Amiloride Inhibition of ␥-Aminobutyric Acid A Receptors Depends upon the ␣ Subunit Subtype

ABSTRACT ␥-Aminobutyric acid A (GABA A ) receptors (GABARs) are responsible for most fast inhibitory neurotransmission in the mammalian brain. The GABARs contain several allosteric modulatory sites, many of which are useful clinically. The activity of most of these modulators depends upon the subunit composition of the receptor. The diuretic amiloride was previously reported to inhibit GABARs in frog sensory neurons. We measured its effects on recombinant GABARs to determine its mechanism of action at mammalian receptors and to examine the effect of subunit composition. Amiloride acted primarily as a competitive antagonist, reducing the sensitivity of the receptor to GABA without affecting the maximal current amplitude. Receptors containing an ␣6 subunit were about 10-fold more sensitive to amiloride than those containing other ␣ subunits. In contrast, the identity of the ␤ or ␥ subtype had little effect on amiloride sensitivity. Although several other modulators have specific effects at ␣6-containing receptors, amiloride is the first inhibitor to be reported with no additional dependence on the identity of the ␤ or ␥ subunit. Therefore, it probably represents a unique modulatory site on the GABAR, which could be useful for developing drugs targeting these receptors. The selective activity of amiloride could also be helpful for isolating the contribution of receptors composed of ␣6 subtypes in heterogeneous native GABAR populations

Clobazam as an adjunctive therapy in treating seizures associated with Lennox–Gastaut syndrome

Lennox–Gastaut syndrome (LGS) is a devastating childhood epilepsy syndrome characterized by the occurrence of multiple types of seizures and cognitive decline. Most children suffer from frequent seizures that are refractory to current medical management. Recent clinical trials have suggested that addition of clobazam may improve the clinical outcome for some LGS patients. Although clobazam has been available for over five decades, it has only recently been approved by the US Food and Drug Administration for this indication. As a 1,5-benzodiazepine, clobazam is structurally related to the widely used 1,4-benzodiazepines, which include diazepam. Clobazam has been shown to modulate GABAergic neurotransmission by positive allosteric modulation of GABAA receptors, and to increase expression of transporters for both GABA and glutamate. The active metabolite n-desmethylclobazam (norclobazam) also modulates GABAA receptors, and the relative importance of these two compounds in the clinical effectiveness of clobazam remains an open question. Clinical trials involving clobazam as an addon therapy in a variety of pediatric epilepsy populations have found a significant improvement in seizure control. In patients with LGS, clobazam may have greatest efficacy for drop seizures. Longstanding clinical experience suggests that clobazam is a safe and well tolerated antiepileptic drug with infrequent and mild adverse effects. These results suggest that adjunctive treatment with clobazam may be a reasonable option for LGS patients, particularly those who are treatment-resistant

Trajectories Of Zooplankton Recovery In The Little Rock Lake Whole‐Lake Acidification Experiment

Understanding the factors that affect biological recovery from environmental stressors such as acidification is an important challenge in ecology. Here we report on zooplankton community recovery following the experimental acidification of Little Rock Lake, Wisconsin, USA. One decade following cessation of acid additions to the northern basin of Little Rock Lake (LRL), recovery of the zooplankton community was complete. Approximately 40% of zooplankton species in the lake exhibited a recovery lag in which biological recovery to reference basin levels was delayed by 1–6 yr after pH recovered to the level at which the species originally responded. Delays in recovery such as those we observed in LRL may be attributable to “biological resistance” wherein establishment of viable populations of key acid-sensitive species following water quality improvements is prevented by other components of the community that thrived during acidification. Indeed, we observed that the recovery of species that thrived during acidification tended to precede recovery of species that declined during acidification. In addition, correspondence analysis indicated that the zooplankton community followed different pathways during acidification and recovery, suggesting that there is substantial hysteresis in zooplankton recovery from acidification. By providing an example of a relatively rapid recovery from short-term acidification, zooplankton community recovery from experimental acidification in LRL generally reinforces the positive outlook for recovery reported for other acidified lakes

Fluoxetine Increases GABA A Receptor Activity through a Novel Modulatory Site

ABSTRACT Fluoxetine is a selective serotonin reuptake inhibitor used widely in the treatment of depression. In contrast to the proconvulsant effect of many antidepressants, fluoxetine has anticonvulsant activity. This property may be due in part to positive modulation of the GABA A receptors (GABARs), which mediate most fast inhibitory neurotransmission in the mammalian brain. We examined the effect of fluoxetine on the activity of recombinant GABARs transiently expressed in mammalian cells. Fluoxetine increased the response of the receptor to submaximal GABA concentrations but did not alter the maximum current amplitude. Sensitivity did not depend upon the ␤-or ␥-subtype composition of the receptor when coexpressed with the ␣ 1 subunit. Among the six ␣ subtypes, only the ␣ 5 subunit conferred reduced sensitivity to fluoxetine. The metabolite norfluoxetine was even more potent than fluoxetine. Mutations at residues in the ␣ 5 subunit that alter its sensitivity to zinc or selective benzodiazepine derivatives did not affect potentiation by fluoxetine. This suggests that fluoxetine acts through a novel modulatory site on the GABAR. The direct positive modulation of GABARs by fluoxetine may be a factor in its anticonvulsant activity

