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• W2794642014 abstract "•A tear protein, ratCRP1, activates the vomeronasal system in female rats and mice•RatCRP1 activates multiple type-2 vomeronasal receptors, including Vmn2r28, in mice•RatCRP1 modulates locomotive activity, body temperature, and heart rate in mice•RatCRP1 acts as an intra-specific signal in rats and an inter-specific signal in mice Rodents use the vomeronasal olfactory system to acquire both inter- and intra-specific information from the external environment and take appropriate actions. For example, urinary proteins from predator species elicit avoidance in mice, while those from male mice attract female mice. In addition to urinary proteins, recent studies have highlighted the importance of lacrimal proteins for intra-specific communications in mice. However, whether the tear fluid of other species also mediates social signals remains unknown. Here, we show that a lacrimal protein in rats (predators of mice), called cystatin-related protein 1 (ratCRP1), activates the vomeronasal system of mice. This protein is specifically produced by adult male rats in a steroid hormone-dependent manner, activates the vomeronasal system of female rats, and enhances stopping behavior. When detected by mice, ratCRP1 activates the medial hypothalamic defensive circuit, resulting in decreased locomotion coupled with lowered body temperature and heart rate. Notably, ratCRP1 is recognized by multiple murine type 2 vomeronasal receptors, including Vmn2r28. CRISPR/Cas9-mediated deletion of vmn2r28 impaired both ratCRP1-induced neural activation of the hypothalamic center and decrease of locomotor activity in mice. Taken together, these data reveal the neural and molecular basis by which a tear fluid compound in rats affects the behavior of mice. Furthermore, our study reveals a case in which a single compound that mediates an intra-specific signal in a predator species also functions as an inter-specific signal in the prey species. Rodents use the vomeronasal olfactory system to acquire both inter- and intra-specific information from the external environment and take appropriate actions. For example, urinary proteins from predator species elicit avoidance in mice, while those from male mice attract female mice. In addition to urinary proteins, recent studies have highlighted the importance of lacrimal proteins for intra-specific communications in mice. However, whether the tear fluid of other species also mediates social signals remains unknown. Here, we show that a lacrimal protein in rats (predators of mice), called cystatin-related protein 1 (ratCRP1), activates the vomeronasal system of mice. This protein is specifically produced by adult male rats in a steroid hormone-dependent manner, activates the vomeronasal system of female rats, and enhances stopping behavior. When detected by mice, ratCRP1 activates the medial hypothalamic defensive circuit, resulting in decreased locomotion coupled with lowered body temperature and heart rate. Notably, ratCRP1 is recognized by multiple murine type 2 vomeronasal receptors, including Vmn2r28. CRISPR/Cas9-mediated deletion of vmn2r28 impaired both ratCRP1-induced neural activation of the hypothalamic center and decrease of locomotor activity in mice. Taken together, these data reveal the neural and molecular basis by which a tear fluid compound in rats affects the behavior of mice. Furthermore, our study reveals a case in which a single compound that mediates an intra-specific signal in a predator species also functions as an inter-specific signal in the prey species. Many animals depend primarily on olfaction, of the five major senses, to obtain information about their surroundings. These olfactory cues guide them to nesting areas and food, as well as warn them if predators are present. The presence of volatile odorants emitted from predators means that predators are