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Affiliative Social Interactions Activate Vasopressin-Responsive Neurons in the Mouse Dorsal Raphe

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Affiliative Social Interactions Activate Vasopressin-Responsive Neurons in the Mouse Dorsal Raphe Rowan University Rowan Digital Works Stratford Campus Research Day 25th Annual Research Day May 6th, 12:00 AM Affiliative Social Interactions Activate Vasopressin-Responsive Neurons in the Mouse Dorsal Raphe Tirth Patel Rowan University Hanna O. Caiola Rowan University Olivia Mallari Rowan University Benjamin D. Rood Rowan University Follow this and additional works at: https://rdw.rowan.edu/stratford_research_day Part of the Animals Commons, Medical Cell Biology Commons, Neurosciences Commons, Other Social and Behavioral Sciences Commons, and the Physiological Processes Commons Let us know how access to this document benefits ouy - share your thoughts on our feedback form. Patel, Tirth; Caiola, Hanna O.; Mallari, Olivia; and Rood, Benjamin D., "Affiliative Social Interactions Activate Vasopressin-Responsive Neurons in the Mouse Dorsal Raphe" (2021). Stratford Campus Research Day. 6. https://rdw.rowan.edu/stratford_research_day/2021/may6/6 This Poster is brought to you for free and open access by the Conferences, Events, and Symposia at Rowan Digital Works. It has been accepted for inclusion in Stratford Campus Research Day by an authorized administrator of Rowan Digital Works. Affilia�ve Social Interac�ons Ac�vate Vasopressin-Responsive Neurons in the Mouse Dorsal Raphe Graduate School of Tirth Patel, Hanna O. Caiola , Olivia Mallari, B. D. Rood Biomedical Sciences Department of Cell Biology and Neuroscience, Rowan University, GSBS, Stratford, NJ Introduction In situ hybridization data suggests that Avpr1a-GFP mouse: GFP expression mirrors Avpr1a-GFP neurons are highly excitable Social behavior is inextricably linked to human health shaping both our susceptibility Avpr1a activation in the DR may be stimulus AVP innervation zones and GFP+ neurons re- compared to multiple subtypes of neighboring and resilience to disease and stress. Positive interactions as simple as maternal contact or friendships among children and adults can protect against emotional dis- specific spond to vasopressin Coronal section through thal- serotonin neurons tress and improve treatment outcomes, whereas negative interactions such as amus illustrates mirroring of 45 abuse, social isolation, or bullying can increase aggression and precipitate mood male male Rood & De Vries (2011) Aq Avpr1a-GFP female vasopresisn innervation and 40 disorders. Discovering the structure and function of neural circuits underlying social 250 PV Avpr1a-GFP neurons Drd2/5-HT behavior is critical to understanding the link between social interaction and health. 35 Non-Drd2/5-HT 200 The neuropeptide vasopressin has been implicated in the regulation of multiple MD MD 30 social interactions including social memory, aggression, mating, pair-bonding, and parental care. Vasopressin producing neurons in the bed nucleus of the stria termi- 150 Vasopressin induces inward 25 (excitatory) currents in patched nalis (BNST) and medial amygdala (MeA), in particular, are predicted to be involved 20 in social behavior. While the innervation targets of BNST and MeA vasopressin neu- 100 Avpr1a-GFP neurons rons and patterns of vasopressin receptor binding have been well-documented in Total Fos+ Cells 20 pA 20 s 15 multiple species, the identity and functional characteristics of neurons targeted by 50 Re 200 nM Vasopressin vasopressin innervation are less well understood. 