skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

Explore Research Products in the PAR
It may take a few hours for recently added research products to appear in PAR search results.


  • Home
  • Search Results
  • Page 1 of 1

Search for: All records

Creators/Authors contains: "Simpson, Hailey"

Note: When clicking on a Digital Object Identifier (DOI) number, you will be taken to an external site maintained by the publisher. Some full text articles may not yet be available without a charge during the embargo (administrative interval).
What is a DOI Number?

Some links on this page may take you to non-federal websites. Their policies may differ from this site.

  1. ; ; ; (, Frontiers in Cellular Neuroscience)
    Postsynaptic cytosolic Cl − concentration determines whether GABAergic and glycinergic synapses are inhibitory or excitatory. We have shown that nitric oxide (NO) initiates the release of Cl − from acidic internal stores into the cytosol of retinal amacrine cells (ACs) thereby elevating cytosolic Cl − . In addition, we found that cystic fibrosis transmembrane conductance regulator (CFTR) expression and Ca 2+ elevations are necessary for the transient effects of NO on cytosolic Cl − levels, but the mechanism remains to be elucidated. Here, we investigated the involvement of TMEM16A as a possible link between Ca 2+ elevations and cytosolic Cl − release. TMEM16A is a Ca 2+ -activated Cl − channel that is functionally coupled with CFTR in epithelia. Both proteins are also expressed in neurons. Based on this and its Ca 2+ dependence, we test the hypothesis that TMEM16A participates in the NO-dependent elevation in cytosolic Cl − in ACs. Chick retina ACs express TMEM16A as shown by Western blot analysis, single-cell PCR, and immunocytochemistry. Electrophysiology experiments demonstrate that TMEM16A functions in amacrine cells. Pharmacological inhibition of TMEM16A with T16inh-AO1 reduces the NO-dependent Cl − release as indicated by the diminished shift in the reversal potential of GABA A receptor-mediated currents. We confirmed the involvement of TMEM16A in the NO-dependent Cl − release using CRISPR/Cas9 knockdown of TMEM16A. Two different modalities targeting the gene for TMEM16A ( ANO1 ) were tested in retinal amacrine cells: an all-in-one plasmid vector and crRNA/tracrRNA/Cas9 ribonucleoprotein. The all-in-one CRISPR/Cas9 modality did not change the expression of TMEM16A protein and produced no change in the response to NO. However, TMEM16A-specific crRNA/tracrRNA/Cas9 ribonucleoprotein effectively reduces both TMEM16A protein levels and the NO-dependent shift in the reversal potential of GABA-gated currents. These results show that TMEM16A plays a role in the NO-dependent Cl − release from retinal ACs. 
    more » « less
    Full Text Available