functional discrimination
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Lab on a Chip ◽  
2022 ◽  
Author(s):  
Bingyu B. Li ◽  
Erica Y. Scott ◽  
Ninni E. Olafsen ◽  
Jason Matthews ◽  
Aaron R. Wheeler

We studied the effect of AHR expression on metastasis using cell invasion in digital microfluidic microgel systems (CIMMS), which provided a unique combination of functional discrimination with transcriptome profiling of sub-populations of cells.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Mathilde Mathieu ◽  
Nathalie Névo ◽  
Mabel Jouve ◽  
José Ignacio Valenzuela ◽  
Mathieu Maurin ◽  
...  

AbstractDespite their roles in intercellular communications, the different populations of extracellular vesicles (EVs) and their secretion mechanisms are not fully characterized: how and to what extent EVs form as intraluminal vesicles of endocytic compartments (exosomes), or at the plasma membrane (PM) (ectosomes) remains unclear. Here we follow intracellular trafficking of the EV markers CD9 and CD63 from the endoplasmic reticulum to their residency compartment, respectively PM and late endosomes. We observe transient co-localization at both places, before they finally segregate. CD9 and a mutant CD63 stabilized at the PM are more abundantly released in EVs than CD63. Thus, in HeLa cells, ectosomes are more prominent than exosomes. By comparative proteomic analysis and differential response to neutralization of endosomal pH, we identify a few surface proteins likely specific of either exosomes (LAMP1) or ectosomes (BSG, SLC3A2). Our work sets the path for molecular and functional discrimination of exosomes and small ectosomes in any cell type.


2020 ◽  
Author(s):  
Mathilde Mathieu ◽  
Nathalie Névo ◽  
Mabel Jouve ◽  
José Ignacio Valenzuela ◽  
Mathieu Maurin ◽  
...  

ABSTRACTDespite their important and multiple roles in intercellular communications, the different populations of extracellular vesicles (EVs) and their secretion mechanisms are not fully characterized yet. In particular, how and to what extent EVs form either as intraluminal vesicles of endocytic compartments (exosomes), or at the plasma membrane (ectosomes) remains unclear. We followed in HeLa cells the intracellular trafficking of the EV markers CD9 and CD63 from the endoplasmic reticulum to their residency compartment and identified transient co-localization both at the plasma membrane (PM) and in endosomes, before they finally segregate. CD9 was more abundantly released in EVs than CD63. However, when forcing expression of CD63 at the PM, by mutating its lysosome-addressing motive, its secretion in EVs was increased. Thus, in HeLa cells, small ectosomes are more prominently released than exosomes. By comparative proteomic analysis, we identified a few surface proteins likely specific of either exosomes (e.g. LAMP1) or ectosomes (e.g. BSG, SLC3A2), based on their known intracellular location in lysosomes or the PM, and on the different effects on their release of Bafilomycin A1, a drug that neutralizes endosomal pH. Our work sets the path for molecular and functional discrimination of exosomes and small ectosomes in any cell type.


2020 ◽  
Vol 295 (49) ◽  
pp. 16614-16629
Author(s):  
Shiran Barber-Zucker ◽  
Jenny Hall ◽  
Afonso Froes ◽  
Sofiya Kolusheva ◽  
Fraser MacMillan ◽  
...  

Cation diffusion facilitator (CDF) proteins are a conserved family of divalent transition metal cation transporters. CDF proteins are usually composed of two domains: the transmembrane domain, in which the metal cations are transported through, and a regulatory cytoplasmic C-terminal domain (CTD). Each CDF protein transports either one specific metal or multiple metals from the cytoplasm, and it is not known whether the CTD takes an active regulatory role in metal recognition and discrimination during cation transport. Here, the model CDF protein MamM, an iron transporter from magnetotactic bacteria, was used to probe the role of the CTD in metal recognition and selectivity. Using a combination of biophysical and structural approaches, the binding of different metals to MamM CTD was characterized. Results reveal that different metals bind distinctively to MamM CTD in terms of their binding sites, thermodynamics, and binding-dependent conformations, both in crystal form and in solution, which suggests a varying level of functional discrimination between CDF domains. Furthermore, these results provide the first direct evidence that CDF CTDs play a role in metal selectivity. We demonstrate that MamM's CTD can discriminate against Mn2+, supporting its postulated role in preventing magnetite formation poisoning in magnetotactic bacteria via Mn2+ incorporation.


2019 ◽  
Vol 44 (5) ◽  
pp. 670-697
Author(s):  
Sigmund Eldevik ◽  
Hege Aarlie ◽  
Kristine B. Titlestad ◽  
Ellie Kazemi ◽  
Greg Elsky

The success of behavior-analytic procedures to teach language to individuals with autism spectrum disorder (ASD) has been well established in the literature. Nevertheless, some individuals may not learn any receptive or expressive language following extensive teaching efforts. We examined the effects of two reinforcement contingencies, functional and arbitrary, on increasing the level of auditory–visual conditional discriminations in children with ASD with a history of having difficulty learning discriminations. We evaluated the effects of the reinforcement contingencies by comparing the number of trials needed to establish discriminations in an adapted alternating treatment design. We found that five out of eight participants showed more rapid acquisition and demonstrated discrimination between more items in the functional reinforcement condition. The remaining three participants did not exhibit any discrimination in either condition within the allotted 500 trials/20 days. These findings suggest that using functional reinforcement procedures may be a helpful alternative for individuals who do not learn discriminations through traditional procedures.


2015 ◽  
Vol 8 (405) ◽  
pp. ra123-ra123 ◽  
Author(s):  
Ikuo Masuho ◽  
Olga Ostrovskaya ◽  
Grant M. Kramer ◽  
Christopher D. Jones ◽  
Keqiang Xie ◽  
...  

2013 ◽  
Vol 333 ◽  
pp. e699
Author(s):  
T. Fedele ◽  
H.-J. Scheer ◽  
M. Burghoff ◽  
G. Waterstraat ◽  
V. Nikulin ◽  
...  

2013 ◽  
Vol 44 (01) ◽  
Author(s):  
T Fedele ◽  
HJ Scheer ◽  
M Burghoff ◽  
G Waterstraat ◽  
V Nikulin ◽  
...  

PLoS ONE ◽  
2012 ◽  
Vol 7 (6) ◽  
pp. e39831 ◽  
Author(s):  
Daniel Fiole ◽  
Cédric Touvrey ◽  
Anne Quesnel-Hellmann ◽  
Julien Douady ◽  
Jean-Nicolas Tournier

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