Why use the NTG model instead of just CGRP?

NTG acts as a nitric oxide donor that activates the entire trigeminovascular pathway, replicating more clinical features (including nausea-like behaviors) compared to the more targeted CGRP induced model.

Can you measure photophobia in mice?

Yes. We use a standardized Light/Dark box assay where we measure the time spent in the dark chamber and the number of transitions following an NTG challenge.

Is orofacial testing necessary?

Yes. While hind-paw allodynia occurs in migraine models (extracephalic allodynia), orofacial (cephalic) allodynia is the primary clinical symptom and provides higher translational validity for migraine drugs.

How do you differentiate between the effects of acute (abortive) vs. preventative treatments in your models?

Our platform offers flexible modeling windows tailored to the drug's clinical intent: Acute (Abortive) studies measure the reversal of established allodynia by administering compounds after a single provocation (e.g., NTG), while Preventative (Prophylactic) studies utilize chronic, repetitive provocation protocols (e.g., NTG every other day for 9 days) to evaluate the prevention of sensitization over time.

What is the significance of measuring CSD?

CSD is the electrophysiological substrate of the migraine aura. It is a wave of profound neuronal depolarization that spreads across the cortex. Measuring CSD frequency and velocity is essential for candidates targeting the early initiation phase of migraine or drugs specifically intended for "Migraine with Aura."

How do you ensure that behavioral results (like photophobia) aren't just caused by general sedation?

Every study can include an Open Field Test (OFT) or Rotarod assessment. If a compound reduces photophobia or allodynia without reducing the animal's total distance traveled or motor coordination, we can confidently conclude the effect is analgesic/anti-migraine rather than sedative.

Can you detect CGRP levels in small volume samples from mice?

Yes. By utilizing the Meso Scale Discovery (MSD) or SIMOA platforms, we can quantify CGRP and PACAP-38 in volumes as low as 25 µL. This allows us to perform serial blood sampling or dural tissue analysis even in mouse models with high sensitivity (femtogram/mL range).

What positive controls do you typically use?

We utilize a range of clinically relevant positive controls tailored to the study's mechanism of action, including Sumatriptan for acute rescue models, Olcegepant/Ubrogepant for CGRP-targeted validation, and Topiramate or Propranolol for chronic and preventative efficacy studies.

Migraine Modeling & Pharmacodynamics Services

Introduction

Migraine is a debilitating neurovascular disorder primarily driven by trigeminovascular activation, neuropeptide dysregulation (notably Calcitonin gene-related peptide (CGRP)), and cortical spreading depression (CSD), often presenting with high-prevalence comorbidities such as photophobia and nausea. As a leading cause of global disability, migraine affects approximately 15% of the world's population, with a significantly higher prevalence in women and strong associations with obesity and depression. To address the complexities of migraine chronification and the high burden of associated symptoms, Creative Biolabs' platform offers a comprehensive suite of high-fidelity models, including the "Gold Standard" Nitroglycerin (NTG) model, the CSD aura model, and the electrical stimulation of the trigeminal ganglion model, to capture every pathological phase of the disease. By integrating advanced pharmacodynamic readouts such as orofacial Von Frey testing, photophobia assessment, and laser speckle contrast imaging, we provide the robust, clinically translatable data necessary to evaluate next-generation acute rescue drugs and prophylactic therapies targeting dopaminergic, serotonergic, and CGRP pathways.

Fig.1 This schematic highlights the integration of ascending/descending pathways, trigeminocervical convergence, and autonomic reflexes in migraine pathophysiology. (OA Literature) Fig.1 This schematic illustrates the complex interplay between ascending and descending pathways, the convergence of dural and cervical inputs within the trigeminocervical complex, and the integration of the trigeminal autonomic reflex in migraine pathophysiology.¹

Available Migraine Models

Our platform offers a specialized suite of migraine models designed to replicate the complex neurovascular mechanisms of the disease. These models support the evaluation of acute abortive treatments (e.g., Triptans, Gepants) and preventative therapies (e.g., CGRP antibodies).

