; P < .01). Neither race nor gender was a predictor of BB use. Younger age predicted BB prescription at discharge (odds ratio [OR], 2.07; 95% confidence interval [CI], 1.32 to 3.23). Later study cohort was the most important predictor of BB use in hospital (OR, 3.4; 95% CI, 2.09 to 5.25). CONCLUSION: BB use improved dramatically over the study period, but additional work is needed to improve use of BB after discharge and among elderly patients with AMI.