A Soluble Platelet-Derived Growth Factor Receptor-ß Originates via Pre-mRNA Splicing in the Healthy Brain and Is Upregulated during Hypoxia and Aging.

The platelet-derived growth factor-BB (PDGF-BB) pathway provides critical regulation of cerebrovascular pericytes, orchestrating their investment and retention within the brain microcirculation. Dysregulated PDGF Receptor-beta (PDGFRß) signaling can lead to pericyte defects that compromise blood-brain barrier (BBB) integrity and cerebral perfusion, impairing neuronal activity and viability, which fuels cognitive and memory deficits. Receptor tyrosine kinases such as PDGF-BB and vascular endothelial growth factor-A (VEGF-A) are often modulated by soluble isoforms of cognate receptors that establish signaling activity within a physiological range. Soluble PDGFRß (sPDGFRß) isoforms have been reported to form by enzymatic cleavage from cerebrovascular mural cells, and pericytes in particular, largely under pathological conditions. However, pre-mRNA alternative splicing has not been widely explored as a possible mechanism for generating sPDGFRß variants, and specifically during tissue homeostasis. Here, we found sPDGFRß protein in the murine brain and other tissues under normal, physiological conditions. Utilizing brain samples for follow-on analysis, we identified mRNA sequences corresponding to sPDGFRß isoforms, which facilitated construction of predicted protein structures and related amino acid sequences. Human cell lines yielded comparable sequences and protein model predictions. Retention of ligand binding capacity was confirmed for sPDGFRß by co-immunoprecipitation. Visualizing fluorescently labeled sPDGFRß transcripts revealed a spatial distribution corresponding to murine brain pericytes alongside cerebrovascular endothelium. Soluble PDGFRß protein was detected throughout the brain parenchyma in distinct regions, such as along the lateral ventricles, with signals also found more broadly adjacent to cerebral microvessels consistent with pericyte labeling. To better understand how sPDGFRß variants might be regulated, we found elevated transcript and protein levels in the murine brain with age, and acute hypoxia increased sPDGFRß variant transcripts in a cell-based model of intact vessels. Our findings indicate that soluble isoforms of PDGFRß likely arise from pre-mRNA alternative splicing, in addition to enzymatic cleavage mechanisms, and these variants exist under normal physiological conditions. Follow-on studies will be needed to establish potential roles for sPDGFRß in regulating PDGF-BB signaling to maintain pericyte quiescence, BBB integrity, and cerebral perfusion-critical processes underlying neuronal health and function, and in turn, memory and cognition.

A Soluble Platelet-Derived Growth Factor Receptor-ß Originates via Pre-mRNA Splicing in the Healthy Brain and is Differentially Regulated during Hypoxia and Aging.

The platelet-derived growth factor-BB (PDGF-BB) pathway provides critical regulation of cerebrovascular pericytes, orchestrating their investment and retention within the brain microcirculation. Dysregulated PDGF Receptor-beta (PDGFRß) signaling can lead to pericyte defects that compromise blood-brain barrier (BBB) integrity and cerebral perfusion, impairing neuronal activity and viability, which fuels cognitive and memory deficits. Receptor tyrosine kinases (RTKs) like PDGF-BB and vascular endothelial growth factor-A (VEGF-A) are often modulated by soluble isoforms of cognate receptors that establish signaling activity within a physiological range. Soluble PDGFRß (sPDGFRß) isoforms have been reported to form by enzymatic cleavage from cerebrovascular mural cells, and pericytes in particular, largely under pathological conditions. However, pre-mRNA alternative splicing has not been widely explored as a possible mechanism for generating sPDGFRß variants, and specifically during tissue homeostasis. Here, we found sPDGFRß protein in the murine brain and other tissues under normal, physiological conditions. Utilizing brain samples for follow-on analysis, we identified mRNA sequences corresponding to sPDGFRß isoforms, which facilitated construction of predicted protein structures and related amino acid sequences. Human cell lines yielded comparable sequences and protein model predictions. Retention of ligand binding capacity was confirmed for sPDGFRß by co-immunoprecipitation. Visualizing fluorescently labeled sPDGFRß transcripts revealed a spatial distribution corresponding to murine brain pericytes alongside cerebrovascular endothelium. Soluble PDGFRß protein was detected throughout the brain parenchyma in distinct regions such as along the lateral ventricles, with signals also found more broadly adjacent to cerebral microvessels consistent with pericyte labeling. To better understand how sPDGFRß variants might be regulated, we found elevated transcript and protein levels in the murine brain with age, and acute hypoxia increased sPDGFRß variant transcripts in a cell-based model of intact vessels. Our findings indicate that soluble isoforms of PDGFRß likely arise from pre-mRNA alternative splicing, in addition to enzymatic cleavage mechanisms, and these variants exist under normal physiological conditions. Follow-on studies will be needed to establish potential roles for sPDGFRß in regulating PDGF-BB signaling to maintain pericyte quiescence, BBB integrity, and cerebral perfusion - critical processes underlying neuronal health and function, and in turn memory and cognition.