Migraine Models	Modeling Methods	Application Values	Animal Species
Nitroglycerin (NTG) induced Model of Migraine	Systemic administration of NTG (an NO donor): Acute: Single injection. Chronic: Intermittent dosing (5 doses over 9 days).	Pharmacological Gold Standard. Triggers predictable, delayed periorbital and hind-paw allodynia. Mimics spontaneous attacks and central sensitization. Ideal for testing acute "abortive" treatments (e.g., Triptans, CGRP antagonists) and preventative drugs (e.g., Topiramate, Monoclonal Antibodies).	Mouse, Rat
Cortical Spreading Depression (CSD) Model of Migraine Aura	Cortical stimulation via chemical (KCl), mechanical (pinprick), or electrical means to induce a slowly propagating wave of neuronal depolarization.	Primary paradigm for Migraine with Aura. Focuses on the neurological phenomenon preceding the headache. Mimics the physiological basis of aura and subsequent trigeminovascular activation. Robust platform for investigating cortical triggers and cortical excitability modulators.	Mouse, Rat
Electrical Stimulation of Trigeminal Ganglion Model	Stereotaxic microelectrode stimulation of the Trigeminal Ganglion (TG) to deliver high-intensity pulses, directly activating the trigeminovascular system.	Model for Neurogenic Inflammation. Triggers the release of CGRP and substance P, causing dural vasodilation and plasma protein extravasation. The gold standard for studying vascular activation and evaluating the efficacy of triptans and CGRP-targeted therapies (e.g., Aimovig or Ubrelvy).	Mouse, Rat

Evaluation Platform

Our platform utilizes sensitive behavioral and physiological readouts to quantify migraine symptoms. Our platform integrates peripheral nociception, affective phenotyping, and advanced neuroimaging to provide a comprehensive pharmacodynamic (PD) profile for novel anti-migraine therapeutics.

Behavioral Assessment: Behavioral readouts are the primary indicators of "headache" and "allodynia" in rodents.
- Mechanical Allodynia: Measured using von Frey filaments on the periorbital (facial) and plantar (paw) regions to detect hypersensitivity.
- Thermal/Cold Hyperalgesia: Using the Hargreaves test or Acetone test to measure reaction latency to temperature changes.
- Photophobia & Phonophobia: Assessed via the Light/Dark Box test (time spent in the dark) and noise induced grooming behavior.
- Home-Cage Activity: Using automated tracking (e.g., Any-maze) to observe reduced locomotion, mimicking the "desire to rest" during a migraine.
Electrophysiological & Imaging: These techniques monitor the "brain events" (e.g., CSD) and vascular changes in real-time.
- DC Potential Recording: Monitoring the negative shift (typically -15 to -30 mV) during a CSD wave.
- Electrocorticography (ECoG): Measuring the depression of cortical electrical activity following a depolarization wave.
- Laser Doppler Flowmetry (LDF): Real-time monitoring of dural blood flow and vasodilation, essential for testing vasoactive drugs like Triptans.
- Intrinsic Optical Imaging (IOI): Visualizing the slow propagation of CSD waves across the cortex.
Biochemical & Molecular Analysis: Measuring the "chemical signatures" of migraine to confirm target engagement.
- CGRP Measurement: Quantifying CGRP levels in the jugular vein or cerebrospinal fluid (CSF) via ELISA or Radioimmunoassay (RIA).
- c-Fos Mapping: Using immunohistochemistry to detect neuronal activation in the Trigeminal Nucleus Caudalis (TNC).
- Neuroinflammation Markers: Measuring cytokines (IL-1β, TNF-α) and neuropeptides (Substance P) in the dura mater or trigeminal ganglion.
- NO/cGMP Pathway: Measuring cyclic GMP levels to validate the mechanism of Nitroglycerin induced models.

Applications

Acute Rescue Therapies: Testing "abortive" drugs (e.g., Gepants, Triptans) in the NTG or CGRP induced hyperalgesia models.
Prophylactic Treatments: Evaluating long-term preventative agents (e.g., Monoclonal antibodies) in chronic IS or NTG models.
CSD Inhibitors: Screening candidates intended to block the aura phase of migraine.
Mechanism of Action (MoA) Studies: Investigating the role of ion channels (e.g., KATP, P2X3) in trigeminal signaling.

Our Advantages

Superior Translational Relevance: We move beyond standard hind-paw testing by utilizing the Orofacial Von Frey system to monitor periorbital and snout hypersensitivity. This accurately replicates the cephalic sensitization characteristic of clinical migraine, providing higher predictive value for clinical success.
Objective Multi-Symptom Phenotyping: Our platform integrates AI-driven Grimace Scales to evaluate spontaneous pain and automated Light/Dark boxes to quantify photophobia. All experiments are conducted in sound-attenuated, light-controlled environments to ensure highly stable and reproducible baselines.
Real-Time Neurovascular Imaging: We employ Laser Speckle Contrast Imaging (LSCI) and high-throughput intrinsic optical imaging to quantitatively monitor dural vasodilation and CSD waves in real-time, providing direct visual evidence of drug effects on neurovascular coupling.
Translational Molecular Data Chain: Leveraging the MSD/SIMOA platforms, we detect femtogram-level release of CGRP and Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP)-38. These biomarkers are correlated with c-Fos neural mapping in the TNC to achieve full-chain validation from the periphery to the central nervous system.
Migraine Chronification Modeling: We offer specialized repetitive induction protocols (using NTG or "Inflammatory Soup") to simulate the transition from episodic to chronic migraine. This provides a rigorous environment for validating long-term preventative therapies, such as monoclonal antibodies.

