Physiology

Larval abundance, age, growth and hatching patterns of two sympatric clingfishes, Gobiesox marmoratus and Sicyases sanguineus (Pisces, Gobiesocidae), were estimated by using otolith microstructure analysis and compared on the basis of collections performed during the austral spring in 2010 off the coast of central Chile. G. marmoratus larvae were more abundant than S. sanguineus larvae during the study period. For both species, the sagittae deposited micro-increments during embryonic development (before hatching) and a hatch mark was observable in all examined otoliths. The sagittae otoliths of G. marmoratus grew in radius, perimeter and area faster than did the otoliths of S. sanguineus. Both species showed significant (P < 0.05) differences in larval growth and lunar periodicity of the hatching events. G. marmoratus hatched at smaller sizes (2.6 mm) mainly during the first-quarter moon and the larvae grew at rates of 0.24 ± 0.01 mm day–1. S. sanguineus hatched as larger larvae ...

Patient-specific cardiovascular simulations can provide clinicians with predictive tools, fill current gaps in clinical imaging capabilities, and contribute to the fundamental understanding of disease progression. However, clinically relevant simulations must provide not only local hemodynamics, but also global physiologic response. This necessitates a dynamic coupling between the Navier–Stokes solver and reduced-order models of circulatory physiology, resulting in numerical stability and efficiency challenges. In this review, we discuss approaches to handling the coupled systems that arise from cardiovascular simulations, including recent algorithms that enable efficient large-scale simulations of the vascular system. We maintain particular focus on multiscale modeling algorithms for finite element simulations. Because these algorithms give rise to an ill-conditioned system of equations dominated by the coupled boundaries, we also discuss recent methods for solving the linear syste...

We recently reported that ANG II inhibits NBCe1 current and surface expression in Xenopus laevis oocytes (Perry C, Blaine J, Le H, and Grichtchenko II. Am J Physiol Renal Physiol 290: F417–F427, 2006). Here, we investigated mechanisms of ANG II-induced changes in NBCe1 surface expression. We showed that the PKC inhibitor GF109203X blocks and EGTA reduces surface cotransporter loss in ANG II-treated oocytes, suggesting roles for PKC and Ca2+. Using the endosomal marker FM 4-64 and enhanced green fluorescent protein (EGFP)-tagged NBCe1, we showed that ANG II stimulates endocytosis of NBCe1. To eliminate the possibility that ANG II inhibits NBCe1 recycling, we demonstrated that the recycling inhibitor monensin decreases surface expression, accumulates NBCe1-EGFP in endosomes, and inhibits NBCe1 current. Monensin and ANG II applied together produce greater inhibition of NBCe1 current than either did alone. This additive effect of monensin and ANG II suggests that ANG II stimulates inter...

We studied the extracellular [HCOabstract3−] dependence of two renal clones of the electrogenic Na/HCO3cotransporter (NBC) heterologously expressed inXenopusoocytes. We used microelectrodes to measure the change in membrane potential (ΔVm) elicited by the NBC cloned from the kidney of the salamanderAmbystoma tigrinum(akNBC) and by the NBC cloned from the kidney of rat (rkNBC). We used a two-electrode voltage clamp to measure the change in current (ΔI) elicited by rkNBC. Briefly exposing an NBC-expressing oocyte to HCOabstract3−/CO2(0.33–99 mM HCOabstract3−, pHo7.5) elicited an immediate, DIDS (4,4-diisothiocyanatostilbene-2,2-disulfonic acid)-sensitive and Na+-dependent hyperpolarization (or outward current). In ΔVmexperiments, the apparentKmfor HCOabstract3−of akNBC (10.6 mM) and rkNBC (10.8 mM) were similar. However, under voltage-clamp conditions, the apparentKmfor HCOabstract3−of rkNBC was less (6.5 mM). Because it has been reported that SOabstract3=/HSOabstract3−stimulates Na/H...

