Journal: Brain Structure & Function

Article Title: Transmitter and ion channel profiles of neurons in the primate abducens and trochlear nuclei

doi: 10.1007/s00429-021-02315-7

Figure Lengend Snippet: Summary of primary antibodies and dilutions for the immunolabeling

Article Snippet: Kv1.1 , Rabbit/polyclonal , Voltage-gated potassium channel 1.1 , Alomone Labs Jerusalem BioPark (JBP) , AB_2040144 , 1:250 (IF), 1:750 (IHC).

Techniques: Marker

Journal: Frontiers in Behavioral Neuroscience

Article Title: Alpha 1 -adrenergic receptor blockade in the ventral tegmental area attenuates acquisition of cocaine-induced pavlovian associative learning

doi: 10.3389/fnbeh.2022.969104

Figure Lengend Snippet: Effects of α 1 -AR blockade in the VTA on the acquisition of cocaine-evoked CPP. (A,F) The experimental time-line and schedule of intra-VTA microinfusions during the acquisition of CPP induced by one (A) or two (F) cocaine (20 mg/kg, i.p.) conditionings. (B) Graphical representation of the two methods of calculating conditioned responses in the CPP paradigm. CPP score is defined as a difference between time spent (t 2 ) in the cocaine-paired chamber minus time spent in (t 3 ) control chamber (saline-paired) during post-conditioning (post-test). Delta score is defined as a difference between time spent (t 2 ) in the cocaine-paired chamber during post-conditioning minus time spent (t 1 ) in the same chamber but during pre-conditioning (pre-test). (C,G) There were no pre-existing differences in the time spent in the cocaine- and saline-paired chambers during pre-test between future vehicle (Veh)- and prazosin (Praz)-treated subjects. (D,E) Intra-VTA microinfusion of prazosin attenuated the acquisition of CPP induced by one ( D for CPP score and E for delta score) and two ( H for CPP score and I for delta score) cocaine (20 mg/kg, i.p.) conditionings. (J) Localization of histologically verified cannula placements. Data are presented as individual data points as well as the mean and SEM. ** p < 0.01, *** p < 0.001, Newman–Keuls post-hoc test vs. Praz 0 μg.

Article Snippet: The specificity of anti-α 1 -AR antibody was tested by pre-incubation with manufacturer-supplied control antigen peptides (1:125, AK-490, Alomone Labs, Jerusalem, Israel) and comparing the strength of the signal between the control and blocked antibodies.

Techniques:

Journal: Frontiers in Behavioral Neuroscience

Article Title: Alpha 1 -adrenergic receptor blockade in the ventral tegmental area attenuates acquisition of cocaine-induced pavlovian associative learning

doi: 10.3389/fnbeh.2022.969104

Figure Lengend Snippet: Effects of α 1 -AR blockade in the VTA on cocaine reinforcement in the cocaine self-administration paradigm. (A) Rats, after intravenous catheter and intra-VTA cannula implantation surgeries and recovery were trained to self-administer cocaine (∼0.5 mg/kg/inf.) for 8 days. On day 9, intra-VTA vehicle (Veh) or prazosin (Praz) microinfusions were performed immediately prior to subsequent cocaine self-administration session. (B–D) There were no pre-existing differences during cocaine self-administration training in the number of (B) active and (C) inactive lever presses as well as in (D) the number of cocaine infusions between future vehicle (Veh)- and prazosin (Praz)-treated subjects. (E) Intra-VTA microinfusion of prazosin (Praz 1 μg/side) on day 9 had no effect on the numbers of active and inactive lever responses and on (F) the number of cocaine infusions. (G) Intra-VTA α 1 -AR blockade had no effects on the numbers of active lever presses over time. Similarly, prazosin treatment on day 9 had no effects on (H) the numbers of active and inactive lever presses and (I) cocaine infusions, 24 h after treatment (on day 10). (J) Finally, prazosin microinfusion on day 9 had no effects on active lever responding during time-out. (K) Localization of histologically verified cannula placements. Data are presented as individual data points as well as the mean and SEM. Act, active lever; Inact, inactive lever.