Work with Us

Inquiry Stage:

Summarize the project requirements and fill in the information collection form.
Sign a CDA from both parties to further communicate information, such as targets.
Select an animal model, discuss experimental design, and determine assay parameters.
Project costing and project schedule forecasting.

Project Start:

We provide a detailed project plan, including the required sample quantities, methods, and protocols.
Both parties confirm the project details and start the project.
Confirm the timeline of the project.

Project Progress:

We provide periodic results and information on the animal's condition.
We will work together to make project adjustments as necessary.

Project Completion:

We provide a comprehensive project report promptly.
We arrange transportation for the produced samples.
We provide a discussion of the project results and help to arrange the next steps.

After-Sales Support:

Data storage and archiving.

Get in touch!

FAQs

Q: Why use the NTG model instead of just CGRP?

A: NTG acts as a nitric oxide donor that activates the entire trigeminovascular pathway, replicating more clinical features (including nausea-like behaviors) compared to the more targeted CGRP induced model.
Q: Can you measure photophobia in mice?

A: Yes. We use a standardized Light/Dark box assay where we measure the time spent in the dark chamber and the number of transitions following an NTG challenge.
Q: Is orofacial testing necessary?

A: Yes. While hind-paw allodynia occurs in migraine models (extracephalic allodynia), orofacial (cephalic) allodynia is the primary clinical symptom and provides higher translational validity for migraine drugs.
Q: How do you differentiate between the effects of acute (abortive) vs. preventative treatments in your models?

A: Our platform offers flexible modeling windows tailored to the drug's clinical intent: Acute (Abortive) studies measure the reversal of established allodynia by administering compounds after a single provocation (e.g., NTG), while Preventative (Prophylactic) studies utilize chronic, repetitive provocation protocols (e.g., NTG every other day for 9 days) to evaluate the prevention of sensitization over time.
Q: What is the significance of measuring CSD?

A: CSD is the electrophysiological substrate of the migraine aura. It is a wave of profound neuronal depolarization that spreads across the cortex. Measuring CSD frequency and velocity is essential for candidates targeting the early initiation phase of migraine or drugs specifically intended for "Migraine with Aura."
Q: How do you ensure that behavioral results (like photophobia) aren't just caused by general sedation?

A: Every study can include an Open Field Test (OFT) or Rotarod assessment. If a compound reduces photophobia or allodynia without reducing the animal's total distance traveled or motor coordination, we can confidently conclude the effect is analgesic/anti-migraine rather than sedative.
Q: Can you detect CGRP levels in small volume samples from mice?

A: Yes. By utilizing the Meso Scale Discovery (MSD) or SIMOA platforms, we can quantify CGRP and PACAP-38 in volumes as low as 25 µL. This allows us to perform serial blood sampling or dural tissue analysis even in mouse models with high sensitivity (femtogram/mL range).
Q: What positive controls do you typically use?

A: We utilize a range of clinically relevant positive controls tailored to the study's mechanism of action, including Sumatriptan for acute rescue models, Olcegepant/Ubrogepant for CGRP-targeted validation, and Topiramate or Propranolol for chronic and preventative efficacy studies.

Published Data

The PACAP antagonist PACAP6-38 alleviates central sensitization in NTG induced chronic migraine rats by downregulating abnormally elevated synaptic-associated proteins in the TNC. Mechanistically, it repairs disrupted synaptic ultrastructures, restores aberrant dendritic morphology, and reverses impairments in synaptic plasticity through the inhibition of the ERK/CREB/BDNF signaling pathway.

Fig.2 The therapeutic effect of PACAP6-38 in NTG induced chronic migraine rats. (OA Literature) Fig. 2 The molecular mechanism of PACAP6-38 in the NTG induced chronic migraine rat model.²

References

Villar-Martinez, Maria Dolores, and Peter J Goadsby. "Pathophysiology and Therapy of Associated Features of Migraine." Cells vol. 11,17 2767. Distributed under Open Access license CC BY 4.0, without modification. https://doi.org/10.3390/cells11172767
Zhang, Lily et al. "PACAP6-38 improves nitroglycerin induced central sensitization by modulating synaptic plasticity at the trigeminal nucleus caudalis in a male rat model of chronic migraine." The Journal of Headache and Pain vol. 24,1 66. Distributed under Open Access license CC BY 4.0, with modification. https://doi.org/10.1186/s10194-023-01603-3