Many G protein-coupled receptors (GPCRs) are internalized from the plasma membrane after agonist exposure. Previously, marked agonist-induced internalization of human alpha2A- and alpha2B-adrenergic receptors (AR) was observed in transfected neuronal rat pheochromocytoma (PC12) cells; alpha2A- and alpha2B-AR were internalized into partly distinct intracellular vesicles (Olli-Lähdesmäki et al., J. Neurosci. 19, 9281-9288, 1999). In this paper, the extent of alpha2-AR internalization was quantitated in human embryonic kidney (HEK-293) and PC12 cells by combined application of cell surface biotinylation and ELISA methods, which allow measurement of protein trafficking in intact, differentiated and undifferentiated cells. Significant subtype-specific (but not cell type-dependent) trafficking of human alpha2-AR was observed by quantitation and immunocytochemistry. Agonist-induced sequestration of alpha2B-AR was markedly reduced after blocking the formation of clathrin-coated vesicles by hyperosmotic sucrose pretreatment. The sequestration of alpha2A-AR was partly inhibited after sucrose pretreatment but could be further reduced after inhibiting the formation of both clathrin-coated and caveolin vesicles by combined pretreatment with hyperosmotic sucrose and filipin. Differences were also observed in the recycling of alpha2A- and alpha2B-AR. The extent of maximal agonist-induced sequestration in PC12 cells was not directly dependent on relative agonist efficacy.
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http://dx.doi.org/10.1078/0171-9335-00311 | DOI Listing |
Food Chem Toxicol
January 2025
Department of Environmental Health & Environment and Health Innovation Team, School of Public Health, Zhengzhou University, Zhengzhou, 450001, Henan, China. Electronic address:
Neurological dysfunction induced by fluoride is still one of major concern worldwide, yet the underlying mechanisms remain elusive. To explore whether fluoride disrupts lysosomal biosynthesis via the GSK3β signaling, leading to neurological damage, both in vivo rat models and in vitro PC12 cell models were conducted. Subsequent findings revealed reduced spatial learning and memory abilities, decreased hippocampal neurons, and disrupted neuronal arrangement in NaF-treated rats.
View Article and Find Full Text PDFTalanta
January 2025
Department of Pharmaceutical Analysis, Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Faculty of Pharmacy, Fujian Medical University, Fuzhou, 350122, China. Electronic address:
The rise of extracellular matrix (ECM)-supported three-dimensional (3D) cell culture systems which bridge the gap between in vitro culture and in vivo living tissue for pharmacological models has increased the need for simple and robust cell viability assays. This study presents the development of an effective biosensing assay for in situ monitoring of the catecholamine neurotransmitter exocytosis levels for cell viability assessment within complicated cell-encapsulated hydrogel milieu. Firstly, the biosensing assay demonstrated the distinction among four pheochromocytoma (PC12) cell lines with varying degrees of differentiation and the discrepancy in cellular neurosecretory capacity between two-dimensional (2D) monolayer and 3D agarose hydrogel culture conditions, accompanied by morphological distinctions.
View Article and Find Full Text PDFActa Biomater
January 2025
Central laboratory of Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China. Electronic address:
Peripheral nerve injury (PNI) as a common clinical issue that presents significant challenges for repair. Factors such as donor site morbidity from autologous transplantation, slow recovery of long-distance nerve damage, and deficiencies in local cytokines and extracellular matrix contribute to the complexity of effective PNI treatment. It is extremely urgent to develop functional nerve guidance conduits (NGCs) as substitutes for nerve autografts.
View Article and Find Full Text PDFBackground: Fibroblast growth factor 21 (FGF21) and Methyltransferase-like 14 (METTL14) have been identified to be involved in spinal cord injury (SCI). However, whether FGF21 functioned in SCI via METTL14-induced N6-methyladenosine (m6A) modification remains unclear.
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Int J Biol Macromol
January 2025
State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, PR China; State Key Laboratory of Food Nutrition and Safety, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, PR China; College of Food Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, PR China. Electronic address:
Morchella esculenta (L.) Pers. is considered a precious edible and medicinal fungus due to its strict growth environment requirements, difficult to cultivate, resulted in expensive in the market.
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