Cholinergic agonists are major stimuli for fluid secretion in parotid acinar cells. Saliva bicarbonate is essential for maintaining oral health. Electrogenic and electroneutral Na+-HCO3− cotransporters (NBCe1 and NBCn1) are abundant in parotid glands. We previously reported that angiotensin regulates NBCe1 by endocytosis in Xenopus oocytes. Here, we studied cholinergic regulation of NBCe1 and NBCn1 membrane trafficking by confocal fluorescent microscopy and surface biotinylation in parotid epithelial cells. NBCe1 and NBCn1 colocalized with E-cadherin monoclonal antibody at the basolateral membrane (BLM) in polarized ParC5 cells. Inhibition of constitutive recycling with the carboxylic ionophore monensin or the calmodulin antagonist W-13 caused NBCe1 to accumulate in early endosomes with a parallel loss from the BLM, suggesting that NBCe1 is constitutively endocytosed. Carbachol and PMA likewise caused redistribution of NBCe1 from BLM to early endosomes. The PKC inhibitor, GF-109203X...

We examined membrane trafficking of NBCe1-A and NBCe1-B variants of the electrogenic Na+-HCO3− cotransporter (NBCe1) encoded by the SLC4A4 gene, using confocal fluorescent microscopy in rat parotid acinar cells (ParC5 and ParC10). We showed that yellow fluorescent protein (YFP)-tagged NBCe1-A and green fluorescent protein (GFP)-tagged NBCe1-B are colocalized with E-cadherin in the basolateral membrane (BLM) but not with the apical membrane marker zona occludens 1 (ZO-1). We inhibited constitutive recycling with monensin and W13 and detected that NBCe1-A and NBCe1-B accumulated in vesicles marked with the early endosomal marker early endosome antigen-1 (EEA1), with a parallel loss from the BLM. We observed that NBCe1-A and NBCe1-B undergo massive carbachol (CCh)-stimulated redistribution from the BLM into early endosomes. We showed that internalization of NBCe1-A and NBCe1-B was prevented by the general PKC inhibitor GF-109203X, the PKCαβγ-specific inhibitor Gö-6976, and the PKCδ-spe...

The renal electrogenic Na+-HCO3− cotransporter (hkNBCe1) plays a major role in the bicarbonate reabsorption by the kidney. We examined how PMA- and ANG II-activated PKCs regulate hkNBCe1 expressed with or without the ANG II receptors AT1B in Xenopus laevis oocytes. We found that 10 nM PMA halved the hkNBCe1 current detected in voltage-clamped oocytes. A PKC-specific inhibitor GF-109203X, and a specific inhibitor of Ca-dependent conventional PKCαβγ, GÖ-6976, significantly reduced PMA inhibition. PMA did not alter surface expression of the cotransporters, but it significantly increased hkNBCe1-PKCαβγ membrane association. We found that at 10−6 M, ANG II halved the hkNBCe1 current detected in oocytes coexpressing cotransporters with AT1B. A PKC-specific inhibitor GF-109203X, and a PKCε translocation inhibitor εV1–2 peptide as well as BAPTA-AM (but not GÖ-6976), significantly reduced ANG II inhibition. At 10−6 M, ANG II significantly decreased surface expression of the cotransporters an...

Due to the limited availability of suitable positron emission tomography (PET) tracers, the majority of myocardial perfusion imaging (MPI) scans is performed using SPECT rather than PET. The aim of this study is to design and synthesize carbon-11-labeled ammonium salt derivatives and explore their structure–activity relationship (SAR) and their potential as PET–MPI agents. Three carbon-11-labeled ammonium salts were developed. SAR of the labeled compounds were explored vis-à-vis the effects of charge density and lipophilicity on the distribution kinetics in mice. These studies pointed at [11C]4 as the lead compound. Comparative microPET/CT scans in healthy rats, using both [11C]4 and [13 N]–NH3, substantiated the potential of [11C]4 ([11C]-DMDPA). A proof of concept for the potential of radiolabeled ammonium salts as MPI agents has been demonstrated in a newly developed swine model of permanent partial coronary artery occlusion. SAR studies of 11C-labeled ammonium salts suggest that both lipophilicity and charge density affect the performance of these compounds as MPI probes. In a swine model, the labeled lead successfully visualized the defect regions in the myocardium. The data presented call for the development of fluorine-18 analogues, to increase clinical impact.