Article Snippet: The specificity of anti-α 1 -AR antibody was tested by pre-incubation with manufacturer-supplied control antigen peptides (1:125, AK-490, Alomone Labs, Jerusalem, Israel) and comparing the strength of the signal between the control and blocked antibodies.

Techniques:

Journal: Frontiers in Behavioral Neuroscience

Article Title: Alpha 1 -adrenergic receptor blockade in the ventral tegmental area attenuates acquisition of cocaine-induced pavlovian associative learning

doi: 10.3389/fnbeh.2022.969104

Figure Lengend Snippet: Effects of α 1 -AR blockade in the VTA on cocaine-evoked 50-kHz USVs. α 1 -AR blockade in the VTA with 1 μg/side prazosin (Praz) had no effect on the number of appetitive USVs after (A) saline (i.p.) or cocaine administration at (B) 10 mg/kg, i.p. or (C) 20 mg/kg dose. (D) Intra-VTA prazosin infusion had no effect on the cocaine-induced increase in the number of appetitive USVs and (E) did not modulate selected categories USVs after cocaine (20 mg/kg, i.p.) administration. (F) Localization of histologically verified cannula placements. Data are presented as individual data points as well as the mean and SEM. *** p < 0.001, Newman–Keuls post-hoc test vs. Cocaine 0 mg/kg.

Article Snippet: The specificity of anti-α 1 -AR antibody was tested by pre-incubation with manufacturer-supplied control antigen peptides (1:125, AK-490, Alomone Labs, Jerusalem, Israel) and comparing the strength of the signal between the control and blocked antibodies.

Techniques:

Journal: Frontiers in Behavioral Neuroscience

Article Title: Alpha 1 -adrenergic receptor blockade in the ventral tegmental area attenuates acquisition of cocaine-induced pavlovian associative learning

doi: 10.3389/fnbeh.2022.969104

Figure Lengend Snippet: Localization of α 1 -AR-expressing cells in the anterior parts of the VTA of TH-cre + rats. (A) Representative image of the EYFP- and α 1 -AR-expressing cells. (B–E) Magnified cells with α 1 -AR expression (red), EYFP (green), DAPI (blue), and merged channels showing α 1 -AR-immunoreactive and EYFP-expressing cells (white arrows) in the medial part of the anterior VTA. (F–I) Magnified cells with α 1 -AR (red), EYFP (green), DAPI (blue), and merged channels showing co-localized α 1 -AR-positive and EYFP-expressing cells (white arrows) in the lateral part of the anterior VTA. (J–Q) α 1 -AR expression in astrocytes (S100B) and GAD67-positive GABAergic interneurons. (J–M) Magnified images of astrocytes (red), α 1 -AR immunostaining (green), DAPI (blue), and composite images showing co-localization (white arrows) between α 1 -AR and S100B expression. (N–Q) Magnified images of GAD67-positive interneurons (red), α 1 -AR expression (green), DAPI (blue), and composite images showing co-localization between α 1 -AR and GAD67 immunoreactivity (white arrows). PBP, parabrachial pigmented nucleus; RLi, rostral linear nucleus.

Article Snippet: The specificity of anti-α 1 -AR antibody was tested by pre-incubation with manufacturer-supplied control antigen peptides (1:125, AK-490, Alomone Labs, Jerusalem, Israel) and comparing the strength of the signal between the control and blocked antibodies.

Techniques: Expressing, Immunostaining

Journal: Neurobiology of disease

Article Title: TAF1 -gene editing alters the morphology and function of the cerebellum

doi: 10.1016/j.nbd.2019.104539

Figure Lengend Snippet: List of primary and secondary antibodies used in the study.

