Assessing the Secretory Capacity of Pancreatic Acinar Cells

Geron, Erez; Schejter, Eyal D.; Shilo, Ben‐Zion

doi:10.3791/51799

Cited by 5 publications

(4 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mouse pancreatic acinar cell clusters were isolated as previously described [ 27 , 28 ], with some adaptations. Briefly, the collected pancreas was minced into 1–3 mm 3 pieces and placed in ice-cold HBSS.…”

Section: Methodsmentioning

confidence: 99%

Thiopurines impair the apical plasma membrane expression of CFTR in pancreatic ductal cells via RAC1 inhibition

Tél

Papp

Varga

et al. 2023

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

Background and aims Thiopurine-induced acute pancreatitis (TIP) is one of the most common adverse events among inflammatory bowel disease patients treated with azathioprine (AZA), representing a significant clinical burden. Previous studies focused on immune-mediated processes, however, the exact pathomechanism of TIP is essentially unclear. Methods To model TIP in vivo, we triggered cerulein-induced experimental pancreatitis in mice receiving a daily oral dose of 1.5 mg/kg AZA. Also, freshly isolated mouse pancreatic cells were exposed to AZA ex vivo, and acinar cell viability, ductal and acinar Ca2+ signaling, ductal Cl– and HCO3– secretion, as well as cystic fibrosis transmembrane conductance regulator (CFTR) expression were assessed using microscopy techniques. Ras-related C3 botulinum toxin substrate (RAC1) activity was measured with a G-LISA assay. Super-resolution microscopy was used to determine protein colocalization. Results We demonstrated that AZA treatment increases tissue damage in the early phase of cerulein-induced pancreatitis in vivo. Also, both per os and ex vivo AZA exposure impaired pancreatic fluid and ductal HCO3– and Cl– secretion, but did not affect acinar cells. Furthermore, ex vivo AZA exposure also inhibited RAC1 activity in ductal cells leading to decreased co-localization of CFTR and the anchor protein ezrin, resulting in impaired plasma membrane localization of CFTR. Conclusions AZA impaired the ductal HCO3– and Cl– secretion through the inhibition of RAC1 activity leading to diminished ezrin-CFTR interaction and disturbed apical plasma membrane expression of CFTR. We report a novel direct toxic effect of AZA on pancreatic ductal cells and suggest that the restoration of ductal function might help to prevent TIP in the future.

show abstract

Section: Methodsmentioning

confidence: 99%

Thiopurines impair the apical plasma membrane expression of CFTR in pancreatic ductal cells via RAC1 inhibition

Tél

Papp

Varga

et al. 2023

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

show abstract

“…Previous protocols for exocrine cell isolation and culturing ( Gout et al., 2013 ) or isolation and adenoviral infections ( Geron et al., 2014 ) were published. Here, we optimized an ex vivo protocol for isolation, adenoviral infection and culturing of primary exocrine cells for up to 8 days.…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Protocol for Studying Reprogramming of Mouse Pancreatic Acinar Cells to β-like Cells

Elhanani

Walker

2020

STAR Protocols

View full text Add to dashboard Cite

Summary The potential of reprogrammed β cells derived from pancreatic exocrine cells to treat diabetes has been demonstrated in animal models. However, the precise mechanisms and regulators involved in this process are not clear. Here, we describe a method that allows mechanistic studies of this process in primary exocrine cultures using adenoviral expression vectors. This rapid 5-day protocol, provides the researcher with a highly controlled experimental system in which the effects of different compounds or genetic manipulations can be studied. For complete details on the use and execution of this protocol, please refer to Elhanani et al. (2020) .

show abstract

“…The latter accounts for 96-99% of the total pancreatic volume [12], and a schematic representation of the pancreatic anatomy is shown in Figure 1. The functional unit of the exocrine pancreas consist of an acinus and its corresponding draining ductile [19]. Pyramid-shaped acinar cells synthesize, store, and secrete digestive enzymes that are concentrated in a bicarbonate-rich fluid produced by ductal cells [20][21][22].…”

Section: The Pancreas 21 Anatomy and Physiology Of The Pancreasmentioning

confidence: 99%

In Vitro Disease Models of the Endocrine Pancreas

et al. 2021

View full text Add to dashboard Cite

The ethical constraints and shortcomings of animal models, combined with the demand to study disease pathogenesis under controlled conditions, are giving rise to a new field at the interface of tissue engineering and pathophysiology, which focuses on the development of in vitro models of disease. In vitro models are defined as synthetic experimental systems that contain living human cells and mimic tissue- and organ-level physiology in vitro by taking advantage of recent advances in tissue engineering and microfabrication. This review provides an overview of in vitro models and focuses specifically on in vitro disease models of the endocrine pancreas and diabetes. First, we briefly review the anatomy, physiology, and pathophysiology of the human pancreas, with an emphasis on islets of Langerhans and beta cell dysfunction. We then discuss different types of in vitro models and fundamental elements that should be considered when developing an in vitro disease model. Finally, we review the current state and breakthroughs in the field of pancreatic in vitro models and conclude with some challenges that need to be addressed in the future development of in vitro models.

show abstract

Assessing the Secretory Capacity of Pancreatic Acinar Cells

Cited by 5 publications

References 13 publications

Thiopurines impair the apical plasma membrane expression of CFTR in pancreatic ductal cells via RAC1 inhibition

Thiopurines impair the apical plasma membrane expression of CFTR in pancreatic ductal cells via RAC1 inhibition

Protocol for Studying Reprogramming of Mouse Pancreatic Acinar Cells to β-like Cells

In Vitro Disease Models of the Endocrine Pancreas

Contact Info

Product

Resources

About