Unbiased high-content screening reveals Aβ- and tau-independent synaptotoxic activities in human brain homogenates from Alzheimer’s patients and high-pathology controls

Alzheimer’s disease (AD) is tightly correlated with synapse loss in vulnerable brain regions. It is assumed that specific molecular entities such as Aβ and tau cause synapse loss in AD, yet unbiased screens for synaptotoxic activities have not been performed. Here, we performed size exclusion chromatography on soluble human brain homogenates from AD cases, high pathology non-demented controls, and low pathology age-matched controls using our novel high content primary cultured neuron-based screening assay. Both presynaptic and postsynaptic toxicities were elevated in homogenates from AD cases and high pathology non-demented controls to a similar extent, with more modest synaptotoxic activities in homogenates from low pathology normal controls. Surprisingly, synaptotoxic activities were found in size fractions peaking between the 17–44 kDa size standards that did not match well with Aβ and tau immunoreactive species in these homogenates. The fractions containing previously identified high molecular weight soluble amyloid beta aggregates/”oligomers” were non-toxic in this assay. Furthermore, immunodepletion of Aβ and tau did not reduce synaptotoxic activity. This result contrasts with previous findings involving the same methods applied to 3xTg-AD mouse brain extracts. The nature of the synaptotoxic species has not been identified. Overall, our data indicates one or more potential Aβ and tau independent synaptotoxic activities in human AD brain homogenates. This result aligns well with the key role of synaptic loss in the early cognitive decline and may provide new insight into AD pathophysiology.

Cytoskeletal Filaments Deep Inside a Neuron Are Not Silent: They Regulate the Precise Timing of Nerve Spikes Using a Pair of Vortices

Hodgkin and Huxley showed that even if the filaments are dissolved, a neuron’s membrane alone can generate and transmit the nerve spike. Regulating the time gap between spikes is key to the brain’s cognitive function; however, the time modulation mechanism is still a mystery. By inserting a coaxial probe deep inside a neuron, we repeatedly show that the filaments transmit electromagnetic signals of ~200 μs before an ionic nerve spike sets in. To understand its origin, here, we mapped the electromagnetic vortex produced by a filamentary bundle deep inside a neuron, regulating the nerve spike’s electrical-ionic vortex. We used monochromatic polarized light to measure the transmitted signals beating from the internal components of a cultured neuron. A nerve spike is a 3D ring of the electric field encompassing the perimeter of a neural branch. Several such vortices flow sequentially to keep precise timing for the brain’s cognition. The filaments hold millisecond order time gaps between membrane spikes with microsecond order signaling of electromagnetic vortices. Dielectric resonance images revealed that ordered filaments inside neural branches instruct the ordered grid-like network of actin–beta-spectrin just below the membrane. That layer builds a pair of electric field vortices, which coherently activates all ion-channels in a circular area of the membrane lipid bilayer when a nerve spike propagates. When biomaterials vibrate resonantly with microwave and radio-wave, simultaneous quantum optics capture ultra-fast events in a non-demolition mode, revealing multiple correlated time-domain operations beyond the Hodgkin–Huxley paradigm. Neuron holograms pave the way to understanding the filamentary circuits of a neural network in addition to membrane circuits.