In this review we show that the cephalopod vertical lobe (VL) provides a good system for assessing the level of evolutionary convergence of the function and organization of neuronal circuitry for mediating learning and memory in animals with complex behavior. The pioneering work of JZ Young described the morphological convergence of the VL with the mammalian hippocampus, cerebellum and the insect mushroom body. Studies in octopus and cuttlefish VL networks suggest evolutionary convergence into a universal organization of connectivity as a divergence-convergence ('fan-out fan-in') network with activity-dependent long-term plasticity mechanisms. Yet, these studies also show that the properties of the neurons, neurotransmitters, neuromodulators and mechanisms of long-term potentiation (LTP) induction and maintenance are highly variable among different species. This suggests that complex networks may have evolved independently multiple times and that even though memory and learning networks share similar organization and cellular processes, there are many molecular ways of constructing them.

The vertical lobe (VL) in the octopus brain plays an essential role in its sophisticated learning and memory. Early anatomical studies suggested that the VL is organized in a “fan‐out fan‐in” connectivity matrix comprising only three morphologically identified neuron types; input axons from the median superior frontal lobe (MSFL) innervating en passant millions of small amacrine interneurons (AMs), which converge sharply onto large VL output neurons (LNs). Recent physiological studies confirmed the feedforward excitatory connectivity; a glutamatergic synapse at the first MSFL‐to‐AM synaptic layer and a cholinergic AM‐to‐LNs synapse. MSFL‐to‐AMs synapses show a robust hippocampal‐like activity‐dependent long‐term potentiation (LTP) of transmitter release. 5‐HT, octopamine, dopamine and nitric oxide modulate short‐ and long‐term VL synaptic plasticity. Here, we present a comprehensive histolabeling study to better characterize the neural elements in the VL. We generally confirmed glutamatergic MSFLs and cholinergic AMs. Intense labeling for NOS activity in the AMs neurites were in‐line with the NO‐dependent presynaptic LTP mechanism at the MSFL‐to‐AM synapse. New discoveries here reveal more heterogeneity of the VL neurons than previously thought. GABAergic AMs suggest a subpopulation of inhibitory interneurons in the first input layer. Clear γ‐amino butyric acid labeling in the cell bodies of LNs supported an inhibitory VL output, yet the LNs co‐expressed FMRFamide‐like neuropeptides, suggesting an additional neuromodulatory role of the VL output. Furthermore, a group of LNs was glutamatergic. A new cluster of cells organized as a “deep nucleus” showed rich catecholaminergic labeling and may play a role in intrinsic neuromodulation. In‐situ hybridization and immunolabeling allowed characterization and localization of a rich array of neuropeptides and neuromodulators, likely involved in reward/punishment signals. This analysis of the fast transmission system, together with the newly found cellular elements, help integrate behavioral, physiological, pharmacological and connectome findings into a more comprehensive understanding of an efficient learning and memory network.

The octopus brain shows a robust hippocampal-like activity-dependent LTP, which is NMDA-independent, yet associative and presynaptically expressed and, as shown here, also independent of protein synthesis. Have the molecular mechanisms for mediating this LTP evolved independently or have they converged? Here we report on a distinctive adaptation of the nitric-oxide (NO) system for mediation of the octopus LTP. Unlike the suggested role of NO in LTP induction in the hippocampus, in octopus, inhibitors of NO-synthase (NOS) did not block LTP induction but either 1) reversibly 'erased' LTP expression, suggesting that a constitutive elevation in NO mediates the presynaptic LTP expression or 2) 'reversed' LTP induction and maintenance because a second LTP could be induced after inhibitor washout. We therefore propose a protein synthesis-independent 'molecular-switch', whereby NO-dependent NOS reactivation maintains NOS in its active state. Thus, while the octopus LTP shows marked evolutionary convergence with LTP in vertebrates, an extreme molecular novelty has evolved to mediate it.

The vertical lobe (VL) in the octopus brain plays an essential role in its sophisticated learning and memory. Early anatomical studies suggested that the VL is organized in a “fan-out fan-in” connectivity matrix comprising only three morphologically identified neuron types; input axons from the superior frontal lobe (SFL) innervating en passant millions of small amacrine interneurons (AMs) which converge sharply onto large VL output neurons (LNs). Recent physiological studies confirmed the feedforward excitatory connectivity: a glutamatergic synapse at the first SFL-to-AM synaptic layer and a cholinergic AM-to-LNs synapse. SFL-to-AMs synapses show a robust hippocampal-like activity-dependent long-term potentiation (LTP) of transmitter release. 5-HT, octopamine, dopamine and nitric oxide modulate short- and long-term VL synaptic plasticity. Here we present a comprehensive histolabeling study to better characterize the neural elements in the VL. We generally confirmed glutamatergic SFLs and cholinergic AMs. Intense labeling for NOS activity in the AMs neurites fitted with the NO-dependent presynaptic LTP mechanism at the SFL-to-AM synapse. New discoveries here reveal more heterogeneity of the VL neurons than previously thought. GABAergic AMs suggest a subpopulation of inhibitory interneurons in the first input layer. Clear GABA labeling in the cell bodies of LNs supported an inhibitory VL output yet the LNs co-expressed FMRFamide-like neuropeptides suggesting an additional neuromodulatory role of the VL output. Furthermore, a group of LNs was glutamatergic. A new cluster of cells organized in a “deep nucleus” showed rich catecholaminergic labeling and may play a role in intrinsic neuromodulation. In situ hybridization and immunolabeling allowed characterization and localization of a rich array of neuropeptides and neuromodulatores, likely involved in reward/punishment signals. This analysis of the fast transmission system, together with the newly found cellular elements helps integrate behavioral, physiological, pharmacological and connectome findings into a more comprehensive understanding of an efficient learning and memory network.