Keratinocyte Calcium Response
Keratinocyte Calcium Response - Related Videos
05:36
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This video describes the isolation of keratinocytes—the most prominent cells of human skin's epidermal layer, by employing a combination of enzymatic digestion and mechanical disruption methods. This method helps in the preferential growth of keratinocytes over other skin cell types using a selective growth medium.
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Epidermal keratinocytes form a functional skin barrier and are positioned at the front line of host defense against external environmental insults. Here we describe methods for isolation and primary culture of epidermal keratinocytes from neonatal and adult mouse skin, and induction of terminal differentiation and UVB-triggered inflammatory response from keratinocytes.
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This video describes a detailed protocol for producing genetically modified primary human keratinocytes recombinant retrovirus. This method can be used to manipulate gene expression in keratinocytes.
06:44
Keratinocyte Colony Formation Assay: An In Vitro Assay to Assess the Ability of Keratinocytes to Grow into Colonies
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In this video, we perform a clonogenic assay of keratinocytes harvested from adult mice to determine the ability of individual cells to proliferate and form colonies. This keratinocyte harvesting technique is useful for studying other downstream aspects of cutaneous chemical carcinogenesis and tumor promotion.
03:43
An Imaging Technique to Study the Calcium Response in Bacteria-Infected Cells
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This video demonstrates a technique for detecting local and global calcium ion response upon Shigella infection of human epithelial cells. Shigella invades host cells and releases invasion effectors, impacting intracellular calcium levels. The amplitude and frequency of the local and global cytosolic calcium are studied using a fluorescent indicator.
FAQs
Stimulus-sensitive membrane channels and receptors determine how keratinocytes convert chemical, mechanical, thermal, or inflammatory inputs into calcium signals. They can promote calcium entry across the plasma membrane or regulate release from intracellular stores. The resulting signal may be transient or sustained, allowing researchers to distinguish rapid cellular reactions from responses that persist during ongoing stimulation.
Signal duration provides information about how keratinocytes respond over time. A transient increase can indicate a brief reaction to stimulation, whereas a sustained elevation may reflect continued signaling during persistent chemical, mechanical, thermal, or inflammatory conditions. Comparing these patterns helps investigators characterize cellular responsiveness and relate epidermal activity to processes such as inflammation, pain, itch, or repair.
Keratinocyte calcium signals do not depend only on calcium entering through the cell membrane. Stimulus-sensitive receptors and channels can also regulate calcium release from intracellular stores, adding another source of signal. Considering both routes is important when interpreting measurements, because similar changes in intracellular calcium may arise from membrane entry, store release, or combined activity.
Researchers can measure these responses with fluorescent calcium indicators that report changes in intracellular calcium concentration. Keratinocytes are exposed to a stimulus, and fluorescence is monitored to identify the timing and magnitude of the resulting signal. This approach allows investigators to compare transient and sustained responses across chemical, mechanical, thermal, or inflammatory conditions without treating all signals as equivalent.
Experimental studies may examine chemical, mechanical, thermal, or inflammatory stimuli because each represents a distinct way epidermal cells can be activated. Comparing these inputs helps determine whether calcium signaling varies with stimulus type and duration. The resulting measurements can support investigations of how skin-cell activity contributes to inflammation, wound repair, pain, itch, and sensory processing.
In a neurocutaneous context, keratinocyte calcium activity helps researchers examine communication between epidermal cells and nearby sensory nerve endings. Calcium signaling can accompany release of signaling molecules that influence those endings, providing a cellular link between skin events and peripheral nervous system function. This framework supports research on pain, itch, inflammation, and how sensory information begins in the skin.