Spontaneous and �-aminobutyric acid (GABA)-activated GABA A receptor channels formed by � subunit-containing isoforms

ABSTRACT A new ␥-aminobutyric acid (GABA) A receptor (GABAR) subunit class, ⑀, has recently been cloned and shown to form functional channels when coexpressed with both ␣ and ␤ subunits. We report that the combination of ␣1␤3⑀ subunit subtypes expressed in L929 cells produced functional chloride ion channels that were both spontaneously active and gated by the application of extracellular GABA. When cells were voltage-clamped at -75 mV in the whole-cell configuration, holding currents of 50 to 300 pA associated with increased noise were consistently recorded. The application of pentobarbital and loreclezole, which increase GABAR currents, increased the holding current, whereas the application of zinc and picrotoxin, which reduce GABAR currents, reduced the holding current in a concentration-dependent manner. Coexpression of ␣1␤3␥2L, ␣1␤3␦, ␣1⑀, ␤3⑀, ␣1␤3, or ⑀ subtypes did not produce holding currents that were sensitive to picrotoxin (30 M). Cells expressing ␣1 ␤3⑀ subtypes had concentration-dependent GABAR currents that were potentiated by pentobarbital, loreclezole, and lanthanum and inhibited by zinc and furosemide. Spontaneous and GABAR single-channel currents from ␣1␤3⑀ receptors had single-channel conductances of ϳ24 pS. The biophysical properties and the effects of allosteric modulators were similar for spontaneous and evoked GABAR currents, suggesting that a single GABAR isoform was responsible for both currents. These data extend the pharmacological characterization of ⑀-containing GABARs and demonstrate that incorporation of the ⑀ subunit permits spontaneous channel gating while preserving the structural information necessary for GABA sensitivity

A theory-based approach to understanding condom errors and problems reported by men attending an STI clinic

The official published version can be accessed from the link below - Copyright @ 2008 Springer VerlagWe employed the information–motivation–behavioral skills (IMB) model to guide an investigation of correlates for correct condom use among 278 adult (18–35 years old) male clients attending a sexually transmitted infection (STI) clinic. An anonymous questionnaire aided by a CD-recording of the questions was administered. Linear Structural Relations Program was used to conduct path analyses of the hypothesized IMB model. Parameter estimates showed that while information did not directly affect behavioral skills, it did have a direct (negative) effect on condom use errors. Motivation had a significant direct (positive) effect on behavioral skills and a significant indirect (positive) effect on condom use errors through behavioral skills. Behavioral skills had a direct (negative) effect on condom use errors. Among men attending a public STI clinic, these findings suggest brief, clinic-based, safer sex programs for men who have sex with women should incorporate activities to convey correct condom use information, instill motivation to use condoms correctly, and directly enhance men’s behavioral skills for correct use of condoms

Single channel properties of recombinant GABA A receptors containing Γ2 or Δ subtypes expressed with Α1 and Β3 subtypes in mouse L929 Cells

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65956/1/j.1469-7793.1997.283bb.x.pd

Teaching Mathematics with Technology: TPACK and Effective Teaching Practices

This paper examines how 17 secondary mathematics teacher candidates (TCs) in four university teacher preparation programs implemented technology in their classrooms to teach for conceptual understanding in online, hybrid, and face to face classes during COVID-19. Using the Professional Development: Research, Implementation, and Evaluation (PrimeD) framework, TCs, classroom mentor teachers, field experience supervisors, and university faculty formed a Networked Improvement Community (NIC) to discuss a commonly agreed upon problem of practice and a change idea to implement in the classroom. Through Plan-Do-Study-Act cycles, participants documented their improvement efforts and refinements to the change idea and then reported back to the NIC at the subsequent monthly meeting. The Technology Pedagogical Content Knowledge framework (TPACK) and the TPACK levels rubric were used to examine how teacher candidates implemented technology for Mathematics conceptual understanding. The Mathematics Classroom Observation Protocol for Practices (MCOP2) was used to further examine how effective mathematics teaching practices (e.g., student engagement) were implemented by TCs. MCOP2 results indicated that TCs increased their use of effective mathematics teaching practices. However, growth in TPACK was not significant. A relationship between TPACK and MCOP2 was not evident, indicating a potential need for explicit focus on using technology for mathematics conceptual understanding

Rental Housing Assistance for the 21st Century

Current rental housing assistance programs are not designed to provide a safety net for people whose lives are volatile, or to encourage poor people to live in good locations. These failings can be corrected. HUD should establish a program of rental insurance-like mortgage insurance, but for renters. Low income housing assistance formulas should be revised to reward good neighborhood features, and punish bad