Interdependent Roles Research Articles

Mucosal Mast Cell-Specific Gene Expression Is Promoted by Interdependent Action of Notch and TGF-β Signaling.

Rodent mast cells are classified into two major subsets, mucosal mast cells (MMCs) and connective tissue mast cells. MMCs arise from mast cell progenitors that are mobilized from the bone marrow to mucosal tissues in response to allergic inflammation or helminth infection. TGF-β is known as an inducer of MMC differentiation in mucosal tissues, but we have previously found that Notch receptor-mediated signaling also leads to the differentiation. Here, we examined the relationship between Notch and TGF-β signaling in MMC differentiation using mouse bone marrow-derived mast cells (BMMCs). We found that the coexistence of Notch and TGF-β signaling markedly upregulates the expression of MMC markers, mouse mast cell protease (mMCP)-1, mMCP-2, and αE integrin/CD103, more than Notch or TGF-β signaling alone, and that their signals act interdependently to induce these marker expressions. Notch and TGF-β-mediated transcription of MMC marker genes were both dependent on the TGF-β signaling transducer SMAD4. In addition, we also found that Notch signaling markedly upregulated mMCP-1 and mMCP-2 expression levels through epigenetic deregulation of the promoter regions of these genes, but did not affect the promoter of the CD103-encoding gene. Moreover, forced expression of the constitutively active Notch2 intracellular domain in BMMCs showed that Notch signaling promotes the nuclear localization of SMADs 3 and 4 and causes SMAD4-dependent gene transcription. These findings indicate that Notch and TGF-β signaling play interdependent roles in inducing the differentiation and maturation of MMCs. These roles may contribute to the rapid expansion of the number of MMCs during allergic mucosal inflammation.

The interdependent roles of the psychosocial predictors of human papillomavirus vaccination among Christian parents of unvaccinated adolescents

ABSTRACT Despite the availability of the human papillomavirus (HPV) vaccine, uptake has been sub-optimal among certain religious groups. Psychosocial factors (threat appraisal, coping appraisal, and attitudes) have been identified as independent determinants of HPV vaccination. However, their interdependent effects have not been tested. We examined the interdependency of these psychosocial factors in predicting HPV vaccination intention among Christian parents of unvaccinated adolescents (using a theory-driven conceptual model). A cross-sectional study of 342 participants showed that perceived self-efficacy (β = 0.2, 0.11–0.29, p = <0.0001) and perceived response efficacy of HPV vaccine (β = 0.65, 0.53–0.77. p < .0001) were positively associated with vaccination intention. Our mediation analysis (using the Preachers and Hayes’ approach) shows that attitudes toward HPV vaccination mediated 59% of the relationship between perceived self-efficacy to vaccinate child and HPV vaccination intention; and 61% of the relationship between perceived response efficacy of HPV vaccine and HPV vaccination intention. Attitudes may be the psychosocial factor that drives the effects of coping appraisal. Therefore, designing an attitude-based intervention to address religious barrier beliefs among Christian parents may nullify the impact of low self-efficacy and response efficacy on HPV vaccination.

Membrane dynamics of ATG4B and LC3 in autophagosome formation

The biogenesis of autophagosomes provides the basis for macroautophagy to capture and degrade intracellular cargoes. Binding of the autophagy-related protein ATG8/LC3 to autophagic membranes is essential to autophagosome formation, which involves the specific and dynamic processing of ATG8/LC3 by cysteine protease ATG4. However, to date, the mechanism whereby ATG4 is recruited to the membranes, the interaction of ATG4 and ATG8/LC3 on the membranes, and its role in the growth of phagophore are not completely understood. Here, we used fluorescence recovery after photobleaching to monitor the turnover of GFP-tagged ATG4B and LC3B in living animal cells. The data show that ATG4B localizes to early autophagic membranes in an LC3B-dependent manner. During autophagy, ATG4B and LC3B undergo rapid cytosol/isolation membrane exchange but not at the cytosol/completed autophagosome. In addition, ATG4B activity controls the efficiency of autophagosome formation by impacting the membrane binding/dissociation of LC3B. These data suggest that ATG4 and LC3 play interdependent roles in the formation of autophagosomes.