nearby, cueing the animal to prepare for an immediate response, such as escape or battle. In contrast, when the animal sniffs and detects non-volatile or long-lasting volatile cues derived from predators, these cues suggest that the animal has entered the predator’s territory or that the predator may have been present recently. Unlike visual and auditory information that only convey current information occurring within a relatively short distance, chemical signals represent both temporal and spatial information that animals can effectively use to take appropriate action [1Wyatt T.D. Pheromones and Animal Behaviour.Second Edition. Cambridge University Press, Cambridge, UK2014Google Scholar]. In rodents, chemical cues are detected by two distinct olfactory sensory systems: the main olfactory system and the vomeronasal system. Whereas the main olfactory system detects volatile odor cues, the vomeronasal system detects mainly non-volatile cues. The olfactory and vomeronasal systems send information first to the main and accessory olfactory bulbs, respectively, and then to the amygdala and hypothalamus in the brain, thus evoking various innate immediate behaviors (called “releasing effects”) and endocrine or emotional changes (called “priming effects”) [2Chamero P. Leinders-Zufall T. Zufall F. From genes to social communication: molecular sensing by the vomeronasal organ.Trends Neurosci. 2012; 35: 597-606Abstract Full Text Full Text PDF PubMed Scopus (107) Google Scholar, 3Liberles S.D. Mammalian pheromones.Annu. Rev. Physiol. 2014; 76: 151-175Crossref PubMed Scopus (178) Google Scholar, 4Stowers L. Kuo T.H. Mammalian pheromones: emerging properties and mechanisms of detection.Curr. Opin. Neurobiol. 2015; 34: 103-109Crossref PubMed Scopus (66) Google Scholar]. Although volatile odor cues involved in chemical communication and their underlying mechanisms for specific behavioral output have been widely studied, the molecular bases for releasing or priming effects by non-volatile cues, such as peptides and proteins, have not. Urine appears to be a major source of vomeronasal signals. For example, the major urinary protein (MUP) family is involved in individual recognition within the same species in rodents. Some MUPs promote aggression in male mice [5Chamero P. Marton T.F. Logan D.W. Flanagan K. Cruz J.R. Saghatelian A. Cravatt B.F. Stowers L. Identification of protein pheromones that promote aggressive behaviour.Nature. 2007; 450: 899-902Crossref PubMed Scopus (401) Google Scholar], and one male-specific MUP attracts female mice [6Roberts S.A. Simpson D.M. Armstrong S.D. Davidson A.J. Robertson D.H. McLean L. Beynon R.J. Hurst J.L. Darcin: a male pheromone that stimulates female memory and sexual attraction to an individual male’s odour.BMC Biol. 2010; 75Google Scholar, 7Roberts S.A. Davidson A.J. McLean L. Beynon R.J. Hurst J.L. Pheromonal induction of spatial learning in mice.Science. 2012; 338: 1462-1465Crossref PubMed Scopus (119) Google Scholar, 8Kaur A.W. Ackels T. Kuo T.H. Cichy A. Dey S. Hays C. Kateri M. Logan D.W. Marton T.F. Spehr M. Stowers L. Murine pheromone proteins constitute a context-dependent combinatorial code governing multiple social behaviors.Cell. 2014; 157: 676-688Abstract Full Text Full Text PDF PubMed Scopus (129) Google Scholar]. Therefore, MUPs are utilized for intra-specific communication and act as protein pheromones. MUPs can also function as inter-specific signals. Rat or cat MUPs activate mouse vomeronasal sensory neurons (VSNs) and evoke avoidance behavior [9Isogai Y. Si S. Pont-Lezica L. Tan T. Kapoor V. Murthy V.N. Dulac C. Molecular organization of vomeronasal chemoreception.Nature. 