RZ Interval as an Impedance Cardiography Indicator of Effort-Related Cardiac Sympathetic Activity.

Research on effort and motivation commonly assesses how the sympathetic branch of the autonomic nervous system affects the cardiovascular system. The cardiac pre-ejection period (PEP), assessed via impedance cardiography, is a common outcome, but assessing PEP requires identifying subtle points on cardiac waveforms. The present research examined the psychometric value of the RZ interval (RZ), which has recently been proposed as an indicator of sympathetic activity, for effort research. Also known as the initial systolic time interval (ISTI), RZ is the time (in ms) between the ECG R peak and the dZ/dt Z peak. Unlike PEP, RZ involves salient waveform points that are easily and reliably identified. Data from two experiments evaluated the suitability of RZ for effort paradigms and compared it to a popular automated PEP method. In Studies 1 (n = 89) and 2 (n = 71), participants completed a standard appetitive task in which each correct response earned a small amount of cash. As expected, incentives significantly affected PEP and RZ in both experiments. PEP and RZ were highly correlated (all rs ≥ 0.89), and RZ consistently yielded a larger effect size than PEP. In Study 3, a quantitative synthesis of the experiments indicated that the effect size of RZ's response to incentives (Hedges's g = 0.432 [0.310, 0.554]) was roughly 15% larger than PEP's effect size (g = 0.376 [0.256, 0.496]). RZ thus appears promising for future research on sympathetic aspects of effort-related cardiac activity.

Do depressive symptoms "blunt" effort? An analysis of cardiac engagement and withdrawal for an increasingly difficult task.

Research on depression and effort has suggested "depressive blunting"-lower cardiovascular reactivity in response to challenges and stressors. Many studies, however, find null effects or higher reactivity. The present research draws upon motivational intensity theory, a broad model of effort that predicts cases in which depressive symptoms should increase or decrease effort. Because depressive symptoms can influence task-difficulty appraisals-people see tasks as subjectively harder-people high in depressive symptoms should engage higher effort at objectively easier levels of difficulty but also quit sooner. A sample of adults completed a mental effort challenge with four levels of difficulty, from very easy to difficult-but-feasible. Depressive symptoms were assessed with the CESD and DASS; effort-related cardiac activity was assessed via markers of contractility (e.g., the cardiac pre-ejection period [PEP]) obtained with impedance cardiography. The findings supported the theory's predictions. When the task was relatively easier, people high in depressive symptoms showed higher contractility (shorter PEP), consistent with greater effort. When the task was relatively harder, people high in depressive symptoms showed diminished contractility, consistent with quitting. The results suggest that past research has been observing a small part of a larger trajectory of trying and quitting, and they illustrate the value of a theoretically grounded analysis of depressive symptoms and effort-related cardiac activity.