INVESTIGATING THE ROLE OF COGNITIVE VARIABLES IN SECOND LANGUAGE LEARNERS’ LISTENING COMPREHENSION: APTITUDE AND METACOGNITIVE AWARENESS

ABSTRACT The influence of individual difference variables on second language (L2) listening comprehension has been an area of research that has been gaining renewed attention in recent years. The current study investigated the effects of two cognitive variables, aptitude and metacognitive awareness, on 107 native-Korean, English as a foreign language learners’ listening comprehension. Moreover, the study examined whether metacognitive awareness mediated the relationship between aptitude and L2 listening comprehension. Both aptitude and metacognitive awareness were found to have statistically significant positive correlations (r = .58 and .37, respectively) with L2 listening comprehension, and they were also significant predictors of L2 listening, collectively explaining 41% of the variance in L2 listening performance. Furthermore, metacognitive awareness mediated the relationship between aptitude and L2 listening comprehension. These findings suggest that cognitive learner variables play a significant and interdependent role in L2 listening comprehension.

Spatiotemporal blood vessel specification at the osteogenesis and angiogenesis interface of biomimetic nanofiber-enabled bone tissue engineering

While extensive research has demonstrated an interdependent role of osteogenesis and angiogenesis in bone tissue engineering, little is known about how functional blood vessel networks are organized to initiate and facilitate bone tissue regeneration. Building upon the success of a biomimetic composite nanofibrous construct capable of supporting donor progenitor cell-dependent regeneration, we examined the angiogenic response and spatiotemporal blood vessel specification at the osteogenesis and angiogenesis interface of cranial bone defect repair utilizing high resolution multiphoton laser scanning microscopy (MPLSM) in conjunction with intravital imaging. We demonstrate here that the regenerative vasculature can be specified as arterial and venous capillary vessels based upon endothelial surface markers of CD31 and Endomucin (EMCN), with CD31+EMCN− vessels exhibiting higher flowrate and higher oxygen tension (pO2) than CD31+EMCN+ vessels. The donor osteoblast clusters are uniquely coupled to the sprouting CD31+EMCN+ vessels connecting to CD31+EMCN− vessels. Further analyses reveal differential vascular response and vessel type distribution in healing and non-healing defects, associated with changes of gene sets that control sprouting and morphogenesis of blood vessels. Collectively, our study highlights the key role of spatiotemporal vessel type distribution in bone tissue engineering, offering new insights for devising more effective vascularization strategies for bone tissue engineering.

CRISPR/Cas9‐mediated knockout of G proteins and β‐arrestins defines their specific but interdependent roles in dopamine D1 receptor signaling

