Ai32 mice

Manufactured by Jackson ImmunoResearch

Sourced in United States

The Ai32 mice are a strain of laboratory mice produced by Jackson ImmunoResearch. They are genetically engineered to express the Ai32 reporter gene, which allows for the visualization of specific cell types or neural circuits through the expression of a fluorescent protein.

Automatically generated - may contain errors

Lab products found in correlation

21 protocols using ai32 mice

Generation and Characterization of Piezo2 Knockout Mice

Cited 4 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Generation of the Advillin-CreERT2 Piezo2 conditional knockout and HoxB8-Cre Piezo2 knockout mice have been previously described (14 (link), 15 (link)). For both Cre-lines, Piezo2^fl/- mice containing one floxed allele and one null allele were used as Piezo2 knockouts, and littermates which did not carry the Cre allele and were homozygous or heterozygous for the floxed allele were used as controls. CGRP-GFP and MRGPRD-eGFP (25 (link)) mice were a generous gift from Drs. David Ginty and Mark Zylka, respectively. The Piezo2-GFP-IRES-Cre mouse was generated as previously described (21 (link)).
Piezo2-GFP-IRES-Cre mice were crossed to Ai32 mice (The Jackson Laboratory, (22 (link))) that carry the ChR2(H134R)-EYFP construct separated from its CAG promoter by a floxed stop sequence, ensuring the selective expression in Cre-positive tissues. Regarding the viral approach, Piezo2-GFP-IRES-Cre mouse pups (P0-P2) were intraperitoneally injected with AAV8-CAG-Flex-ChR2-tdtomato virus (10 ul per pup, virus titer 2×10¹³ GC/ml). Behavioral assessments were conducted at least 4 weeks post-injection.

Murthy S.E., Loud M.C., Daou I., Marshall K.L., Schwaller F., Kühnemund J., Francisco A.G., Keenan W.T., Dubin A.E., Lewin G.R, & Patapoutian A. (2018). The mechanosensitive ion channel Piezo2 mediates sensitivity to mechanical pain in mice. Science translational medicine, 10(462), eaat9897.

+ Open protocol

+ Expand

Optogenetic Manipulation of Nav1.8+ Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Nav1.8^ChR2 mice were generated by crossing Nav1.8^cre mice with Ai32 (RCL-ChR2(H134R)/EYFP) mice. Nav1.8^cre mice were gifts from Dr. John Wood at University College London and transferred to us from Dr. Stephen Waxman’s lab at Yale University. Ai32 mice were purchased from Jackson Labs. Animal care and use conformed to NIH guidelines for care and use of experimental animals. Experimental protocols were approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Alabama at Birmingham.

Yamada A., Yamada A.I., Ling J., Furue H., Luo W, & Gu J.G. (2023). Properties of Nav1.8ChR2-positive and Nav1.8ChR2-negative afferent mechanoreceptors in the hindpaw glabrous skin of mice. Molecular Brain, 16, 27.

+ Open protocol

+ Expand

Optogenetic Manipulations of Vglut2+ Neurons

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ten mice were used. We used Vglut2-Cre mice [Jackson laboratories, #028863, Vglut2-ires-cre knock-in (C57BL/6J); Vong et al., 2011 (link); Figures 1A,B], ChR2-EYFP-lox mice (Ai32 mice, Jackson laboratory, #024109, B6.Cg-Gt(ROSA)26Sor^{tm32(CAG-COP4*H134R/EYFP)Hze}/J; Madisen et al., 2012 (link); Figure 1B), and ZsGreen-lox mice (Ai6 mice, Jackson laboratory, #007906, B6.Cg-Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/J; Madisen et al., 2012 (link); Figure 1A). We crossed homozygous Vglut2-Cre mice with homozygous ChR2-EYFP-lox mice to obtain the double heterozygous Vglut2-ChR2-EYFP mice. We crossed homozygous Vglut2-Cre mice with homozygous ZsGreen-lox mice to obtain the double heterozygous Vglut2-ZsGreen mice. Animals had ad libitum access to food and water, with lights on from 6 AM to 8 PM. Mice were 16–36 weeks old for in vivo optogenetics (three males, two females), 10–18 weeks old for neuroanatomy (one male, two females), and 15–23 days old for patch-clamp experiments (one male, one undetermined).

van der Zouwen C.I., Boutin J., Fougère M., Flaive A., Vivancos M., Santuz A., Akay T., Sarret P, & Ryczko D. (2021). Freely Behaving Mice Can Brake and Turn During Optogenetic Stimulation of the Mesencephalic Locomotor Region. Frontiers in Neural Circuits, 15, 639900.

+ Open protocol

+ Expand

Optogenetic Activation of TRPV1+ Neurons

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

All procedures were approved by the Animal Care and Use Committee of Washington University and in strict accordance with the US National Institute of Health (NIH) Guide for the Care and Use of Laboratory Animals. Adult male and female mice (7-14 weeks of age) utilized in experiments were housed in Washington University School of Medicine animal facilities on a 12-hour light/dark cycle with access ad libitum to food and water.
For relevant optogenetic experiments, mice were generated with conditional expression of ChR2 in TrpV1 expressing sensory neurons by crossing heterozygous TrpV1-Cre mice (provided by Dr. Mark Hoon, NIDCR^{39 (link)}) with homozygous Ai32 mice (Stock 3: 012569, The Jackson Laboratory)^{40 (link)}. For the purposes of this study, we refer to these mice as “TRPV1:ChR2”. These mice were previously characterized in our lab^{41 (link)}. All other experiments were performed using C57BL/6J mice bred in house, originally obtained from The Jackson Laboratory.