Article Snippet: Low-Voltage-Activated CaV3.1 Calcium Channels , CaV3.1 , Rabbit , Almone labs , ACC-021 , 1:250 , .

Techniques: Binding Assay

Journal: Neurobiology of disease

Article Title: TAF1 -gene editing alters the morphology and function of the cerebellum

doi: 10.1016/j.nbd.2019.104539

Figure Lengend Snippet: (A) Representative photomicrographs of cerebellar slices from animals injected with (vehicle) or control or TAF1 gRNAs stained with an antibody against CaV3.1. Nuclei stained with DAPI. (B) Quantification of fluorescence intensity for the CaV3.1 immunohistochemistry. Scale bar = 200 μm. The experiments were performed in a blinded fashion. Data are shown as mean ± S.E.M. from 3 different fields from 3 animals per experimental condition (i.e. a total of nine fields per group). *p<0.05 versus; naïve, #p<0.05 versus gRNA-control (ANOVA followed by Tukey’s test). The experiments were conducted in an investigator blinded manner.

Article Snippet: Low-Voltage-Activated CaV3.1 Calcium Channels , CaV3.1 , Rabbit , Almone labs , ACC-021 , 1:250 , .

Techniques: Injection, Staining, Fluorescence, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Immunohistochemical analysis of ASIC1a expression in renal tissue. Immunostaining for ASIC1a, AQP1, THP and SYN in renal tissue. ASIC1a was colocalized with AQP and SYN, but not THP. ASIC1a, acid sensing ion channel 1a; AQP1, aquaporin 1, proximal tubular cells marker; THP, Tamm‐Horsfall protein, thick ascending limb and distal tubular cells marker; SYN, Synaptopodin, podocyte marker; DAPI, 4,6‐diamidino‐2‐phenylindole, nuclear

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Immunohistochemical staining, Expressing, Immunostaining, Marker

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Inhibiting ASIC1a by PcTx1 attenuate I/R induced kidney injury. Different doses of PcTx1 (0.2, 2, 4, 10 nmol L −1 kg −1 body weight) or vehicle were injected by tail vein, 30 min before I/R operation. Twenty‐four hours after reperfusion, the expression of ASIC1a in the kidney of sham and I/R operating mouse was measured by Western blotting and PCR; functional and histological changes were assessed by serum creatinine, Urea and PAS staining; apoptosis of renal tubular epithelia cells was assessed by TUNEL staining. A, I/R increase protein and RNA level of ASIC1a in the kidney, however, administration of PcTx1 had no effect on expression of ASIC1a. B‐C, injection of PcTx1 reduced serum creatinine and urea in a dose dependent manner; D, Typical visual field of PAS staining and pathological score calculated from PAS staining. PcTx1 (4 nmol L −1 kg −1 body weight) significantly attenuated I/R induced renal injury. * P < 0.05, ** P < 0.01, *** P < 0.001, compared with sham; # P < 0.05, ### P < 0.001, compared with vehicle+I/R, n = 6

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Injection, Expressing, Western Blot, Functional Assay, Staining, TUNEL Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Inhibiting ASIC1a by PcTx1 attenuate I/R induced apoptosis of renal tubule, in vivo. Typical image of TUNEL staining in renal coronal section from sham, vehicle+I/R and PcTx1+ I/R animal. *** P < 0.001, compared with sham; # P < 0.05, ### P < 0.001, compared with vehicle+I/R, n = 6

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: In Vivo, TUNEL Assay, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Inhibiting ASIC1a by PcTx1 reduced H/R induced apoptosis of HK‐2 cells. A, H/R increase protein level of ASIC1a in the HK‐2 cells. * P < 0.05, n = 6. B, HK‐2 cells were pre‐treated with different doses of PcTx1 (5, 25, 100, and 500 ng/mL) or vehicle before H/R treatment. Apoptosis of HK‐2 cells was measured by Annexin‐V/PI staining and evaluated by flow cytometry. C, the group data from B. PcTx1 attenuated H/R induced apoptosis especially early apoptosis dose dependently. *** P < 0.001, compared with Normoxia; # P < 0.05, ## P < 0.01, compared with vehicle+H/R, n = 6. E, Cell variability was measured by MTT assay. Treatment of PcTx1 for 6 h had no effect on variability of HK‐2 cells