The dopamine D1 receptor (D1R) is a G protein-coupled receptor which regulates key brain functions including voluntary movement, memory, attention, and reward. Selective agonist activation of the D1R may also provide novel neurotherapeutics for neurodegenerative and neuropsychiatric disorders. The D1R canonically activates the heterotrimeric G proteins Gαs, and possibly Gαolf, which activate adenylyl cyclase (AC) to increase cAMP/PKA signaling. The D1R also engages β-arrestin proteins which may transduce β-arrestin dependent signaling by scaffolding and activating kinases. However, the distinct roles of Gαs, Gαolf versus β-arrestins 1 and 2 for D1R signaling remains largely undefined. Here we utilize CRISPR/Cas9 engineered HEK293 cells in which Gαs, Gαolf, β-arrestin1 and β-arrestin2 are stably knocked out (KO) to delineate D1R pharmacology and signaling mechanisms. Western blots confirmed a complete loss of Gαs, Gαolf, β-arrestin1 or β-arrestin2 proteins in KO cells versus parental wildtype cells. Dose responses with the D1R agonist SKF-81297 or β2AR agonist isoproterenol induced robust cAMP production in wildtype cells that was eliminated in Gαs/olf KO cells. Re-expression of Gαs or Gαolf in KO cells rescued D1R-mediated cAMP signaling similar to wildtype cells. Interestingly, the D1R had significantly higher cAMP signaling when coupling via Gαs as compared to Gαolf, while the β2AR only coupled with Gαs to stimulate cAMP signaling. Western blotting indicated Gαs/olf KO completely blocked agonist induced CREB phosphorylation by both D1R and β2AR agonists, indicating a strict dependence on Gαs/olf and cAMP/PKA signaling to activate CREB. In major contrast, β-arrestin1/2 KO significantly increased the intensity and duration of cAMP signaling and CREB phosphorylation by the β2AR. β-arrestin1/2 KO also entirely blocked agonist-induced D1R internalization and impaired D1R desensitization while Gαs/olf KO had no effect on D1R internalization. Key controls of re-expressing Gαs, Gαolf, β-arrestin1 or β-arrestin2 in KO cells completely restored signaling or internalization similar to wildtype cells, indicating CRISPR/Cas9 targeting did not result in non-specific or compensatory effects in the pathways under study. Notably, β-arrestin1/2 KO also potentiated and prolonged D1R or β2AR-induced ERK1/2 phosphorylation. D1R-induced ERK1/2 phosphorylation in β-arrestin1/2 KO cells was eliminated by the SRC inhibitor SU6656 or the MEK inhibitor U0126, suggesting that D1R signaling via a Gαs/SRC/MEK pathway is essential for ERK1/2 activation. Taken together, our studies indicate that Gαs is the dominant transducer for D1R cAMP, CREB and ERK1/2 signaling and that β-arrestins provide distinctly interdependent roles with G proteins in modulating the duration and intensity of D1R and β2AR signaling cascades. Using CRISPR/Cas9 KO model cells, future studies will examine the distinct contributions of β-arrestins 1 and 2 and Gαs/olf for D1R pharmacology, which will be useful in the development of novel D1R biased agonists.

Diet and lung disease-Are fruits and vegetables the ideal whole-food intervention?

Diet and lung disease-Are fruits and vegetables the ideal whole-food intervention?

Protein Disulphide Isomerase and NADPH Oxidase 1 Cooperate to Control Platelet Function and Are Associated with Cardiometabolic Disease Risk Factors.

Background: Protein disulphide isomerase (PDI) and NADPH oxidase 1 (Nox-1) regulate platelet function and reactive oxygen species (ROS) generation, suggesting potentially interdependent roles. Increased platelet reactivity and ROS production have been correlated with cardiometabolic disease risk factors. Objectives: To establish whether PDI and Nox-1 cooperate to control platelet function. Methods: Immunofluorescence microscopy was utilised to determine expression and localisation of PDI and Nox-1. Platelet aggregation, fibrinogen binding, P-selectin exposure, spreading and calcium mobilization were measured as markers of platelet function. A cross-sectional population study (n = 136) was conducted to assess the relationship between platelet PDI and Nox-1 levels and cardiometabolic risk factors. Results: PDI and Nox-1 co-localized upon activation induced by the collagen receptor GPVI. Co-inhibition of PDI and Nox-1 led to additive inhibition of GPVI-mediated platelet aggregation, activation and calcium flux. This was confirmed in murine Nox-1−/− platelets treated with PDI inhibitor bepristat, without affecting bleeding. PDI and Nox-1 together contributed to GPVI signalling that involved the phosphorylation of p38 MAPK, p47phox, PKC and Akt. Platelet PDI and Nox-1 levels were upregulated in obesity, with platelet Nox-1 also elevated in hypertensive individuals. Conclusions: We show that PDI and Nox-1 cooperate to control platelet function and are associated with cardiometabolic risk factors.

