Polycystin-2 Expression and Function in Adult Mouse Lacrimal Acinar Cells

Kaja, Simon; Hilgenberg, Jill D.; Rybalchenko, Volodymyr; Medina-Ortiz, Wanda E.; Gregg, Elaine V.; Koulen, Peter

doi:10.1167/iovs.10-7114

Cited by 17 publications

(19 citation statements)

References 34 publications

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“…In the absence of IP 3 and in the presence of 20 μM ruthenium red and 10 μM ryanodine to exclude activity from IP 3 receptors and ryanodine receptors, respectively, polycystin-2 single channel activity was recorded. As reported for kidney- and lacrimal gland-derived polycystin-2 channels (Koulen et al, 2002, Kaja et al, 2011b), polycystin-2 from RGCs also displayed a distinct voltage-dependent activity with increasing negative holding potentials leading to further activation that could be dose-dependently inhibited by increasing amounts of cytoplasmic Mg 2+ (Fig. 4A, B).…”

Section: Resultssupporting

confidence: 76%

“…1). Furthermore, specificity has previously shown by us and others (Tran et al, 2010, Kaja et al, 2011b). …”

Section: Resultssupporting

confidence: 65%

“…The specificity of the polyclonal antibody has previously been confirmed by us (Stokely et al, 2006), and we have recently described the expression and localization of polycystin-2 in the mouse lacrimal gland (Kaja et al, 2011b). The transcription factor Brn3a was used as a nuclear marker to positively identify RGCs in vertical cryostat sections (Quina et al, 2005, Nadal-Nicolas et al, 2009, Badea and Nathans, 2011).…”

Section: Discussionsupporting

confidence: 56%

“…ER membrane-enriched vesicles from immunopanned RGCs (obtained from retinae of 12 animals) were used in planar bilayer lipid membrane (BLM) electrophysiology as described previously (Koulen and Ehrlich, 2000, Koulen et al, 2001, Koulen et al, 2002, Westhoff et al, 2003, Koulen et al, 2005a, Koulen et al, 2005b, Hayrapetyan et al, 2008, Rybalchenko et al, 2008, Rybalchenko et al, 2009, Kaja et al, 2011b) using a Warner Instruments BLM workstation (Warner Instruments, LLC, Hamden, CT).…”

Section: Methodsmentioning

confidence: 99%

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Distribution and function of polycystin-2 in mouse retinal ganglion cells

et al. 2012

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The polycystin family of transient receptor potential (TRP) channels form Ca2+ regulated cation channels with distinct subcellullar localizations and functions. As part of heteromultimeric channels and multi-protein complexes, polycystins control intracellular Ca2+ signals and more generally the translation of extracellular signals and stimuli to intracellular responses. Polycystin-2 channels have been cloned from retina, but their distribution and function in retinal ganglion cells (RGCs) have not yet been established. In the present study, we determined cellular and subcellular localization as well as functional properties of polycystin-2 channels in RGCs. Polycystin-2 expression and distribution in RGCs was assessed by immunohistochemistry on vertical cryostat section of mouse retina as well as primary cultured mouse RGCs, using fluorescence microscopy. Biophysical and pharmacological properties of polycystin-2 channels isolated from primary cultured RGCs were determined using planar lipid bilayer electrophysiology. We detected polycystin-2 immunoreactivity both in the ganglion cell layer as well as in primary cultured RGCs. Subcellular analysis revealed strong cytosolic localization pattern of polycystin-2. Polycystin-2 channel current was Ca2+ activated, had a maximum slope conductance of 114 pS and could be blocked in a dose-dependent manner by increasing concentrations of Mg2+. The cytosolic localization of polycystin-2 in RGCs is in accordance with its function as intracellular Ca2+ release channel. We conclude that polycystin-2 forms functional channels in RGCs, of which biophysical and pharmacological properties are similar to polycystin-2 channels reported for other tissues and organisms. Our data suggest a potential role for polycystin-2 in RGC Ca2+ signaling.

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Section: Resultssupporting

confidence: 76%

“…1). Furthermore, specificity has previously shown by us and others (Tran et al, 2010, Kaja et al, 2011b). …”

Section: Resultssupporting

confidence: 65%

Section: Discussionsupporting

confidence: 56%

Section: Methodsmentioning

confidence: 99%

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Distribution and function of polycystin-2 in mouse retinal ganglion cells

et al. 2012

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“…This IP 3 subsequently causes Ca 2+ mobilization from internal stores (Berridge 2009). Release of Ca 2+ from intracellular stores can be mediated by three main groups of intracellular Ca 2+ channels in lacrimal gland: IP 3 receptors, ryanodine receptors, and polycystin family transient receptor potential channels (Yang et al 2002;Medina-Ortiz et al 2007;Kaja et al 2011). In addition, mechanisms that control Ca 2+ uptake into intracellular stores or extrusion of Ca 2+ into the extracellular space contribute critically to Ca 2+ homeostasis and the shape of Ca 2+ transients (Koulen and Thrower 2001;Berridge et al 2003).…”

Section: Evect Of Atp and Atp Analogs On [Ca 2+ ] I Dynamicsmentioning

confidence: 99%

P2Y purinoceptors induce changes in intracellular calcium in acinar cells of rat lacrimal glands

Kamada

Saino

Oikawa

et al. 2011

Histochem Cell Biol

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Adenosine 5'-triphosphate (ATP) is an extracellular signal that regulates various cellular functions. Cellular secretory activities are enhanced by ATP as well as by cholinergic and adrenergic stimuli. The present study aimed to determine which purinoceptors play a role in ATP-induced changes in the intracellular concentration of calcium ions ([Ca²⁺](i)) and in the fine structure of acinar cells of rat lacrimal glands. ATP induced exocytotic structures, vacuolation and an increase in [Ca²⁺](i) in acinar cells. The removal of extracellular Ca²⁺ or the use of Ca²⁺ channel blockers partially inhibited the ATP-induced [Ca²⁺](i) increase. U73122 (an antagonist of PLC) and heparin (an antagonist of IP₃ receptors) did not completely inhibit the ATP-induced [Ca²⁺](i) increase. P1 purinoceptor agonists did not induce any changes in [Ca²⁺](i), whereas suramin (an antagonist of P2 receptors) completely inhibited ATP-induced changes in [Ca²⁺](i). A P2Y receptor agonist, 2-MeSATP, induced a strong increase in [Ca²⁺](i), although UTP (a P2Y₂,₄,₆ receptor agonist) had no effect, and reactive blue 2 (a P2Y receptor antagonist) resulted in partial inhibition. The potency order of ATP analogs (2-MeSATP > ATP >>> UTP) suggested that P2Y₁ played a significant role in the cellular response to ATP. BzATP (a P2X₇ receptor agonist) induced a small increase in [Ca²⁺](i), but α,β-meATP (a P2X₁,₃ receptor agonist) had no effect. RT-PCR indicated that P2X₂,₃,₄,₅,₆,₇ and P2Y₁,₂,₄,₁₂,₁₄ are expressed in acinar cells. In conclusion, the response of acinar cells to ATP is mediated by P2Y (especially P2Y₁) as well as by P2X purinoceptors.

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