RESUMO
The infiltration of immune cells into tissues underlies the establishment of tissue-resident macrophages and responses to infections and tumors. Yet the mechanisms immune cells utilize to negotiate tissue barriers in living organisms are not well understood, and a role for cortical actin has not been examined. Here, we find that the tissue invasion of Drosophila macrophages, also known as plasmatocytes or hemocytes, utilizes enhanced cortical F-actin levels stimulated by the Drosophila member of the fos proto oncogene transcription factor family (Dfos, Kayak). RNA sequencing analysis and live imaging show that Dfos enhances F-actin levels around the entire macrophage surface by increasing mRNA levels of the membrane spanning molecular scaffold tetraspanin TM4SF, and the actin cross-linking filamin Cheerio, which are themselves required for invasion. Both the filamin and the tetraspanin enhance the cortical activity of Rho1 and the formin Diaphanous and thus the assembly of cortical actin, which is a critical function since expressing a dominant active form of Diaphanous can rescue the Dfos macrophage invasion defect. In vivo imaging shows that Dfos enhances the efficiency of the initial phases of macrophage tissue entry. Genetic evidence argues that this Dfos-induced program in macrophages counteracts the constraint produced by the tension of surrounding tissues and buffers the properties of the macrophage nucleus from affecting tissue entry. We thus identify strengthening the cortical actin cytoskeleton through Dfos as a key process allowing efficient forward movement of an immune cell into surrounding tissues.
Assuntos
Citoesqueleto de Actina/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Macrófagos/fisiologia , Animais , Movimento Celular , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Genes de Insetos , Genes fos , Análise de Sequência de RNA , Tetraspaninas , Fatores de Transcrição/metabolismoRESUMO
Cholecystokinin-expressing interneurons (CCKIs) are hypothesized to shape pyramidal cell-firing patterns and regulate network oscillations and related network state transitions. To directly probe their role in the CA1 region, we silenced their activity using optogenetic and chemogenetic tools in mice. Opto-tagged CCKIs revealed a heterogeneous population, and their optogenetic silencing triggered wide disinhibitory network changes affecting both pyramidal cells and other interneurons. CCKI silencing enhanced pyramidal cell burst firing and altered the temporal coding of place cells: theta phase precession was disrupted, whereas sequence reactivation was enhanced. Chemogenetic CCKI silencing did not alter the acquisition of spatial reference memories on the Morris water maze but enhanced the recall of contextual fear memories and enabled selective recall when similar environments were tested. This work suggests the key involvement of CCKIs in the control of place-cell temporal coding and the formation of contextual memories.
Assuntos
Colecistocinina , Hipocampo , Interneurônios , Optogenética , Células Piramidais , Animais , Masculino , Camundongos , Região CA1 Hipocampal/fisiologia , Região CA1 Hipocampal/citologia , Região CA1 Hipocampal/metabolismo , Colecistocinina/metabolismo , Colecistocinina/genética , Medo/fisiologia , Hipocampo/fisiologia , Interneurônios/fisiologia , Interneurônios/metabolismo , Aprendizagem/fisiologia , Aprendizagem em Labirinto/fisiologia , Memória/fisiologia , Rememoração Mental/fisiologia , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Células Piramidais/fisiologia , Células Piramidais/metabolismo , Ritmo Teta/fisiologiaRESUMO
In vitro work revealed that excitatory synaptic inputs to hippocampal inhibitory interneurons could undergo Hebbian, associative, or non-associative plasticity. Both behavioral and learning-dependent reorganization of these connections has also been demonstrated by measuring spike transmission probabilities in pyramidal cell-interneuron spike cross-correlations that indicate monosynaptic connections. Here we investigated the activity-dependent modification of these connections during exploratory behavior in rats by optogenetically inhibiting pyramidal cell and interneuron subpopulations. Light application and associated firing alteration of pyramidal and interneuron populations led to lasting changes in pyramidal-interneuron connection weights as indicated by spike transmission changes. Spike transmission alterations were predicted by the light-mediated changes in the number of pre- and postsynaptic spike pairing events and by firing rate changes of interneurons but not pyramidal cells. This work demonstrates the presence of activity-dependent associative and non-associative reorganization of pyramidal-interneuron connections triggered by the optogenetic modification of the firing rate and spike synchrony of cells.
Assuntos
Comportamento Exploratório/fisiologia , Hipocampo/fisiologia , Interneurônios/fisiologia , Optogenética , Células Piramidais/fisiologia , Animais , Masculino , Ratos , Ratos Long-EvansRESUMO
Memory consolidation is thought to depend on the reactivation of waking hippocampal firing patterns during sleep. Following goal learning, the reactivation of place cell firing can represent goals and predicts subsequent memory recall. However, it is unclear whether reactivation promotes the recall of the reactivated memories only or triggers wider reorganization. We trained animals to locate goals at fixed locations in two different environments. Following learning, by performing online assembly content decoding, the reactivation of only one environment was disrupted, leading to recall deficit in that environment. The place map of the disrupted environment was destabilized but re-emerged once the goal was relearned. These data demonstrate that sleep reactivation facilitates goal-memory retrieval by strengthening memories that enable the selection of context-specific hippocampal maps. However, sleep reactivation may not be needed for the stabilization of place maps considering that the map of the disrupted environment re-emerged after the retraining of goals.