What mediates the racial/ethnic disparity in psychosocial stress among breast cancer patients?

BackgroundPrior studies have observed greater levels of psychosocial stress (PSS) among non-Hispanic (nH) African American and Hispanic women when compared to nH White patients after a breast cancer diagnosis. We aimed to determine the independent and interdependent roles of socioeconomic position (SEP) and unmet support in the racial disparity in PSS among breast cancer patients.MethodsParticipants were recruited from the Breast Cancer Care in Chicago study (n = 989). For all recently diagnosed breast cancer patients, aged 25–79, income, education, and tract-level disadvantage and affluence were summed to create a standardized socioeconomic position (SEP) score. Three measures of PSS related to loneliness, perceived stress, and psychological consequences of a breast cancer diagnosis were defined based on previously validated scales. Five domains of unmet social support needs (emotional, spiritual, informational, financial, and practical) were defined from interviews. We conducted path models in MPlus to estimate the extent to which PSS disparities were mediated by SEP and unmet social support needs.ResultsBlack and Hispanic patients reported greater PSS compared to white patients and greater unmet social support needs (p = 0.001 for all domains). Virtually all of the disparity in PSS could be explained by SEP. A substantial portion of the mediating influence of SEP was further transmitted by unmet financial and practical needs among Black patients and by unmet emotional needs for Hispanic patients.ConclusionsSEP appeared to be a root cause of the racial/ethnic disparities in PSS within our sample. Our findings further suggest that different interventions may be necessary to alleviate the burden of SEP for nH AA (i.e., more financial support) and Hispanic patients (i.e., more emotional support).

Discordant hepatic fatty acid oxidation and triglyceride hydrolysis leads to liver disease.

To extract energy from stored lipids, fatty acids must first be liberated from triglyceride before their β-oxidation in mitochondria in a coordinated and stepwise manner. To determine the independent and interdependent roles of hepatic triglyceride hydrolysis and fatty acid oxidation, mice were generated with a liver-specific defect in triglyceride hydrolysis (AtglL–/–), fatty acid oxidation (Cpt2L–/–), or both (double knockout). The loss of either gene resulted in the compensatory increase in the other, demonstrating their coordination. The loss of individual components of fatty acid catabolism (carnitine palmitoyl transferase 2 [Cpt2], adipose triglyceride lipase [Atgl], and Pparα) resulted in largely independent effects on hepatocyte morphology, intermediary metabolism, and gene expression in response to fasting. However, high-fat feeding revealed the interdependent role of Atgl and Cpt2, as the loss of only one of the genes resulted in steatosis (fatty liver) but the loss of both components resulted in significant steatohepatitis (inflammation and fibrosis). Lipolysis and β-oxidation are intimately linked within a continuous pathway, and disruption of their coordination leads to unique cellular and molecular phenotypes that ultimately result in liver disease.

Key Approaches to the Study of History of Altai in 18th Century in Modern Russian Historiography

The article is devoted to a comprehensive examination of theoretical approaches to the study of the history of Altai in the 18th century, developed by Russian specialists over the past three decades. The relevance of the study is associated with the emerging trend in domestic science towards a holistic understanding of the retrospective of individual regions, including through understanding the interdependent role of various social and administrative factors in the colonization process. Special attention is paid to the conceptual assessment of the nature of the initial development of Altai. The methodological grounds for applying the theories of military colonization, frontier, frontier modernization to the history of the region are outlined. The authors state the presence in the scientific literature of two non-identical approaches to the characterization of the Russian border in the south of Western Siberia. The authors demonstrate the obvious desire of historians to carry out comprehensive studies of regional and local levels of civil and mining management, which has emerged against the background of the preservation of the heuristic significance of microhistorical analysis of the role of individual fortified (fortresses, forts) or production facilities (factories). The diversity and conceptual heterogeneity of modern Russian historiography of the history of Altai in the 18th century is summarized. Some forecasts of the development of the historiographic situation are formulated.