2011; 478: 241-245Crossref PubMed Scopus (221) Google Scholar, 10Papes F. Logan D.W. Stowers L. The vomeronasal organ mediates interspecies defensive behaviors through detection of protein pheromone homologs.Cell. 2010; 141: 692-703Abstract Full Text Full Text PDF PubMed Scopus (243) Google Scholar, 11Pérez-Gómez A. Bleymehl K. Stein B. Pyrski M. Birnbaumer L. Munger S.D. Leinders-Zufall T. Zufall F. Chamero P. Innate Predator Odor Aversion Driven by Parallel Olfactory Subsystems that Converge in the Ventromedial Hypothalamus.Curr. Biol. 2015; 25: 1340-1346Abstract Full Text Full Text PDF PubMed Scopus (105) Google Scholar]. Thus, urine contains vomeronasal ligands used for intra- or inter-specific communication. Recently, it has been revealed that secretions in facial areas, such as tear fluid and saliva, are also sources of signals. For instance, a peptide pheromone, exocrine gland-secreting peptide 1 (ESP1), is secreted into male mouse tear fluid. ESP1 is detected by a single member of the vomeronasal receptor type 2 family, V2Rp5 (also known as Vmn2r116), in mice, leading to enhanced sexual receptive behavior in female mice and aggression in male mice [12Haga S. Hattori T. Sato T. Sato K. Matsuda S. Kobayakawa R. Sakano H. Yoshihara Y. Kikusui T. Touhara K. The male mouse pheromone ESP1 enhances female sexual receptive behaviour through a specific vomeronasal receptor.Nature. 2010; 466: 118-122Crossref PubMed Scopus (278) Google Scholar, 13Hattori T. Osakada T. Matsumoto A. Matsuo N. Haga-Yamanaka S. Nishida T. Mori Y. Mogi K. Touhara K. Kikusui T. Self-Exposure to the Male Pheromone ESP1 Enhances Male Aggressiveness in Mice.Curr. Biol. 2016; 26: 1229-1234Abstract Full Text Full Text PDF PubMed Scopus (29) Google Scholar, 14Kimoto H. Haga S. Sato K. Touhara K. Sex-specific peptides from exocrine glands stimulate mouse vomeronasal sensory neurons.Nature. 2005; 437: 898-901Crossref PubMed Scopus (290) Google Scholar, 15Kimoto H. Sato K. Nodari F. Haga S. Holy T.E. Touhara K. Sex- and strain-specific expression and vomeronasal activity of mouse ESP family peptides.Curr. Biol. 2007; 17: 1879-1884Abstract Full Text Full Text PDF PubMed Scopus (116) Google Scholar]. In humans, tear fluid is also proposed to be a source of chemical cues that affect emotion; exposure of negative emotional tear fluid from women results in reduction of sexual arousal in receiver men [16Gelstein S. Yeshurun Y. Rozenkrantz L. Shushan S. Frumin I. Roth Y. Sobel N. Human tears contain a chemosignal.Science. 2011; 331: 226-230Crossref PubMed Scopus (149) Google Scholar]. Tears appear to convey intra-specific information in various animals. But the functions of tear fluid relating to inter-specific communication have never been investigated. In this study, we hypothesized that tear fluid could also be a source of inter-specific signals. This possibility was examined by focusing on a well-accepted predator-prey model: the rat and mouse. Using an immunohistochemistry-assisted purification strategy in combination with behavioral and physiological experiments, we discovered a protein with previously unknown function in the tear fluid of male rats. This discovery led us to study the functions of this protein in both rats and mice, as well as the neural and molecular bases by which the mouse vomeronasal system detects this lacrimal protein of predators. To examine whether rat tears contain substances that activate VSNs in mice, we immunolabeled the vomeronasal epithelium from a rat tear-stimulated mouse with anti-phosphorylated ribosomal protein S6 (pS6) as a marker for neural activation [17Abe T. Touhara K. Structure and function of a peptide pheromone family that stimulate the vomeronasal sensory system in mice.Biochem. Soc. Trans. 