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Biology subjects

Chalmers, S. B.

Publications and source records attributed to Chalmers, S. B..

3 recordsLinked to original sources

Macrophage calcium signaling dynamics revealed using genetically-encoded sensors

Macrophages (M{Phi}s) inhabit all mammalian organs, adapting to and influencing the tissues in which they reside. These versatile cells rapidly respond to tissue-specific signals, clean up debris and serve as the first line of defense against pathogens. Their ability to act quickly relates to their abundance and specific arrangement in tissues, as well as the systems they possess for detecting, processing and integrating emergent signals. Ca2+ is a fast second messenger that has been linked to many of the core functions of M{Phi}s. However, spatiotemporal features of physiologically-relevant Ca2+ signals in M{Phi}s remain largely uncharacterized. Using mouse models expressing genetic biosensors and fluorescently tagged channels, we visualize M{Phi} Ca2+ signal dynamics and heterogeneity, uncover a remarkable degree of mechanosensitivity in these cells, and characterize the physiological consequences of genetic ablation of Piezo1 channels via analysis of knockout models across their lifetime. Our in-depth investigation of M{Phi} Ca2+ signaling dynamics has broad relevance for the field of M{Phi} biology and the tissues that these cells support.

immunology↗

Simultaneous single cell imaging of calcium signal dynamics in breast cancer and neural cells reveals communication in a model of brain metastasis

The brain provides a unique metastatic microenvironment for breast cancer cells, where calcium signaling dynamics play critical roles in both cancer cell behavior and normal brain function. Calcium signal-mediated communication between breast cancer and neural cells has not yet been demonstrated through selective activation of breast cancer cells at the single cell level. To address this, we combined neural matrices of differentiated human neural progenitor cells with breast cancer cells expressing spectrally distinct genetically encoded calcium indicators. Specific activation of breast cancer cells increased calcium signaling activity in neural matrices with distinct temporal and spatial characteristics. Neural matrices also remodeled the expression of calcium-sensitive transcription factor SOX2 in a manner dependent on proximity to breast cancer cells. This work is the first simultaneous single-cell assessment of calcium signaling dynamics between breast cancer cells and neural cells modelling interactions within the brain metastatic niche.

cancer biology↗

Methods for imaging intracellular calcium signals in the mouse mammary epithelium in 2- and 3-dimensions

The mammary gland has a central role in optimal mammalian development and survival. Contractions of smooth muscle-like basal (or myoepithelial) cells in the functionally mature mammary gland in response to oxytocin are essential for milk ejection and are tightly regulated by intracellular calcium (Ca2+). Using mice expressing a genetically encoded Ca2+ indicator (GCaMP6f), we present in this chapter a method to visualise at high spatiotemporal resolution changes in intracellular Ca2+ in mammary epithelial cells, both in vitro (2D) and ex vivo (3D). The procedure to optimally prepare mammary tissue and primary cells is presented in detail.

physiology↗