Browning, P. G. F. | bioRxiv Science

bioRxiv2018

Behavioral effect of chemogenetic inhibition is directly related to receptor transduction levels in rhesus monkeys

We used inhibitory DREADDs (Designer Receptors Exclusively Activated by Designer Drugs) to reversibly disrupt dorsolateral prefrontal cortex (dlPFC) function in male macaque monkeys. Monkeys were tested on a spatial delayed response task to assess working memory function after intramuscular injection of either clozapine-N-oxide (CNO) or vehicle. CNO injections given before DREADD transduction were without effect on behavior. rAAV5/hsyn-hM4Di-mCherry was injected bilaterally into the dlPFC of five male rhesus monkeys, to produce neuronal expression of the inhibitory (Gi-coupled) DREADD receptor. We quantified the percentage of DREADD- transduced cells using stereological analysis of mCherry-immunolabeled cells. We found a greater number of immunolabeled neurons in monkeys that displayed CNO-induced behavioral impairment after DREADD transduction compared to monkeys that showed no behavioral effect after CNO. Even in monkeys that showed reliable effects of CNO on behavior after DREADD transduction, the number of prefrontal neurons transduced with DREADD receptor was on the order of 3% of total prefrontal neurons counted. This level of histological analysis facilitates our understanding of behavioral effects, or lack thereof, after DREADD vector injection in monkeys. It also implies that a functional silencing of a relatively small fraction of dlPFC neurons, albeit in a widely distributed area, is sufficient to disrupt spatial working memory.\n\nSignificance StatementCognitive domains such as working memory and executive function are mediated by the dorsolateral prefrontal cortex (dlPFC). Impairments in these domains are common in neurodegenerative diseases as well as normal aging. The present study sought to measure deficits in a spatial delayed response task following activation of viral-vector transduced inhibitory DREADD (Designer Receptor Exclusively Activated by Designer Drug) receptors in rhesus macaques and compare this to the level of transduction in dlPFC using stereology. We found a significant relationship between the extent of DREADD transduction and the magnitude of behavioral deficit following administration of the DREADD actuator compound clozapine-N- oxide (CNO). These results demonstrate it will be critical to validate transduction to ensure DREADDs remain a powerful tool for neuronal disruption.

neuroscience↗

bioRxiv2018

The rhesus monkey hippocampus critically contributes to scene memory retrieval, but not new learning

Humans can recall a large number of memories years after the events that triggered them. Early studies of humans with amnesia led to the hippocampus being viewed as the critical structure for episodic memory, but human lesions are imprecise, making it difficult to identify the anatomical structures underlying memory impairments. Rodent studies enable great temporal and anatomical precision in hippocampal manipulations, but not investigation of the rich assortment of interleaved memories that occurs in humans. Thus it is not known how lesions restricted to the hippocampus affect the retrieval of multiple sequentially encoded memories. Furthermore, disagreement exists as to whether hippocampal inactivations lead to a temporally graded, or ungraded amnesia, which could be a consequence of different types hippocampal disruption observed in rodent and human studies. In the current study, four rhesus monkeys received bilateral neurotoxic lesions of the hippocampus, and were compared to thirteen unoperated controls on recognition and new learning of visual object-in-place scenes. Monkeys with hippocampal lesions were significantly impaired compared to controls at remembering scenes that were encoded before the lesion. We did not observe any temporal gradient effect of the lesion on memory recognition, with recent and remote memories being equally affected by the lesion. Monkeys with hippocampal lesions showed no deficits in learning and later recognising new scenes. Thus, the hippocampus, like other cortical regions, may be engaged in the acquisition and storage of new memories, but its role can be taken over by spared regions following a lesion. These findings illustrate the utility of experimental paradigms for studying retrograde and anterograde amnesia that make use of the capacity of nonhuman primates to rapidly acquire many distinct visual memories.

neuroscience↗