Masculine and feminine: Interdependent gender roles in ‘fight club’

Masculine and feminine: Interdependent gender roles in ‘fight club’

The Intra-Action of Employees and Materiality in the Constitution of Work-Related Identity

Applying a sociomaterial view to the material and human agency in doing work, this study explores the interdependent role of material arrangements and people doing knowledge-work. Using a Nordic university organization as an example, the study specifically highlights how material-physical environment and its affordances, including spaces of work as well as “high technology” such as electronics and “low technology” such as furniture – gain agency and become significant in terms of work-related identity. We introduce three modes of work: cognitive-, embodied-, and emotional work, which describe cognitive, bodily, and emotional actions that are entangled in the expressions of knowledge workers’ work-related identities. Focusing on identity as an achievement in agentive “intra-action” between the human and the material, the article contributes to the research on the effects of material arrangements and technology in the activities that knowledge-work in a digitalized workplace in particular consists of. Keywords: work-related identity, sociomateriality, material agency

Impact of lean operations on the roles of finance functions and their application of lean

PurposeThis paper aims to study the relations between lean operations, lean principles in finance functions and the roles of finance functions.Design/methodology/approachThe paper uses structural equation modeling to analyze data from 408 different firms in the Danish production and services sectors. A dyadic approach is applied, as a sub-sample of 107 chief operating officers in the responding firms is used to investigate the construct validity, reliability and average deviation index of the instrument measuring the roles of finance functions.FindingsThe paper finds that lean-operation firms emphasize four different yet interdependent roles of finance functions. The paper also finds that lean operation leads to firms’ finance functions adopting lean principles.Research limitations/implicationsThis paper characterizes lean-operation firms as contextually ambidextrous to predict relations between lean operation and roles of finance functions. The paper expands prior case study findings on the roles of finance functions in lean-operation firms, and the findings of the paper underline that finance functions continue to play an important role in these firms.Practical implicationsDecision-makers in lean-operation firms should not be hesitant with respect to integrating finance function workers into the lean operation. Furthermore, decision-makers should understand that a balanced emphasis of the roles of finance functions is necessary to avoid overemphasizing exploitation at the expense of exploration, or vice versa.Originality/valueTo the best of the authors’ knowledge, this is the first paper to provide large-scale evidence of the roles of finance functions in lean-operation firms and to show that lean principles diffuse to finance functions. Furthermore, the paper introduces a new instrument for measuring finance function roles, based on the competing values framework.

EUROPEAN UNION’S COMMITMENT TO HUMAN RIGHTS: LEGAL FOUNDATION AND OTHER POLICIES TO PROMOTE AND SAFEGUARD HUMAN RIGHTS WITHIN EUROPEAN UNION

The European Union (EU) maintains that a legitimate interest of the international community is the promotion and protection of human rights around the world. In fostering human rights, democratization and development, the European Union is bound by its founding Treaties and other policies. The policies of the European Union in promotion of human rights are focused on globally agreed structures and norms. Throughout its development, the EU has progressively acceded to the concept of human rights, and promoted their fundamental, indivisible and interdependent role. European Union believes that human rights are indivisible and holds resolute tone against the challenges of universal human rights.

Virtual Reality Simulation in Interprofessional Round Training for Health Care Students: A Qualitative Evaluation Study

BackgroundInterprofessional rounds are increasingly applied in providing patient-centered team care delivery. Students from six health care courses in medicine, nursing, pharmacy, physiotherapy, occupational therapy, and social work formed interprofessional teams to undertake an interprofessional education (IPE) on virtual reality simulation–based interprofessional rounds. This study aimed to evaluate health care students’ perspectives on the transferability of the IPE virtual reality simulation learning to clinical practice. MethodThree focus group discussions were conducted. ResultsThree themes emerged: “gaining insights into mutual roles,” whereby students better understood the different interprofessional roles of other team members, “seeing the patient as a whole” to foster patient-centered care, and “gaps in real-world application” due to system and structure constraints. ConclusionsEarly exposure to team care through IPE virtual reality simulation can foster understandings of the interdependent roles of health care professionals toward patient-centered care. For greater clinical impact, a further recommendation is to supplement with workplace-based team training to contextualize learning with practice settings.