2014; 42: 873-877Crossref PubMed Scopus (16) Google Scholar, 18Knight Z.A. Tan K. Birsoy K. Schmidt S. Garrison J.L. Wysocki R.W. Emiliano A. Ekstrand M.I. Friedman J.M. Molecular profiling of activated neurons by phosphorylated ribosome capture.Cell. 2012; 151: 1126-1137Abstract Full Text Full Text PDF PubMed Scopus (192) Google Scholar]. In both male and female BALB/c mice stimulated with male rat tear fluid (containing 1000 μg of protein equivalent to 33 μl of tear fluid), pS6-positive VSNs were observed in the basal zone of the vomeronasal epithelium (male: 72 ± 6, female: 80 ± 3; mean per section ± SE) (Figures 1A and 1B). In contrast, a smaller number of pS6-positive VSNs were observed upon stimulation with female rat tear fluid (male: 10 ± 2, female: 9 ± 3). We next examined whether the male rat tear signal was relayed to the accessory olfactory bulb (AOB) to which VSN axons directly project by using c-Fos as a marker [12Haga S. Hattori T. Sato T. Sato K. Matsuda S. Kobayakawa R. Sakano H. Yoshihara Y. Kikusui T. Touhara K. The male mouse pheromone ESP1 enhances female sexual receptive behaviour through a specific vomeronasal receptor.Nature. 2010; 466: 118-122Crossref PubMed Scopus (278) Google Scholar]. c-Fos-positive cells were observed in the AOB mitral/tufted cell layer of mice stimulated with male rat tear fluid (204 ± 42 per AOB) (Figures 1C and 1D). The c-Fos-positive cells were located in the posterior part of the AOB, suggesting that rat tear fluid activates VSNs expressing V2R-type receptors whose ligands are peptides or proteins [19Touhara K. Vosshall L.B. Sensing odorants and pheromones with chemosensory receptors.Annu. Rev. Physiol. 2009; 71: 307-332Crossref PubMed Scopus (397) Google Scholar]. Dose-dependent increases in the number of c-Fos-positive cells were observed with a threshold of approximately 1–10 μg of tear proteins (Figure 1E). Together, these data demonstrate that male rat tear fluid contains peptide or protein cues that activate the mouse vomeronasal neural system. To identify the active compound(s), we fractionated male rat tear fluid using high-performance liquid chromatography (HPLC) with a C4 reverse-phase column, and each fraction was tested for AOB c-Fos-inducing activity. The activity was eluted at 32.5–35.0 min (36.2%–37.2% acetonitrile; fraction 4 in Figure 2A). We next applied fraction 4 to a C30 column. c-Fos-induced activity was found in the eluents at 32.0–40.0 min (36.0%–39.2% acetonitrile) (fraction e in Figure 2A). The amino-terminal peptide sequence of fraction e was determined. A genome search of this sequence identified the active component as rat cystatin-related protein 1 (ratCRP1), a 21-kDa protein with unknown function. RatCRP1 belongs to the cystatin gene superfamily, of which some members encode for proteinase inhibitors (Figure S1A) [20Devos A. De Clercq N. Vercaeren I. Heyns W. Rombauts W. Peeters B. Structure of rat genes encoding androgen-regulated cystatin-related proteins (CRPs): a new member of the cystatin superfamily.Gene. 1993; 125: 159-167Crossref PubMed Scopus (28) Google Scholar]. We next verified that recombinant ratCRP1 could stimulate the vomeronasal system of mice (Figure S1B). We observed many pS6-positive cells in the vomeronasal epithelium (Figures 2B and 2C). The number of pS6-positive cells (77 ± 9 per slice) was approximately the same as that observed upon stimulation with male rat tear fluid (Figures 1B and 2C). No pS6-positive cells were found in samples from mice stimulated with a recombinant ratCRP2 that was most closely related to ratCRP1 (92% base sequence similarity) (Figures 2C and S1A). Recombinant ratCRP1 also induced c-Fos expression in AOB mitral/tufted cells in a dose-dependent manner (Figure 2D). c-Fos induction was not observed in mice lacking transient receptor potential C2 (TRPC2), the primary signal transduction channel of VSNs (Figures 2E and 2F) [21Leypold B.G. Yu C.R. Leinders-Zufall T. Kim M.M. Zufall F. Axel R. Altered sexual and social behaviors in trp2 mutant mice.Proc. Natl. Acad. Sci. USA. 2002; 99: 6376-6381Crossref PubMed Scopus (453) Google Scholar, 22Stowers L. Holy T.E. Meister M. Dulac C. Koentges G. Loss of sex discrimination and male-male aggression in mice deficient for TRP2.Science. 