500 ms current pulse 10 opposite sex homecage 0 0 pA Number of Spikes during same handling homecage opposite Fos sm 5 sex sex Vasopressin Social Behavior Network Serotonin (TPH+) sm Lateral Both female and male subject mice increase fos expression in the Avpr1A 20 40 60 80 100 120 140 160 180 Habenula Dorsal Raphe when exposed to a female stimulus animal. f f Injected Current (pA) Wildtype Avpr1a-GFP Dorsal Lateral Raphe Vasopressin GFP Septum TPH-Avpr1a-Fos+ Bed Nucleus Male Subject of the Stria Mediodorsal 140 TPH-Fos+ Terminalis Thalamus Cells expressing GFP in the 120 Avpr1a-Fos+ Avpr1a-GFP mouse are found primarily Median Fos+ Parastrial Nucleus Raphe 100 in location where vasopressin innerva- Medial Preoptic Area SON Ventral Ventral Pallidum tion have been previously described. Tegmental 80 Lateral Area Hypothalamus Medial Drd2/5HT and Amygdala 60 GFP-expressing neurons Non-Drd2/5-HT Total Fos+ Cells 40 in mid DR 20 Vasopressin 60 Mating 0 innervation in the A Aggression same sex handling homecage opposite sex Serotonin neuron subtypes Handling dorsal raphe 50 Homecage Female Subject TPH-Avpr1a-Fos+ 140 TPH-Fos+ DR Fos expression in BNST of male Avpr1a-Fos+ 40 120 mouse exposed to female mouse Fos+ B 100 -60 mV (Ho et al., 2010) B B 80 The immediate early gene, c-Fos, 30 used as a proxy for neural activa- AB AB 60 C Fos expression in dorsal raphe of male Number of Fos+ Cells A tion is expressed in response to a 20 mouse exposed to female mouse Total Fos+ Cells 40 female, but not an unfamiliar male 20 in both BNST vasopressin neurons 10 0 Avpr1a and in the dorsal raphe, a target of same sex handling homecage opposite sex Drd2/5-HT the BNST vasopressin system. Traces were generated by inject- Express the dopamine Non-Drd2/5-HT Overall increase in Fos expression induced by exposure to a female stimulus Coronal brain slice through rostral dorsal raphe Vasopressin and Tryptophan Hydroxylase ing currents from -80 pA to 180 pA 20 mV Rostral Caudal D2 receptor animal occurs largely in Avpr1a and non-Avpr1a, non-serotonergic neurons. in 20 pA steps 50 ms Ano1 Cacna1a Who are the vasopressin responsive neurons of Cacna1b Action potentials differ between vasopressin-re- Cacna1c Future directions: Use transcriptome Cacna1d Cacna1e Results & Conclusions the dorsal raphe? sponsive neurons and serotonergic neighbors Cacna1f analysis to identify genes that under- Cacna1g Vasopressin neurons in the extended amygdala are activated by mating stimuli. A Cacna1h Cacna1i subset of neurons in the dorsal raphe, a target region of extended amygdala vaso- Cerebral 70 Cacna1s lie Avpr1a-neuron excitability and Kcna1 Aqueduct Kcna10 pressin neurons, is also activated by mating as measured by increases in c-fos. 60 Kcna2 Avpr1a Kcna3 neurochemical phenotype. Kcna4 50 * Kcna5 5HT Kcna6 In situ hybridization for Avpr1a, tryptophan hydroxylase, and c-fos mRNA (a gene up- e (mV) Kcna7 d 40 Kcnb1 regulated by neuronal activity) revealed that, in both male and female subjects, expo- u Kcnb2 t i Kcnc1 High l Kcnc2 Avpr1a sure to a female stimulus animal increased c-fos expression in the DR both in p 30 Kcnc3 Kcnc4 m Tph2 Kcnd1 Serotonin Avpr1a+ neurons and non-Avpr1a neurons. A 20 Action Potential Kcnd2 Kcnd3 Phenotype Slc6a4 0 mV Kcnf1 10 Kcng1 Kcng3 GABA Gad1 Our data verify the existence of AVP-responsive