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Staining, Flow Cytometry, MTT Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Inhibiting ASIC1a by PcTx1 reduced H/R induced intracellular calcium overload and loss of mitochondrial membrane potential. HK‐2 cells were administrated with different doses of PcTx1 as Figure . A, Intracellular calcium concentration were measured by fluo4 and evaluated by flow cytometry. PcTx1 attenuated H/R induced intracellular calcium overload dose dependently. B, Intracellular calcium concentration was measured by fluo4 immunofluorescence staining. As expected, PcTx1 attenuated H/R induced intracellular calcium overload. C, mitochondrial membrane potential were monitored by JC‐1 dye and evaluated by flow cytometry. *** P < 0.001, compared with Normoxia; # P < 0.05, ## P < 0.01, compared with vehicle+H/R, n = 6

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Concentration Assay, Flow Cytometry, Immunofluorescence, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Inhibiting ASIC1a by PcTx1 reduced expression of cleaved‐caspase3 in renal tubule. A: PcTx1 (4 nmol L −1 kg −1 body weight) or vehicle were injected by tail vein, 30 min before I/R operation. Twenty‐four hours after reperfusion, apoptosis of renal tubular epithelia cells was assessed by immunohistochemical reaction of cleaved‐caspase3. Typical image of cleaved‐caspase3 immunohistochemical staining in renal coronal section from sham, vehicle + I/R and PcTx1+ I/R animal; B, detailed information of immunohistochemical reaction of cleaved‐caspase3 under high‐power lens; ** P < 0.01, compared with sham; ## P < 0.001, compared with vehicle + I/R, n = 6. C, HK‐2 cells were pre‐treated with PcTx1 (100 ng/mL) before H/R treatment, cleaved‐caspase3 was measured by Western blotting; ** P < 0.01, compared with Normoxia; ## P < 0.01, compared with vehicle+H/R, n = 6

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Expressing, Injection, Immunohistochemical staining, Staining, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Acid‐sensing ion channel 1a is involved in ischaemia/reperfusion induced kidney injury by increasing renal epithelia cell apoptosis

doi: 10.1111/jcmm.14238

Figure Lengend Snippet: Schema of I/R induced the local microenvironment acidification and the activation of ASIC1a on renal injury and the involved underlying mechanism. Ischaemia caused accumulation of extracellular protons (H + ). ASIC1a are activated by extracellular H + and induce influx of Ca 2+ , which induced loss of mitochondrial membrane potential. The damage of mitochondrial increased cleaved‐caspase3, which resulted in apoptosis of renal tubular epithelia cells. Administration of the specific inhibitor of ASIC1a, PcTx1 ameliorated ischaemic renal injury by inhibiting ASIC1a, which implied a potential therapeutic choice for AKI

Article Snippet: After incubated in 0.3% BSA, the slices was incubated in the mixed primary antibodies: Guinea pig anti‐ASIC1a (1:50 alomone lab, Israel)+Rabbit anti‐aquaporin 1 (AQP1, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐ Tamm‐Horsfall protein (THP, 1:100, Abcam, UAS), Guinea pig anti‐ASIC1a (1:50)+Rabbit anti‐Synaptopodin (SYN, 1:100, Abcam, USA), Guinea pig anti‐ASIC2a (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC2a (1:50)+Rabbit anti‐SYN (1:100), Guinea pig anti‐ASIC3 (1:50 alomone lab, Israel)+Rabbit anti‐AQP1(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐THP(1:100), Guinea pig anti‐ASIC3 (1:50)+Rabbit anti‐ SYN (1:100).

Techniques: Activation Assay