2002; 295: 1493-1500Crossref PubMed Scopus (666) Google Scholar]. Together, these results suggest that ratCRP1 is the major component of male rat tear fluid that activates mouse VSNs. We performed reverse-transcriptase polymerase chain reaction (RT-PCR) analysis to examine the expression profile of the ratCRP1 gene. Among exocrine glands that secrete tear fluid (i.e., the extraorbital lacrimal gland [ELG] and the Hardarian gland [HG]) and saliva (i.e., the parotid gland [PG], the sublingual gland [SLG], and the submaxillary gland [SMG]), ratCRP1 was expressed in ELG, PG, and SLG (Figure 3A). RatCRP1 was also expressed in testis and prostate, consistent with previous studies [23Winderickx J. Hemschoote K. De Clercq N. Van Dijck P. Peeters B. Rombauts W. Verhoeven G. Heyns W. Tissue-specific expression and androgen regulation of different genes encoding rat prostatic 22-kilodalton glycoproteins homologous to human and rat cystatin.Mol. Endocrinol. 1990; 4: 657-667Crossref PubMed Scopus (52) Google Scholar]. RatCRP1 expression in ELG begins 3 weeks after birth and continues to be expressed thereafter only in male rats (Figure 3B). Castrated 3-week-old male rats did not express ratCRP1, whereas testosterone treatment induced ratCRP1 expression in female rats (Figure 3C). The expression profile for ratCRP2 was similar to that of ratCRP1, whereas other cystatin family genes exhibited dissimilar expression profiles (Figure S2). These results indicate that ratCRP1 is expressed in exocrine glands in a testosterone-dependent manner and thus secreted in the tears of male rats, specifically. To estimate the amount of ratCRP1 secreted in male rat tear fluid, we produced an anti-ratCRP1 antibody and performed western blot analysis. The ratCRP1 protein was detected in ELG and male tear fluid but not in other exocrine glands or female tear fluid (Figures 3D and 3E). RatCRP1 was expressed in male tear fluid of various rat strains (Figure 3F). The amount of ratCRP1 in male rat tear fluid was estimated to be about 1/10 of total tear fluid protein (Figure S3). This value is fairly consistent with the threshold difference in mitral/tufted cell activation between tear fluid proteins and ratCRP1 (Figures 1E and 2D). Considering that the protein concentration of male rat tear fluid is approximately 30 ± 1 μg/μL, 1 μL of tear fluid, which can be easily collected with a single pipetting from a rat eye, contains 3 μg of ratCRP1. This amount is enough to stimulate three mice. Therefore, rats secrete a fairly large amount of ratCRP1, which is transferred to their paws, legs, and fur during grooming and is eventually deposited within the rat’s territory area, where mice may enter and sniff. Why do only male rats substantially secrete ratCRP1 in a testosterone-dependent manner? We hypothesized that ratCRP1 may act as a pheromone that influences sexual behavior or inter-male aggression via activation of the vomeronasal system. We thus assessed whether ratCRP1 evokes c-Fos induction in the male and female rat AOB. In female rats stimulated with recombinant ratCRP1, we observed many c-Fos-positive cells in the AOB mitral/tufted cell layer (8 ± 3 per section) (Figures 4A and 4B). In contrast, no significant c-Fos induction was observed in the AOB of male rats (1 ± 1 per section) (Figures 4A and 4B). We also examined higher brain regions beyond the AOB and found that stimulation with ratCRP1 resulted in induction of c-Fos expression in the ventromedial hypothalamus (VMH) in female rats (22 ± 2 per section) (Figures 4C and 4D). These results suggest that ratCRP1 from male rats conveys a signal to higher vomeronasal centers of female rats. We next examined the behavioral effects of ratCRP1 in female rats. Female rats sniffed intact male rat bedding longer than control bedding, yet addition of ratCRP1 to new bedding or the bedding of castrated male rats did not evoke interest in female rats (Figure 4E). Nevertheless, we noticed that 10 min after stimulation with ratCRP1 cotton, female