McIlvried L.A., Del Rosario J.S., Pullen M.Y., Wangzhou A., Sheahan T.D., Shepherd A.J., Slivicki R.A., Lemen J.A., Price T.J., Copits B.A, & Gereau RW I.V. (2023). Intrinsic Homeostatic Plasticity in Mouse and Human Sensory Neurons. bioRxiv.

+ Open protocol

+ Expand

Optogenetic Manipulation of Sensory Neurons

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Adult mice (8–12 weeks of age) are utilized for this study. Mice are housed in the animal facilities of the Washington University School of Medicine on a 12 hour light/dark cycle, with access ad libitum to food and water. Institutionally approved protocols are followed for all aspects of this study.
Three Cre-driver lines are used for this study including heterozygous SNS-Cre mice from Rohini Kuner²¹, heterozygous TrpV1-Cre mice from Mark Hoon^{22 (link)}, and heterozygous Advillin-Cre mice provided by Fan Wang.^{14 (link)} Mice from each of these three lines are crossed to homozygous Ai32 mice from Jackson Laboratory. As previously described, Ai32 mice harbor ChR2 (H134R)-eYFP in the Gt(ROSA)26Sor locus.^{23 (link)} To generate mice with conditional expression of ChR2 in specific populations of sensory neurons, mice with ChR2 in the Rosa locus (Ai32 mice) are crossed to mice expressing cre from various sensory neuron-specific driver gene loci (Advillin, TrpV1, or SNS). For the purposes of this study, the three lines generated are referred to as Advillin-ChR2, TrpV1-ChR2, and SNS-ChR2, respectively.

Park S.I., Brenner D.S., Shin G., Morgan C.D., Copits B.A., Chung H.U., Pullen M.Y., Noh K.N., Davidson S., Oh S.J., Yoon J., Jang K.I., Samineni V.K., Norman M., Grajales-Reyes J.G., Vogt S.K., Sundaram S.S., Wilson K.M., Ha J.S., Xu R., Pan T., Kim T.I., Huang Y., Montana M.C., Golden J.P., Bruchas M.R., Gereau RW I.V, & Rogers J.A. (2015). Soft, stretchable, fully implantable miniaturized optoelectronic systems for wireless optogenetics. Nature biotechnology, 33(12), 1280-1286.

+ Open protocol

+ Expand

Genetic Labeling of Striatal Neuronal Inputs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Animal care and procedures used for these studies were approved by the Animal Care and Use Committee of the National Institute on Alcohol Abuse and Alcoholism and conformed to the guidelines of the US National Institutes of Health Guide for the Care and Use of Animals. For most experiments, 9–18 week old male C57Bl/6J mice (The Jackson Laboratory, stock no. 000664) were used. For experiments evaluating glutamatergic transmission in CINs, hemizygous ChAT-IRES-Cre mice (The Jackson Laboratory, stock no. 006410) were crossed with Ai14 mice (The Jackson Laboratory, stock no. 007908) to drive TdTomato reporter expression in CINs. For some experiments evaluating corticostriatal and thalamostriatal transmission, Ai32 mice (The Jackson Laboratory, stock no. 024109) were crossed with Emx1-IRES-Cre (The Jackson Laboratory, stock no. 005628) or vGlut2-IRES-Cre mice (The Jackson Laboratory, stock no. 016963, backcrossed at NIAAA to C57Bl/6J mice for at least six generations), respectively. This allowed Cre-dependent expression of ChR2 in the striatal inputs of interest. Animals were housed 2–4 per cage in a temperature- and humidity-controlled room with a standard 12 hour light/dark cycle and ad libitum access to food and water.

Johnson K.A., Mateo Y, & Lovinger D.M. (2017). Metabotropic glutamate receptor 2 inhibits thalamically-driven glutamate and dopamine release in the dorsal striatum. Neuropharmacology, 117, 114-123.

+ Open protocol

+ Expand

CaMKIIα-Cre Mice for Channelrhodopsin2 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Homozygous CaMKIIα-Cre line T29–1 transgenic mice (Jackson Laboratory #005359) were crossed with homozygous Ai32 mice (Jackson Laboratory #012569) to express channelrhodopsin2 (ChR2) in neurons expressing male and female CaMKIIα in F1 hybrid mice (N = 5; 3 male; 25–40g, 30–50 weeks of age). After implantation, animals were housed individually on a reversed 12/12 h day/night schedule. Following one week of recovery, mice were recorded 5–7 days/week for two months before being euthanized with pentobarbital cocktail (Euthasol®, transcardial 300 mg/kg) and perfused with formalin (10%). All experiments were conducted in accordance with the Institutional Animal Care and Use Committee of New York University Medical Center.

