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Biology subjects

Hu, J. Y.

Publications and source records attributed to Hu, J. Y..

4 recordsLinked to original sources

Rapid spatial cognition in mice, with and without neocortex and hippocampus

Rapid learning, memory, and generalization are often attributed to circuits of the neocortex and hippocampus, but their specific role remains unclear. To examine these cognitive abilities together in individual mice, we observed mice navigating the Manhattan Maze, a reconfigurable 3D labyrinth. Naive wildtype mice improved within two rewards, approached efficient paths within about 20 rewards, retained a 9-turn route overnight, and learned faster in new configurations. Much of the few-shot improvement follows from a rule-based forward bias that emerged even before any reward. In a maze with loops, where that rule is less useful, mice learned within a few rewards to prefer one bottleneck corridor far from the reward location, while choices elsewhere stayed flexible. To begin linking these different components of learning to brain function, we presented the same task to mutant mice that lack the hippocampus and most of the neocortex. They were impaired during initial exploration, where repetitive scanning made them about 3-fold slower to obtain the first few rewards. Past that stage, learning, retention over weeks to months, and generalization to new mazes were largely preserved. Learning this fast is hard to reconcile with trial and error, in which value propagates backwards from the reward. A neuromorphic circuit model instead accounts for the bottleneck choice: it builds a map of the environment without reward, and needs neither cortical nor hippocampal circuit motifs. Therefore, learning is possible without the involvement of neocortex and hippocampus. Structure learned before the first reward, rather than the reward itself, may be what makes few-shot learning possible.

neuroscience↗

A Far-Red FRET biosensor for AMPK enables multiplexed imaging of single-cell bioenergetic homeostasis

Metabolic homeostasis has been studied primarily at the tissue and organism level, identifying molecular control mechanisms such as the energy charge-sensing kinase AMPK. Feedback loops involving AMPK and other regulators align cellular ATP generation and consumption, determining energetic balance. Recent work has demonstrated surprising oscillatory dynamics in AMPK activity, revealing unidentified kinetic modulation in single-cell homeostatic behaviour. However, probing the kinetic mechanisms of intracellular feedback requires simultaneous observation of multiple energetic parameters, and such experiments are precluded by the shared wavelength band occupied by most metabolic biosensors. We have overcome this obstacle by constructing a red-shifted FRET-based AMPK activity biosensor, RAMPKAR2, that is comparable to existing FRET-based AMPK activity biosensors. Multiplexed imaging of RAMPKAR2 with PercevalHR, which detects ATP/ADP ratio, confirmed that the kinetics of AMPK activity and ATP/ADP ratio are tightly coupled, with a lag of less than 6 minutes at the single-cell level. Pairing of RAMPKAR with HYlight, which detects the glycolytic intermediate fructose 1,6-bisphosphate (FBP), revealed that glycolytic activity co-oscillates with AMPK, shifted by [~]1.5 hours, and that these oscillations are suppressed by sustained AMPK activity. Together these data advance a model in which temporally offset increases in glycolytic ATP supply and AMPK deactivation contribute to single-cell oscillations.

systems biology↗

Two Novel Red-FRET ERK Biosensors in the 670-720nm Range.

Cell fate decisions are regulated by intricate signaling networks, with Extracellular signal-Regulated Kinase (ERK) being a central regulator. However, ERK is rarely the only signaling pathway involved, creating a need to study multiple signaling pathways simultaneously at the single-cell level. Many existing fluorescent biosensors for ERK and other pathways have significant spectral overlap, limiting their ability to be multiplexed. To address this limitation, we developed two novel red-FRET ERK biosensors, REKAR67 and REKAR76, which operate in the 670-720 nm range using miRFP670nano3 and miRFP720. REKAR67 and REKAR76 differ in fluorophore position, which impacts biosensor characteristics; REKAR67 displayed a higher dynamic range but greater signal variance than REKAR76. Mixed populations of REKAR67 or REKAR76 displayed similar Signal-to-Noise ratio (SNR), but in clonal cell populations, REKAR76 had a significantly higher SNR. Overall, our red-FRET ERK biosensors were highly consistent with existing ERK FRET biosensors and in reporting ERK activity and are spectrally compatible with CFP/YFP FRET and cpGFP -based biosensors. Both REKAR biosensors expand the available methods for measuring single-cell ERK activity.

cell biology↗

Dynamic regulation of the oxidative stress response by the E3 ligase TRIP12

The oxidative stress response is centered on the transcription factor NRF2 and protects cells from reactive oxygen species (ROS). While ROS inhibit the E3 ligase CUL3KEAP1 to stabilize NRF2 and elicit antioxidant gene expression, cells recovering from stress must rapidly reactivate CUL3KEAP1 to prevent reductive stress and oxeiptosis-dependent cell death. How cells restore efficient NRF2-degradation upon ROS clearance remains poorly understood. Here, we identify TRIP12, an E3 ligase dysregulated in Clark-Baraitser Syndrome and Parkinsons Disease, as a component of the oxidative stress response. TRIP12 is a ubiquitin chain elongation factor that cooperates with CUL3KEAP1 to ensure robust NRF2 degradation. In this manner, TRIP12 accelerates stress response silencing as ROS are being cleared, but limits NRF2 activation during stress. The need for dynamic control of NRF2-degradation therefore comes at the cost of diminished stress signaling, suggesting that TRIP12 inhibition could be used to treat degenerative pathologies characterized by ROS accumulation.

biochemistry↗