rats often stopped moving for a while. We defined stopping as a halting posture without sniffing. The total duration of stopping was significantly increased by ratCRP1, while the duration of cotton investigation, grooming, or standing was unchanged (Figures 4F–4J). In typical sexual behavior, the estrus female rat comes to an abrupt halt—similar to ratCRP1-stimulated stopping—at the end of a locomotor sequence that includes “hopping and darting” and usually assumes a crouching posture [24Pfaff D.W. Estrogens and brain function. Springer-Verlag, 1980Crossref Google Scholar]. This facilitates mounting of the male rat, followed by the lordosis response of the female rat. We thus tested the effects of ratCRP1 in sexual behaviors of female rats. Prior exposure to ratCRP1, however, did not increase lordosis in female rats (Figure S4). In summary, our data revealed that ratCRP1 produced by male rats activated the vomeronasal system of female rats and increased their stopping behavior. How does the mouse vomeronasal system detect ratCRP1? To answer this question, we aimed to identify the vomeronasal receptor(s) of ratCRP1 in mice. Vomeronasal receptors comprise two multigene families, V1R and V2R, which are encoded by 187 and 121 intact genes, respectively, in mice [25Young J.M. Trask B.J. V2R gene families degenerated in primates, dog and cow, but expanded in opossum.Trends Genet. 2007; 23: 212-215Abstract Full Text Full Text PDF PubMed Scopus (133) Google Scholar, 26Nei M. Niimura Y. Nozawa M. The evolution of animal chemosensory receptor gene repertoires: roles of chance and necessity.Nat. Rev. Genet. 2008; 9: 951-963Crossref PubMed Scopus (395) Google Scholar]. Proteinaceous signals are detected by V2Rs. The vomeronasal organs (VNOs) of mice stimulated with ratCRP1 were analyzed by pS6 immunohistochemistry coupled with in situ hybridization (ISH) with cRNA probes that recognize various V2R clades. Of the pS6-positive neurons activated by ratCRP1, 96% co-labeled with V2Ra probe signals (a1+a2) but not with signals from other probes (Figures 5A, 5B, S5, and S6A). Approximately 47% of V2Ra-expressing neurons were pS6-positive (Figures 5A, 5B, and S6A). Using more specific V2Ra probes, we found that pS6-positive neurons labeled with both a1 and a2 probe signals (Figure 5C). The a1 probe can recognize both vmn2r28 and vmn2r52 (Figure S5); however, it has been suggested that vmn2r28 is more abundantly expressed in the mouse VNO [27Ibarra-Soria X. Levitin M.O. Saraiva L.R. Logan D.W. The olfactory transcriptomes of mice.PLoS Genet. 2014; 10: e1004593Crossref PubMed Scopus (96) Google Scholar]. We thus generated a Vmn2r28 antibody for use in a double-label immunohistochemistry assay coupled with the pS6 antibody. Over 81% of cells labeled by the Vmn2r28 antibody were pS6-positive after exposure to ratCRP1 (Figures 5D, 5E, and S6B). Vmn2r28-positive cells were pS6-positive when mice were stimulated with rat bedding or tears but not with rat saliva or urine (Figures 5E and S6B). These results indicate that Vmn2r28 is a ratCRP1 receptor in mice. We then generated vmn2r28-deficient mice using the CRISPR/Cas9 system (Figure S7) [28Gaj T. Gersbach C.A. Barbas 3rd, C.F. ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering.Trends Biotechnol. 2013; 31: 397-405Abstract Full Text Full Text PDF PubMed Scopus (2516) Google Scholar]. Double-label immunohistochemistry using pS6 and Vmn2r28 antibodies showed the specific loss of Vmn2r28 expression and a decrease in the number of pS6-positive cells in vmn2r28−/− homozygous mice (Figures 5F and 5G). Notably, many pS6-positive cells still remained in the VNO of ratCRP1-activated vmn2r28-deficient mice (Figures 5F and 5G), clearly demonstrating that there are likely to be additional ratCRP1 receptors in mice. In mice that detect ratCRP1 in the VNO, how is this information represented in the brain? To answer this question, we examined neuronal