neurons in the DR and suggest that Kcng4 Kcnh1 Phenotype Gad2 activation of Avpr1a+ neurons in the DR is stimulus specific, and may be initiated by Pet1 Drd2/Pet1 Avpr1a Kcnh2 Avpr1a-GFP neurons had... Kcnh4 Kcnh5 Glutamate Slc17a6 more affiliative, less antagonistic interactions. * smaller amplitude APs Kcnh6 3.0 10 mV Kcnh7 Phenotype Low Kcnh8 Slc17a8 * shorter duration APs Kcnj1 2.5 Kcnj10 Ar Avpr1a neurons have distinct physiological properties compared to neighboring sero- 10 ms * presence of depolarizing Kcnj11 Kcnj12 Kcnj13 Esr1 tonin neurons. Avpr1a neurons have small amplitude short duration action potentials. 2.0 after-potential (DAP) Kcnj15 Hormone n (ms) Kcnj16 During the after-hyperpolarization Avpr1a neurons have depolarizing after potential. o Esr2 i Kcnj2 Receptors t 1.5 Kcnj3 These properties likely facilitate the high firing rates observed in these neurons. a * Kcnj5 Pgr r Kcnj6 u Kcnj9 D 1.0 Kcnk1 Nr3c1 Action Potential Kcnk10 Kcnk12 Preliminary gene expression studies suggest a number of candidate ion channels Kcnk15 Glial Marker Gfap 0.5 Kcnk18 that may underlie the hyper-excitability profile of dorsal raphe Avpr1a neurons. Kcnk2 Kcnk3 Kcnk4 Kcnk5 Pet1 Drd2/Pet1 Avpr1a -60 mV Kcnk6 Further study is needed to fully interrogate the function of AVP pathways, but one Kcnk7 Kcnk9 Kcnma1 * Preliminary analyses suggest that Avpr1a-GFP neurons may function of the BNST/MeA system appears to be activation of the neurons in the DR Traces at 180 pA stimulation (below) highlight the differences in action potential shape Kcnn1 Kcnn2 consist of several neurochemical subtypes with potential excitatory influence on serotonin neurons. and difference in excitability Kcnn3 Kcnn4 Kcnq2 Stimulus - 180 pA Stimulus - 180 pA Kcnq3 * Numerous candidate ion channels have been identified and Kcnq4 Kcnq5 more in depth analyses are underway to identify how these Future work will focus on validating expression of specific ion channel genes and Kcns1 Kcns2 channels might impact Avpr1a-GFP neuron excitability seek to identify the functional role of dorsal raphe Avpr1a neurons. Kcns3 Kcnt1 Kcnt2 Kcnu1 Kcnv1 Nalcn References Scn11a Scn1a Scn3a Scn4a Ho et al. (2010) Hormones and Behavior, 58(3):368-77 * Scn5a High Relative expression levels of ion channels in pooled Described Fos expression in BNST vasopressin neurons after mating in mice Scn8a Scn9a samples of Avpr1a-GFP neurons collected using flu- Avpr1a-GFP Non-Drd2/5-HT Each column orescence assisted cell sorting (FACS) indicate Niederkofler et al. (2016) Cell Reports, 17(8):1934-49 represents a likely candidates for future exploration. First described the Drd2-expressing subset of serotonin neurons The delay in AP firing indicated by * in the serotonin neuron indicates additional different mouse Low currents may slow firing in the subtype Rood and De Vries (2011) Journal of comparative Neurology, 519(12):2434-74 Avpr1a-GFP Presented a comprehensive atlas of the mouse vasopressin system Serotonin (TPH+) Rood and Beck (2014) Neuroscience, 260:205-16 First demonstrated interaction between vasopressin and serotonin neurons in the dorsal raphe Patched Avpr1a-GFP Work presented in this poster was supported by the NIMH through grants F32 MH096393 and K01 MH109712 awarded to B.D.R. Gong et al. (2003) Nature, 425(6961):917-25 Avpr1a-GFP mice were generated through the GENSAT program and obtained form the Mutant Mouse Resource & Research Center.
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