Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: A) Immunocytochemistry images of bovine sperm, human placenta, bovine placenta, and mouse brain depicting nuclear staining with DAPI (blue), CB1 (red), and merged images. FITC-PSA fluorescence can be observed in the merged image of sperm (green), for identification of the sperm acrosome. B) Western blot image of bovine sperm and equine pituitary identifying a single and specific band at the expected 75 kD value. Scale bar = 50 μM.

Article Snippet: Protein detection was performed with Anti-CB1(1:1500 proteintech Cat No: 17978-1-AP) in TBST buffer overnight at 4°C followed by detection with anti-rabbit secondary antibody HRP-linked (1:2000; Cat. #7074P2, Cell Signaling Technology ® ), in TBST buffer.

Techniques: Immunocytochemistry, Staining, Fluorescence, Western Blot

Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: Localization of CB1 in non-capacitated sperm (upper panel), to the post-acrosomal sheath as compared to loss of abundance when sperm are held in capacitating conditions (middle panel) as indicted by tyrosine phosphorylation staining (YPO 3-, yellow). Sperm exposed to THC under non-capacitating conditions exhibit decreased YPO 3 , CB1 and acrosome detection (green) (lower panel). Scale bar = 50 μm.

Article Snippet: Protein detection was performed with Anti-CB1(1:1500 proteintech Cat No: 17978-1-AP) in TBST buffer overnight at 4°C followed by detection with anti-rabbit secondary antibody HRP-linked (1:2000; Cat. #7074P2, Cell Signaling Technology ® ), in TBST buffer.

Techniques: Phospho-proteomics, Staining

Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: A) Immunocytochemistry images of bovine sperm, human placenta, bovine placenta, and mouse brain depicting nuclear staining with DAPI (blue), CB1 (red), and merged images. FITC-PSA fluorescence can be observed in the merged image of sperm (green), for identification of the sperm acrosome. B) Western blot image of bovine sperm and equine pituitary identifying a single and specific band at the expected 75 kD value. Scale bar = 50 μM.

Article Snippet: Primary antibody incubation for detection of CB1 include the addition of 50 μL of Anti-CB1(proteintech ® Cat No: 17978-1-AP) at a dilution of 1:50 in BB for 15 h at 4 ° C. Slides were then washed and 100μL of secondary antibody (Donkey-AntiRabbit Abcam Cat No: ab6799) was added at a dilution of 1:100 (in BB) for 1 h in the dark followed by washing.

Techniques: Immunocytochemistry, Staining, Fluorescence, Western Blot

Journal: bioRxiv

Article Title: Tetrahydrocannabinol exposure to postejaculatory sperm compromises sperm structure, function, the epigenome, and early embryo development

doi: 10.64898/2026.03.23.713385

Figure Lengend Snippet: Localization of CB1 in non-capacitated sperm (upper panel), to the post-acrosomal sheath as compared to loss of abundance when sperm are held in capacitating conditions (middle panel) as indicted by tyrosine phosphorylation staining (YPO 3-, yellow). Sperm exposed to THC under non-capacitating conditions exhibit decreased YPO 3 , CB1 and acrosome detection (green) (lower panel). Scale bar = 50 μm.

Article Snippet: Primary antibody incubation for detection of CB1 include the addition of 50 μL of Anti-CB1(proteintech ® Cat No: 17978-1-AP) at a dilution of 1:50 in BB for 15 h at 4 ° C. Slides were then washed and 100μL of secondary antibody (Donkey-AntiRabbit Abcam Cat No: ab6799) was added at a dilution of 1:100 (in BB) for 1 h in the dark followed by washing.

Techniques: Phospho-proteomics, Staining

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative Pinceaux staining showing a PC Soma (green, calbindin) along with the characteristic pinceaux formation on the axon side (red, CB1R) in both genotypes. B. Quantification of average CB1R pinceaux intensity across both genotypes and slide sets (VGAT, VGLUT1, VGLUT2). C. Same as B, but area of pinceaux. D-F. Proportion of CB1R puncta that colocalize with the respective vesicular marker before and after a 100 pixel shift of the CB1R channel along the X axis.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining, Marker

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative confocal images from WT, DMD mdx and CB1R KO cerebella demonstrating calbindin (green), raw CB1R (red), and filtered CB1R puncta (see methods: image analysis). The right column shows an expanded and merged view of the area indicated in the filtered CB1R column. Quantification of mean CB1R puncta intensity ( B ), density ( C ), and area ( D ) in WT (black) and DMDmdx (blue) cerebella.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques:

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGAT (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGAT image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGAT puncta in the molecular layer. E. Percentage of VGAT puncta colocalized with CB1R puncta. F. Intensity of CB1R puncta colocalized with VGAT puncta. G. Intensity of CB1R puncta not colocalized with VGAT puncta.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGLUT1 (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGLUT1 image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGLUT1 puncta in the molecular layer. E. Percentage of VGLUT1 puncta associated with CB1R puncta. F. Intensity of CB1R puncta colocalized with VGLUT1 puncta. G. Intensity of CB1R puncta not colocalized with VGLUT1 puncta

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGLUT2 (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGLUT2 image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGLUT2 puncta in the molecular layer. E. Percentage of VGLUT2 puncta associated with CB1R puncta. F. Average intensity of CB1R puncta colocalized with VGLUT2 puncta. G. Intensity of CB1R puncta not colocalized with VGLUT2 puncta.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: Molecular Medicine

