purification-of-the-dendritic-filopodia-rich-fraction

Purification of the Dendritic Filopodia-rich Fraction

Dendritic filopodia are thin and long protrusions based on the actin filament, and they extend and retract as if searching for a target axon. When the ...

RIKEN Center for Brain Science

Dendritic filopodia are thin, long, and highly mobile protrusions functioning as spine precursors. By contrast with a wealth of knowledge on molecular profiles in spines, little is known about structural and functional proteins present in dendritic filopodia. To reveal the molecular constituents of dendritic filopodia, we developed a new method for biochemical preparation of proteins enriched in dendritic filopodia, by taking advantage of specific and strong binding between a dendritic filopodial membrane protein, telencephalin, and its extracellular matrix ligand, vitronectin. When vitronectin-coated magnetic microbeads were added onto cultured hippocampal neurons, phagocytic cup-like membrane protrusions were formed on dendrites through the binding to telencephalin. Magnetically purified membrane protrusion fraction was subjected to comprehensive mass spectrometric analysis and 319 proteins were identified, many of which were confirmed to be localized to dendritic filopodia. Thus, this study provides a useful resource for studying molecular mechanisms underlying dendritic development, synapse formation, and plasticity.

Proteomic Analysis of Dendritic Filopodia-Rich Fraction Isolated by Telencephalin and Vitronectin Interaction.

Mating drive is balanced by a need to safeguard resources for offspring, yet the neural basis for negative regulation of mating remains poorly understood. In rodents, pheromones critically regulate sexual behavior. Here, we observe suppression of adult female sexual behavior in mice by exocrine gland-secreting peptide 22 (ESP22), a lacrimal protein from juvenile mice. ESP22 activates a dedicated vomeronasal receptor, V2Rp4, and V2Rp4 knockout eliminates ESP22 effects on sexual behavior. Genetic tracing of ESP22-responsive neural circuits reveals a critical limbic system connection that inhibits reproductive behavior. Furthermore, V2Rp4 counteracts a highly related vomeronasal receptor, V2Rp5, that detects the male sex pheromone ESP1. Interestingly, V2Rp4 and V2Rp5 are encoded by adjacent genes, yet couple to distinct circuits and mediate opposing effects on female sexual behavior. Collectively, our study reveals molecular and neural mechanisms underlying pheromone-mediated sexual rejection, and more generally, how inputs are routed through olfactory circuits to evoke specific behaviors.

Sexual rejection via a vomeronasal receptor-triggered limbic circuit.

Odorants are recognized by multiple olfactory receptors (ORs) and induce innate behaviors like attraction or aversion via olfactory system in mice. However, a role of an individual OR is unclear. Muscone is recognized by a few ORs including MOR215-1 and MOR214-3, and attracts male mice. Odor preference tests using MOR215-1 knockout mice revealed that MOR215-1 and other OR(s), possibly including MOR214-3, are involved in the attraction. (Z)-5-tetradecen-1-ol (Z5-14:OH) activates ~3 ORs, including Olfr288, and evokes attraction at low levels but aversion at higher levels. Olfr288 knockout mice show no attraction but aversion, suggesting Olfr288 is involved in preference for Z5-14:OH, whereas activation of other low-affinity Z5-14:OH receptors evokes aversion. Each OR appears to send a signal to a neural circuit that possesses distinct valence, leading to a certain behavior. The final output behavior with multiple ORs stimulation is determined by summation (addition or competition) of valences coded by activated ORs.

Article	Year
Proteomic Analysis of Dendritic Filopodia-Rich Fraction Isolated by Telencephalin and Vitronectin Interaction. Frontiers in synaptic neuroscience\| PubMed ID: 30147651	2018
Sexual rejection via a vomeronasal receptor-triggered limbic circuit. Nature communications\| PubMed ID: 30367054	2018
Contribution of individual olfactory receptors to odor-induced attractive or aversive behavior in mice. Nature communications\| PubMed ID: 30643144	2019

Yoshihiro Yoshihara

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