• CaV3.2

    Raw current traces and current–voltage relationships of the CaV3.2 peak current recorded on the Port-a-Patch

CaV3.2 | voltage-dependent, T type, alpha 1H subunit calcium channel

Family:
Calcium channels

Subgroups:
L-Type (CaV1.1–CaV1.4), P/Q-Type (CaV2.1), N-Type (CaV2.2), R-Type (CaV2.3), T-Type (CaV3.1–CaV3.3)

Topology:
Six transmembrane alpha helices (S1–S6), total of four homologous domains make up the tetrameric alpha subunit structure

Assembling:
One large alpha subunit forms a functional channel, accessory subunits ( α1, α2δ, β1-4, and γ) are crucial for robust expression, they functionally modulate the alpha subunit

CaV3.2 Background Information


Overview:

CaV3.2 give rise to the T-type currents: Low-voltage-activated calcium channels are referred to as 'T' type because their currents are both transient, owing to fast inactivation, and tiny, owing to small conductance. T-type channels are thought to be involved in pacemaker activity, low-threshold calcium spikes, neuronal oscillations and resonance, and rebound burst firing. T-type channels serve pacemaking functions in both central neurons and cardiac nodal cells and support calcium signaling in secretory cells and vascular smooth muscle. They may also be involved in the modulation of firing patterns of neurons which is important for information processing as well as in cell growth processes.

In the adrenal zona glomerulosa, CaV3.2 participates in the signaling pathway leading to aldosterone production in response to either AGT/angiotensin II, or hyperkalemia. Furthermore, CaV3.2 has a proexcitatory impact in small-diameter nociceptors expressing mechanoactivated channels. It is expressed in primary sensory neurons of the dorsal root ganglion (DRG) and contributes to nociceptive and neuropathic pain. CaV3.2 participates in the development of inflammatory hyperalgesia and might play an important role in the sub-acute phase of inflammatory pain due to increased co-localization with TRPV1 receptors.


Data Sheet:

Gene:
CACNA1H

Human Protein:
UniProt O95180

Tissue:
Brain, ovary, placenta, vascular smooth muscle

Function/ Application:
Pacemaker activity (brain, heart), hormone secretion, fertilization

Pathology:
Angina, epilepsy (CAE6, EIG6), sleep, breast cancer, autism, pain, cardiac hypertrophy, Hyperaldosteronism, Familial, Type Iv

Interaction:
GNG2, KCNMA1, KDM5B, CACNA1s, CACNBs, CACNGs, NCAM

Modulator:
Kurtoxin, mibefradil, flunarizine, zonisamide, bepridil, nifedipine

Assays:
Patch Clamp: whole cell, room temperature

Particularities:
CaV channels often show a rundown phenomenon. Adequate intra- and extracellular solutions are essential for a good data quality.

Reviews and Links

Data and Applications

CaV3.2 - T-Type Calcium Channels

Cav32 IV

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx Millipore.

Shown are raw current traces (top left) and average peak current data (top right) of the current voltage relationship of CaV3.2 (T-type Ca2+- Channel) stably expressed in HEK293 cells. Activation and Inactivation plots were constructed (bottom). Half-activation and half-inactivating potentials were determined as -32 mV and -65 mV, respectively.

 

 

CaV3.2 - Raw Currents

SyncroCav32Currents

icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Cytomyx Millipore, UK.

Shown are raw current responses of CaV3.2 (HEK293) to a current voltage relationship step protocol. 75 % of the cells had a seal above 0.5 GΩ (color coded in green). The corresponding online analysis of this experiment is shown in the data set CaV3.2 - Online Analysis.

 

CaV3.2 - Mibefradil Antagonism

Cav32 Mibefradil

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

Dose dependent block by Mibefradil on current traces from an individual cell expressing CaV3.2. Data was averaged and fitted to the Hill equation

 

CaV3.2 - Inactivation

Cav32 inactivation

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

Current responses of a double pulse protocol with varying test potentials between the pulses (5 s) was used to determine the half inactivating potential. Peak current responses to the second pulse are expressed relative to the response to the first pulse. Both curves in Figure 5 were fitted to the Boltzmann equation and revealed a half-inactivating potential of -65 mV and a half-activating potential of -33 mV.

CaV3.2 - Current-to-Voltage Relationship

IV Cav32

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore

Representative current responses of an individual cell expressing Ca 2.2 to a standard voltage protocol. The average mean current at -20 mV of all recorded cells was -785 ± 110 pA (n = 12).

CaV3.2 - Channel Blockers

Cav32 Block

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx Millipore.

Shown are raw current traces (top) and average dose response curves (bottom) of CaV3.2 (T-type Ca2+- Channel) block by compounds as indicated. CaV3.2 is stably expressed in HEK293 cells. IC50s were 863 nM (Mibefradil), 27 μM (Nifedipine) and 52 μM Amiloride.

 

 

 

 

Application Notes

CaV3.2 - "High Throughput Pharmacology of CaV3.2 Channels on Nanion’s SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.  

CaV3.2 - "Characterization of CaV3.2 on Nanion's Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.7 MB)
Cells were kindly provided by Millipore.  

CaV3.2 - "Characterization of CaV3.2 (HEK293) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.

Webinars

28.04.2020 | Webinar: Validation and optimization of automated patch clamp voltage-gated Ca2+ channel assays

icon pl   Patchliner Webinar

Date: April 28. 2020, 4:00 PM CET (10:00 AM EDT)

200605 blog image Patchliner Webinar Playback

Marc will outline the development, optimization and validation of a range of voltage-gated Ca2+ channel assays on the Patchliner automated patch clamp platform that were subsequently used in an 8 year drug discovery collaboration between Metrion Biosciences and a german pharma company.

Speakers:
Dr. Marc Rogers (Chief Scientific Officer, Metrion Biosciences)
Dr. András Horváth (Application Scientist, Nanion Technologies)

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