Spatiotemporal Modulation of Flavonoid Metabolism in Blueberries.

Blueberries are distinguished by their purple-blue fruit color, which develops during ripening and is derived from a characteristic composition of flavonoid-derived anthocyanin pigments. The production of anthocyanins is confined to fruit skin, leaving the colorless fruit flesh devoid of these compounds. By linking accumulation patterns of phenolic metabolites with gene transcription in Northern Highbush (Vaccinium corymbosum) and Rabbiteye (Vaccinium virgatum) blueberry, we investigated factors limiting anthocyanin production in berry flesh. We find that flavonoid production was generally lower in fruit flesh compared with skin and concentrations further declined during maturation. A common set of structural genes was identified across both species, indicating that tissue-specific flavonoid biosynthesis was dependent on co-expression of multiple pathway genes and limited by the phenylpropanoid pathway in combination with CHS, F3H, and ANS as potential pathway bottlenecks. While metabolite concentrations were comparable between the blueberry genotypes when fully ripe, the anthocyanin composition was distinct and depended on the degree of hydroxylation/methoxylation of the anthocyanidin moiety in combination with genotype-specific glycosylation patterns. Co-correlation analysis of phenolic metabolites with pathway structural genes revealed characteristic isoforms of O-methyltransferases and UDP-glucose:flavonoid-3-O-glycosyltransferase that were likely to modulate anthocyanin composition. Finally, we identified candidate transcriptional regulators that were co-expressed with structural genes, including the activators MYBA, MYBPA1, and bHLH2 together with the repressor MYBC2, which suggested an interdependent role in anthocyanin regulation.

Cognition and communication: situational awareness and tie preservation in disrupted task environments

AbstractIndividuals filling specialized, interdependent organizational roles achieve coordinated task execution through effective communication channels. Such channels enable regular access to information, opportunities, and assistance that may enhance one’s understanding of the task environment. However, the time and effort devoted to maintaining those channels may detract from one’s duties by turning attention away from the task environment. Disrupted task environments increase information requirements, thus creating a dilemma in which individuals must sustain benefits offered by important communication channels and relieve burdens imposed by ineffective channels. Using separable temporal exponential random graph models (STERGMs), this paper examines the relationship between situational awareness (SA) and the propensity to sustain or dissolve preexisting communication channels during 10 disruptive events experienced sequentially by a large, multifaceted military organization during a 2-week training exercise. Results provide limited evidence that increased SA detracts from tie preservation; instead SA begins to predict tie preservation during the second week of the exercise. Patterns of organizational adaptation reveal that, over time, improvised coordinative roles increasingly fall upon those with elevated SA. These results suggest that over successive disruptions, the benefits of information provided by communication channels within interdependent, role-specialized organizations begin to outweigh the costs of sustaining those channels.

Acquiring environmental flows: ecological economics of policy development in western U.S.

The western U.S. faces trade-offs between providing water for urban, agricultural and industrial uses and allocating water to environmental needs. These competing uses have been the subject of many conflicts addressed by courts, legislatures and voluntary water trading arrangements. This paper draws upon Social Ecological Economics of Water (SEEW) and Punctuated Equilibrium Theory (PET) to evaluate the interdependent roles of voluntary transactions and mandates to provide environmental water. The article presents stages of maturation in water trading, and applies these to two evolving markets in the western U.S. Case studies of the Edwards Aquifer in Texas and the Colorado River Delta illustrate the interplay of voluntary water trading and compulsory reallocation. The cases highlight interdependencies among policy instruments and pathways of water policy innovation.The SEEW and PET frameworks shed light on persistent difficulties faced with developing markets to acquire water for environmental needs. The case studies demonstrate SEEW concepts of patrimony and power dynamics in considering why objections to voluntary water trading endure, even though water transactions are a valuable policy instrument. The case studies illustrate PET principles on the role of economic impetus in propelling policy innovation and on pivotal contributions by academics and NGOs.