activation patterns in brain regions that received inputs from the AOB. Stimulation with ratCRP1 resulted in induction of c-Fos expression in the medial amygdala ventral part (MeAv) and VMH dorsal part (VMHd) in both male and female mice (Figures 6A–6D). In the VMHd, c-Fos-positive cells overlapped with neurons expressing steroidogenic factor 1 (SF1), an orphan nuclear receptor gene that is known to be a genetic marker of VMHd neurons (85 ± 5% in male and 92 ± 1% in female mice) (Figures 6B and 6D). We also investigated the c-Fos expression pattern in the VMHd of vmn2r28-deficient mice. Whereas an increase in SF1-positive, c-Fos-positive cells was observed in the VMHd of ratCRP1-stimulated vmn2r28-heterozygous male mice, no such increase was observed in vmn2r28-deficient mice (Figures 6E and 6F). These data suggest that ratCRP1-mediated activation of SF1-expressing neurons requires functional Vmn2r28 receptors. Since SF1-expressing neurons have been indicated to be involved in defensive behaviors [29Kunwar P.S. Zelikowsky M. Remedios R. Cai H. Yilmaz M. Meister M. Anderson D.J. Ventromedial hypothalamic neurons control a defensive emotion state.eLife. 2015; 4https://doi.org/10.7554/eLife.06633Crossref PubMed Scopus (122) Google Scholar, 30Silva B.A. Mattucci C. Krzywkowski P. Murana E. Illarionova A. Grinevich V. Canteras N.S. Ragozzino D. Gross C.T. Independent hypothalamic circuits for social and predator fear.Nat. Neurosci. 2013; 16: 1731-1733Crossref PubMed Scopus (137) Google Scholar, 31Wang L. Chen I.Z. Lin D. Collateral pathways from the ventromedial hypothalamus mediate defensive behaviors.Neuron. 2015; 85: 1344-1358Abstract Full Text Full Text PDF PubMed Scopus (159) Google Scholar], it is reasonable to speculate that ratCRP1 is a predator signal for mice. Since SF1-expressing neurons have been shown to be activated by various predator cues, we analyzed the neural representation of ratCRP1 using cellar compartment analysis of temporal activity by fluorescence ISH (catFISH), with snake skin acting as a reference [32Ishii K.K. Osakada T. Mori H. Miyasaka N. Yoshihara Y. Miyamichi K. Touhara K. A Labeled-Line Neural Circuit for Pheromone-Mediated Sexual Behaviors in Mice.Neuron. 2017; 95: 123-137Abstract Full Text Full Text PDF PubMed Scopus (88) Google Scholar]. Unidentified compounds in snake skin activate mouse V2Rs and induce risk assessment behaviors in a manner that is dependent on a functional VNO [9Isogai Y. Si S. Pont-Lezica L. Tan T. Kapoor V. Murthy V.N. Dulac C. Molecular organization of vomeronasal chemoreception.Nature. 2011; 478: 241-245Crossref PubMed Scopus (221) Google Scholar, 10Papes F. Logan D.W. Stowers L. The vomeronasal organ mediates interspecies defensive behaviors through detection of protein pheromone homologs.Cell. 2010; 141: 692-703Abstract Full Text Full Text PDF PubMed Scopus (243) Google Scholar]. We found that less than 20% of ratCRP1-activated neurons are coincident with snake skin-activated neurons (Figures 6G and 6H), suggesting that a unique subpopulation of SF1-expressing neurons is activated by ratCRP1. What are the behavioral or physiological responses of mice to ratCRP1? Some predator signals are known to evoke immediate responses, such as avoidance or risk assessment behaviors [10Papes F. Logan D.W. Stowers L. The vomeronasal organ mediates interspecies defensive behaviors through detection of protein pheromone homologs.Cell. 2010; 141: 692-703Abstract Full Text Full Text PDF PubMed Scopus (243) Google Scholar, 33Apfelbach R. Blanchard C.D. Blanchard R.J. Hayes R.A. McGregor I.S. The effects of predator odors in mammalian prey species: a review of field and laboratory studies.Neurosci. Biobehav. Rev. 2005; 29: 1123-1144Crossref PubMed Scopus (601) Google Scholar, 34Katz L.B. Dill L.M. The scent of death: Chemosensory assessment of predation risk by animals.Ecoscience. 