McKenzie S., Huszár R., English D.F., Kim K., Christensen F., Yoon E, & Buzsáki G. (2021). Pre-existing hippocampal network dynamics constrain optogenetically induced place fields. Neuron, 109(6), 1040-1054.e7.

+ Open protocol

+ Expand

Generating Mice with Optogenetic Control

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

All experimental procedures were approved by the committee for the Care and Use of Laboratory animals of the Government of Upper Bavaria, Germany. Mice selectively expressing ChR2(H134R)-EYFP in forebrain glutamatergic neurons were generated by breeding homozygous NEX-Cre mice (Goebbels et al., 2006 (link)) to homozygous Ai32 mice (Madisen et al., 2012 (link); purchased from the Jackson Laboratory). Genotyping was performed using the following primers specific for NEX-Cre: NEXCre4 5′-GAG-TCC-TGG-AAT-CAG-TCT-TTT-TC-3′, NEXCre5 5′-AGA-ATG-TGG-AGT-AGG-GTG-AC-3′, and NEXCre6 5′-CCG-CAT-AAC-CAG-TGA-AAC-AG-3′. Standard PCR conditions resulted in a Cre-specific PCR product of 525-bp and a wild-type PCR product of 770-bp. Genotyping for ChR2 was conducted according to the genotyping protocol provided by the Jackson Laboratory. All animals were housed under a 12 h light/dark cycle starting 5 days before the experiment and had access to water and food ad libitum. All experiments were performed in 8- to 12-week-old male mice.

Dine J., Genewsky A., Hladky F., Wotjak C.T., Deussing J.M., Zieglgänsberger W., Chen A, & Eder M. (2016). Local Optogenetic Induction of Fast (20–40 Hz) Pyramidal-Interneuron Network Oscillations in the In Vitro and In Vivo CA1 Hippocampus: Modulation by CRF and Enforcement of Perirhinal Theta Activity. Frontiers in Cellular Neuroscience, 10, 108.

+ Open protocol

+ Expand

Optogenetics in Mouse Somatosensory Cortex

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

This study is based on 61 transgenic mice heterozygous for Ntsr1-Cre (Strain: Tg(Ntsr1-cre)GN220Gsat/Mmucd). This strain expresses Cre recombinase in L6 CT projection cells (Bortone et al., 2014 (link); Gong et al., 2007 (link); Kim et al., 2014 (link)). All mice used had ICR genetic backgrounds (Charles River). Of the 61 mice used here, 50 were bred by crossing Ntsr1-Cre males with wild-type ICR female mice. These mice were used for the majority of our experiments following viral induction of ChR2 in the somatosensory CT pathway (below). The remaining mice were bred by crossing Ntsr1-Cre males with female mice homozygous for a conditional (Cre-dependent) ChR2-EYFP allele (Ai32 mice; Jackson Labs # 012569; n = 8 mice) or tdTomato allele (Ai9 mice; Jackson Labs; n = 3 mice). The Ntsr1/Ai32 mice were only used for the following experiments: 1) Low frequency (0.1 Hz) optical CT stimulation studies with paired ML electrical stimulation or, 2) studies involving electrical stimulation of TRN axons. All animals were group-housed, maintained on a 12:12 h light-dark cycle, and provided food and water ad libitum.

Crandall S.R., Cruikshank S.J, & Connors B.W. (2015). A corticothalamic switch: controlling the thalamus with dynamic synapses. Neuron, 86(3), 768-782.

+ Open protocol

+ Expand

Intracellular Recordings in Anesthetized Rodents

Check if the same lab product or an alternative is used in the 5 most similar protocols

All experiments were approved by the Institutional Animal Care and Use Committee (IACUC) at New York University Medical Center and the Ethics Committee of the Instituto Cajal (CSIC). Intracellular recordings in anesthetized rats were obtained in Madrid and were performed according to the Spanish legislation (R.D. 1201/2005 and L.32/2007), the European Communities Council Directives of 1986 (86/609/EEC) and 2003 (2003/65/CE) for animal research. All animals were kept in the vivarium on a 12-h light/dark cycle and were housed 2–3 per cage. Following surgery, the mice were moved to a 12-hour reverse light cycle (lights on/off at 7 pm/am) and housed individually. Prior to behavior training, mice were provided food and water ad libitum, but were water restricted to maintain 85% of their weight during and after behavioral training. Homozygous CaMKIIa-Cre line T29–1 transgenic mice (Jackson Laboratory #005359) were crossed with homozygous Ai32 mice (Jackson Laboratory #012569) to express channelrhodopsin2 (ChR2) in neurons expressing CaMKIIa in F1 hybrid mice. We used n = 7 male F1 hybrid mice (n = 3 mice for extracellular recordings and n = 4 for intracellular recordings in head-fixed conditions; 25–40 g, 30–50 weeks of age) and n = 10 male and female Wistar rats (240–450 gr; 20 to 50 weeks of age).

Valero M., Navas-Olive A., de la Prida L.M, & Buzsáki G. (2022). Inhibitory conductance controls place field dynamics in the hippocampus. Cell reports, 40(8), 111232.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!