Supplement: Calcineurin and Inflammatory Signaling Processes in Aging and Alzheimer's Disease

NORRIS RF1-AG027297 SUPPLEMENT The Problem: As outlined in our Parent RF1, the completion of Aim 2 studies on the role of reactive astrocytes in cerebrovascular dysfunction in AD, VCID, and AD/VCID mouse models require intravital two photon microscopy. The Zeiss LSM 880— which was expected to be available for the proposed studies— was shut down at the early part of the COVID pandemic (Spring 2020) and subsequently developed several maintenance issues resulting in extensive downtime in the Spring-Summer of 2021. After initiation of our RF1, a second two photon system (a Scientifica two photon microscope) was established in the Sanders-Brown Center on Aging (SBCoA) and was fully operational in the latter part of 2019. Note, because of the Zeiss system, we never intended to use the SBCoA microscope to complete studies on our RF1. However, during the COVID shutdown— when the Zeiss was unavailable— we were able to use the Scientifica system extensively, which led to significant progress on Aim 2 studies. However, as university operations picked up again after the COVID shutdown, our time on the Scientifica has dramatically decreased due to increased usage by other NIH funded projects in the SBCoA. The combined issues across both microscopes has created a usage bottleneck that jeopardizes the completion of Aim 2 by the end of the funding period (March 2023). Need: Increased two photon imaging capacity. Solution: Convert a recently purchased Scientifica patch clamp system— housed in the Norris laboratory and dedicated to Norris lab research— into a fully operational two photon instrument. Last year, the Norris lab purchased a Scientifica patch clamp electrophysiology station through a retention package at UK. While the system is presently used for epifluorescent microscopy and patch clamp applications, it was future-proofed for the eventual accommodation of two photon imaging capabilities, pending funding. To convert this system to a fully operational two photon system we need a high-performance laser and Scientifica dual scan path launch optics. Funds from this supplement award will cover these costs.