• Skip to primary navigation
  • Skip to main content
SRI logo
  • About
    • Press room
    • Our history
  • Expertise
    • Advanced imaging systems
    • Artificial intelligence
    • Biomedical R&D services
    • Biomedical sciences
    • Computer vision
    • Cyber & formal methods
    • Education and learning
    • Innovation strategy and policy
    • National security
    • Ocean & space
    • Quantum
    • Robotics, sensors & devices
    • Speech & natural language
    • Video test & measurement
  • Ventures
  • NSIC
  • Careers
  • Contact
  • 日本支社
Search
Close
Biomedical sciences publications August 1, 2014

Cortical nNOS/NK1 Neurons Are Regulated by Cholinergic Inputs

Thomas Kilduff

Citation

Copy to clipboard


Williams, R. H., Vasquez-DeRose, J., Nguyen, A., & Kilduff, T. S. (2014, 15-19 November). Cortical nNOS/NK1 neurons are regulated by cholinergic inputs. Paper presented at Neuroscience 2014, Washington, DC.

Abstract

Cortical GABAergic interneurons shown to co-express neuronal nitric oxide synthase (nNOS) and the neurokinin-1 (NK1) receptor are activated during sleep (Dittrich et al., 2012, Front. Neural Cir). These neurons have been proposed to play a role in sleep homeostasis (Gerashchenko et al., 2008, PNAS)as the proportion of this population expressing cFOS correlates with homeostatic sleep drive (Morairty et al 2013, PNAS). As these cells appear to “”sense”” sleep need, we investigated whether cholinergic inputs affect the electrophysiological properties of cortical nNOS/NK1 neurons. We prepared coronal mouse brain slices (250µm thick) for whole-cell patch-clamp recording in both voltage clamp and current clamp modes. After a brief application of the fluorescent NK1 agonist tetramethylrhodamine (TMR), cortical nNOS/NK1 neurons were readily identifiable. Following a 60min wash-out period to eliminate any residual TMR-mediated response, carbachol (CCh, a cholinergic mimetic) was applied. All cells tested responded to CCh (50µM; n=34). CCh had biphasic effects: an initial membrane hyperpolarization (-2.24±0.57mV, n=6) and outward current (+3.74±0.66pA, n=12) was followed by a large membrane depolarization (+9.18±1.87mV, n=8) and inward current (-5.59±2.22pA, n=12). In addition, CCh predominantly increased spontaneous excitatory postsynaptic currents (sEPSCs; +805.8±62.68%, n=15). In the presence of tetrodotoxin (TTX), both outward (+3.18±0.74pA, n=9), and inward (-11.96±1.88pA, n=15) currents remained, indicating responses mediated by postsynaptic receptors. We tested whether the excitatory response mediated by CCh was attributable to activation of the Gq-coupled muscarinic type 1 receptors (M1R). Application of CCh in the presence of the M1R antagonist VU0235535 (5μM) reduced the membrane depolarization by 47.9±16.9% and inward current by 32.0±17.7% in 3/5 cells tested. The 2 remaining cells showed no change in their excitatory response but an increase in the outward current (+5.76±2.11pA, n=2). In contrast to sEPSCs, CCh decreased miniature EPSCs (mEPSCs) by -45.6±2.2% (n=6). The presence of VU0235535 did little to alter the reduction in mEPSCs seen following CCh application (after vs baseline: -53.1±2.8%, n=4), suggesting that M1Rs are largely postsynaptic. In summary, CCh has a network effect on excitability within the cortex, as both presynaptic and postsynaptic effects result in an oscillating pattern of nNOS/NK1 neuron excitability. When all inputs are blocked, the predominant response is excitation. We are currently attempting to determine the source of cholinergic inputs to cortical nNOS/NK1 neurons.

Keywords: acetylcholine, arousal, cortex, muscarinic, nicotinic, nitric oxide, sleep, wakefulness.

↓ View online

Share this
Career call to action image

Work with us

Search jobs

How can we help?

Once you hit send…

We’ll match your inquiry to the person who can best help you.

Expect a response within 48 hours.

Our work

Case studies

Publications

Timeline of innovation

Areas of expertise

Institute

Leadership

Press room

Media inquiries

Compliance

Careers

Job listings

Contact

SRI Ventures

Our locations

Headquarters

333 Ravenswood Ave
Menlo Park, CA 94025 USA

+1 (650) 859-2000

Subscribe to our newsletter


日本支社
SRI International
  • Contact us
  • Privacy Policy
  • Cookies
  • DMCA
  • Copyright © 2023 SRI International
Manage Cookie Consent
To provide the best experiences, we use technologies like cookies to store and/or access device information. Consenting to these technologies will allow us to process data such as browsing behavior or unique IDs on this site. Not consenting or withdrawing consent, may adversely affect certain features and functions.
Functional Always active
The technical storage or access is strictly necessary for the legitimate purpose of enabling the use of a specific service explicitly requested by the subscriber or user, or for the sole purpose of carrying out the transmission of a communication over an electronic communications network.
Preferences
The technical storage or access is necessary for the legitimate purpose of storing preferences that are not requested by the subscriber or user.
Statistics
The technical storage or access that is used exclusively for statistical purposes. The technical storage or access that is used exclusively for anonymous statistical purposes. Without a subpoena, voluntary compliance on the part of your Internet Service Provider, or additional records from a third party, information stored or retrieved for this purpose alone cannot usually be used to identify you.
Marketing
The technical storage or access is required to create user profiles to send advertising, or to track the user on a website or across several websites for similar marketing purposes.
Manage options Manage services Manage {vendor_count} vendors Read more about these purposes
View preferences
{title} {title} {title}