1998; 5: 361-394Crossref Scopus (1153) Google Scholar]. Thus, we first performed avoidance and risk assessment tests [10Papes F. Logan D.W. Stowers L. The vomeronasal organ mediates interspecies defensive behaviors through detection of protein pheromone homologs.Cell. 2010; 141: 692-703Abstract Full Text Full Text PDF PubMed Scopus (243) Google Scholar, 35Kobayakawa K. Kobayakawa R. Matsumoto H. Oka Y. Imai T. Ikawa M. Okabe M. Ikeda T. Itohara S. Kikusui T. et al.Innate versus learned odour processing in the mouse olfactory bulb.Nature. 2007; 450: 503-508Crossref PubMed Scopus (472) Google Scholar] using samples of cat fur, snake skin, rat urine, ratCRP1, or a ratCRP1/rat urine mixture. Each sample was deposited in one area (zone A) of a plastic box with three compartments, and male mice were placed in another area of the box (zone B). We measured the time each mouse stayed in each zone and tallied the risk assessment behaviors observed in a 5-min period immediately after encountering the stimuli. Mice showed significant avoidance behavior toward snake skin and a trend toward avoidance of cat fur, rat urine, and the ratCRP1/rat urine mixture (Figure 7A). No avoidance behavior was observed toward ratCRP1 (Figure 7A). Snake skin and cat fur induced a transient risk assessment behavior in mice, whereas ratCRP1 did not (Figure 7B). The result is consistent with the presence of neural populations dedicated to ratCRP1 and snake skin processing (Figures 6G and 6H). We also performed the aggressive biting test [36Kuchiiwa S. Kuchiiwa T. A novel semi-automated apparatus for measurement of aggressive biting behavior in mice.J. Neurosci. Methods. 2014; 228: 27-34Crossref PubMed Scopus (12) Google Scholar]. Although male BALB/c mice stimulated with soiled rat bedding tended to exhibit aggressive biting behavior, ratCRP1 did not induce aggressive biting (Figure 7C). These data suggest that ratCRP1 does not induce defensive behaviors, such as avoidance or aggressive biting, that are evoked by other predator cues. We next hypothesized that ratCRP1 does not evoke immediate behaviors, but rather evokes relatively long-lasting, priming effects in mice. We first observed the behaviors of mice when they were touched or bit or sniffed cotton to which ratCRP1 was applied. We found that the time that mice spent investigating ratCRP1-soaked cotton was significantly less than that spent investigating untreated cotton (Figure 7D), which is in contrast to our observat" @default.
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• W2794642014 date "2018-04-01" @default.
• W2794642014 modified "2023-10-13" @default.
• W2794642014 title "Identification of an Intra- and Inter-specific Tear Protein Signal in Rodents" @default.
• W2794642014 cites W126353083 @default.
• W2794642014 cites W1522531331 @default.
• W2794642014 cites W1969464285 @default.
• W2794642014 cites W1976391205 @default.
• W2794642014 cites W1984373254 @default.
• W2794642014 cites W1993469866 @default.
• W2794642014 cites W1999633796 @default.
• W2794642014 cites W2003769343 @default.
• W2794642014 cites W2007507157 @default.
• W2794642014 cites W2009481857 @default.
• W2794642014 cites W2012847399 @default.
• W2794642014 cites W2018710614 @default.
• W2794642014 cites W2039275589 @default.
• W2794642014 cites W2041519359 @default.
• W2794642014 cites W2042656725 @default.
• W2794642014 cites W2047223461 @default.
• W2794642014 cites W2057329083 @default.
• W2794642014 cites W2069493503 @default.
• W2794642014 cites W2071411229 @default.
• W2794642014 cites W2072008258 @default.
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• W2794642014 cites W2171295428 @default.
• W2794642014 cites W2304421282 @default.
• W2794642014 cites W2341078042 @default.
• W2794642014 cites W2646486909 @default.
• W2794642014 cites W2729451008 @default.
• W2794642014 cites W4211192430 @default.
• W2794642014 cites W4233312985 @default.
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