Article Title: Progressive endocannabinoid system dysregulation in autosomal dominant polycystic kidney disease

doi: 10.1186/s10020-026-01457-w

Figure Lengend Snippet: ECS components are progressively dysregulated in human ADPKD kidney tissue. a Microarray analysis ( GSE7869 ) of human kidney tissue shows stepwise increases in CNR1 transcript from healthy cortex to minimally cystic (PKDm) and fully cystic (PKD) ADPKD tissue, with corresponding reductions in AEA-metabolizing enzymes NAPEPLD and FAAH . b Single-nucleus RNA-sequencing (snRNA-seq) analysis of human ADPKD kidneys ( n = 8) versus healthy controls ( n = 5) demonstrate consistent CNR1 upregulation and marked downregulation of NAPEPLD and FAAH , while 2-AG-metabolizing enzymes remain largely unchanged. c snRNA-seq analysis of diabetic kidney disease (DKD; n = 5 patients; controls n = 6) reveals minimal alterations in CNR1 and ECS-metabolizing enzymes. d Gene expression analysis by qPCR confirms CNR1 upregulation in human ADPKD kidney tissue ( n = 17) versus non-cystic nephrectomy controls ( n = 5), with concurrent changes in ECS enzyme transcription. e-i eCB quantification by liquid chromatography-tandem mass spectrometry (LC–MS/MS) reveals significant depletion of tissue anandamide (AEA); e , N -oleoylethanolamine (OEA); f , 2-arachidonoylglycerol (2-AG); h , and arachidonic acid (AA); i , while N -palmitoylethanolamine (PEA); g remains unchanged. j-l Western blot analysis shows substantial inter-individual variability in CB 1 R protein levels without significant difference between ADPKD ( n = 6) and control kidneys ( n = 4), but significant reductions in DAGLα/β, NAPEPLD, MGLL and FAAH protein. Western blots were normalized to total proteins. Data represent mean ± SEM. Statistics of control versus PKD represented by * and control versus PKDm by #. Statistical significance assessed by Mann–Whitney U test or unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Sections were stained with rabbit anti-CB 1 R antibody (1:200, 10006590, Cayman) followed by a goat anti-rabbit HRP conjugate (ImmPRESSTM, Vector laboratories).

Techniques: Microarray, RNA Sequencing, Gene Expression, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Western Blot, Control, MANN-WHITNEY

Journal: Molecular Medicine

Article Title: Progressive endocannabinoid system dysregulation in autosomal dominant polycystic kidney disease

doi: 10.1186/s10020-026-01457-w

Figure Lengend Snippet: Temporal endocannabinoid system dysregulation during Pkd1 RC/RC disease progression. a-g Quantitative PCR analysis of ECS-related genes across disease stages (9 and 12 months) showing progressive upregulation of Cnr1 a and Cnr2 b transcripts, with corresponding changes in ECS metabolic enzymes: Dagla c , Daglb d , Napepld e , Mgll f , and Faah g . h-n Western blot analysis and quantification of ECS proteins across disease stages, demonstrating sustained CB 1 R protein elevation at 9 and 12 months h, i , stable 2-AG-related enzyme proteins (DAGLα, DAGLβ, MGLL; h, j, k, m ), and stage-specific changes in AEA-related enzymes including elevated NAPEPLD at 12 months ( h, l ) and reduced FAAH at 9 months ( h, n ). Data represent mean ± SEM from WT ( n = 5) and Pkd1 RC/RC ( n = 6–7) per group per timepoint. Statistical analysis: unpaired t -test comparing Pkd1 RC/RC to age-matched WT. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus WT

Article Snippet: Sections were stained with rabbit anti-CB 1 R antibody (1:200, 10006590, Cayman) followed by a goat anti-rabbit HRP conjugate (ImmPRESSTM, Vector laboratories).

Techniques: Biomarker Discovery, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Nutrition

Article Title: Cannabis oil modulates liver alterations and endocannabinoid system changes in a female rat model of diet-induced MASLD

doi: 10.3389/fnut.2026.1770150

Figure Lengend Snippet: Endocannabinoids serum levels and liver CB1 protein mass in female rats fed a reference diet (RD), sucrose-rich diet (SRD) or SRD with cannabis oil (SRD + Ca). (A) Serum anandamide (AEA) levels. (B) Serum 2-arachidonoylglycerol (2-AG) levels. (C) Liver protein mass levels of CB1. Each gel contained an equal number of samples from rats fed a RD, SRD and SRD + Ca. Upper. Representatives immunoblot of liver CB1. Bottom. Densitometric immunoblot analysis of the CB1 protein mass levels. Data are expressed as mean ± SEM ( n = 6). Statistical differences were evaluated by one-way ANOVA followed by the Newman–Keuls post-hoc test (* P < 0.05 and # P < 0.05 vs. SRD and RD).

Article Snippet: The membranes were probed with mouse primary monoclonal antibodies against CB1 (mouse monoclonal antibody; sc-293419; Santa Cruz Biotecnology) and then incubated with goat anti-mouse IgG conjugated to horseradish peroxidase antibody (mIgG-Fc-BP-HRP; sc-525409, Santa Cruz Biotechnology).

Techniques: Cannabis, Western Blot