CRISPR/Cas9‐mediated knockout of G proteins and β‐arrestins delineates their distinct but interdependent roles in dopamine D1 receptor signaling

The dopamine D1 receptor (D1R) is a G protein‐coupled neurotransmitter receptor which controls key brain functions including voluntary movement, memory, attention and reward. Selective agonist activation of the D1R may also provide novel neurotherapeutics for neurodegenerative and neuropsychiatric disorders. The D1R canonically activates the heterotrimeric G proteins Gαs, and possibly Gαolf, which activate adenylyl cyclase (AC) to increase cAMP/PKA signaling. The D1R also engages β‐arrestin proteins which may transduce β‐arrestin dependent signaling by scaffolding and activating kinases. However, the distinct roles of Gαs, Gαolf versus β‐arrestins 1 and 2 for D1R signaling remains largely undefined. Here we utilize CRISPR/Cas9 engineered HEK293 cells in which Gαs, Gαolf, β‐arrestin1 and β‐arrestin2 are stably knocked out (KO) to delineate D1R pharmacology and signaling mechanisms. Western blots confirmed a complete loss of Gαs, Gαolf, β‐arrestin1 or β‐arrestin2 proteins in KO cells versus parental wildtype cells. Dose responses with the D1R agonist SKF‐81297 or β2AR agonist isoproterenol induced robust cAMP production in wildtype cells that was eliminated in Gαs/olf KO cells. Re‐expression of Gαs or Gαolf in KO cells rescued D1R‐mediated cAMP signaling similar to wildtype cells. Notably, the D1R had significantly higher cAMP signaling when coupling via Gαs as compared to Gαolf, while the β2AR only coupled with Gαs to stimulate cAMP signaling. Western blotting indicated Gαs/olf KO completely blocked agonist‐induced CREB phosphorylation by both D1R and β2AR agonists, indicating a strict dependence on Gαs/olf and cAMP/PKA signaling to activate CREB. In major contrast, β‐arrestin1/2 KO significantly increased the intensity and duration of cAMP signaling and CREB phosphorylation by the β2AR. β‐arrestin1/2 KO also entirely blocked agonist‐induced D1R internalization and impaired D1R desensitization while Gαs/olf KO had no effect on D1R internalization. Key controls of re‐expressing Gαs, Gαolf, β‐arrestin1 or β‐arrestin2 in KO cells completely restored signaling or internalization similar to wildtype cells, indicating CRISPR/Cas9 targeting did not result in non‐specific or compensatory effects in the pathways under study. Notably, β‐arrestin1/2 KO also potentiated and prolonged D1R or β2AR‐induced ERK1/2 phosphorylation. D1R‐induced ERK1/2 phosphorylation in β‐arrestin1/2 KO cells was eliminated by the SRC inhibitor SU6656 or the MEK inhibitor U0126, suggesting that D1R signaling via a Gαs/SRC/MEK pathway is essential for ERK1/2 activation. Taken together, our studies indicate that Gαs is the dominant transducer for D1R cAMP, CREB and ERK1/2 signaling and that β‐arrestins provide distinctly interdependent roles with G proteins in modulating the duration and intensity of D1R and β2AR signaling cascades. Using CRISPR/Cas9 KO model cells, future studies will examine the distinct contributions of β‐arrestins and Gαs/olf for D1R pharmacology, which will be useful in defining and validating novel D1R biased agonists.Support or Funding InformationNIH DA047643 (JAA); Rising STAR award from the University of Texas System (JAA); Institutional funding from UTMB (JAA)