Annual Review of Physiology - Volume 70, 2008

This article mainly presents, in sequential panels of time, an overview of my professional involvements and laboratory experiences. I became smitten with red blood cells early on, and this passion remains with me to this day. I highlight certain studies, together with those who performed the work, recognizing that it was necessary to limit the details and the topics chosen for discussion. I am uncertain of the interest a personal account has for others, but at least it's here for the record.

Calcium (Ca) is a universal intracellular second messenger. In muscle, Ca is best known for its role in contractile activation. However, in recent years the critical role of Ca in other myocyte processes has become increasingly clear. This review focuses on Ca signaling in cardiac myocytes as pertaining to electrophysiology (including action potentials and arrhythmias), excitation-contraction coupling, modulation of contractile function, energy supply-demand balance (including mitochondrial function), cell death, and transcription regulation. Importantly, although such diverse Ca-dependent regulations occur simultaneously in a cell, the cell can distinguish distinct signals by local Ca or protein complexes and differential Ca signal integration.

Low oxygen (O2) levels are a naturally occurring feature of embryonic development, adult physiology, and diseases such as those of the cardiovascular system. Although many responses to O2 deprivation are mediated by hypoxia-inducible factors (HIFs), researchers are finding a growing number of HIF-independent pathways that promote O2 conformance and hypoxia tolerance. Here, we describe HIF-independent responses and how they impact cardiovascular tissue homeostasis.

Bcl-2 family members are important regulators of cell survival and cell death. Researchers have focused mainly on mitochondria, where both proapoptotic and antiapoptotic family members function to regulate the release of cytochrome c and other mediators of apoptosis. However, as reviewed here, Bcl-2 family members also operate on another front, the endoplasmic reticulum (ER), to both positively and negatively regulate the release of Ca²⁺. There is abundant evidence that Ca²⁺ signals trigger apoptosis in response to a wide variety of agents and conditions. Conversely, Ca²⁺ signals can also mediate cell survival. Recent findings indicate that Bcl-2 interacts with inositol 1,4,5-trisphosphate (IP3) receptor Ca²⁺ channels on the ER, regulating their opening in response to IP3 and thus inhibiting IP3-mediated Ca²⁺ signals that induce apoptosis while enhancing Ca²⁺ signals that support cell survival.

Sperm are attracted by chemical factors that are released by the egg—a process called chemotaxis. Most of our knowledge on sperm chemotaxis originates from the study of marine invertebrates. In recent years, the main features of the chemotactic signaling pathway and the swimming behavior evoked by chemoattractants have been elucidated in sea urchins. In contrast, our understanding of mammalian sperm chemotaxis is still rudimentary and subject to an ongoing debate. In this review, we raise new questions and discuss current concepts of sperm chemotaxis. Finally, we highlight commonalities and differences of sensory signaling in sperm, photoreceptors, and olfactory neurons.

Studies of the physiology and biomechanics of small (∼1 cm) organisms are often limited by the inability to see inside the animal during a behavior or process of interest and by a lack of three-dimensional morphology at the submillimeter scale. These constraints can be overcome by an imaging probe that has sensitivity to soft tissue, the ability to penetrate opaque surfaces, and high spatial and temporal resolution. Synchrotron X-ray imaging has been successfully used to visualize millimeter-centimeter-sized organisms with micrometer-range spatial resolutions in fixed and living specimens. Synchrotron imaging of small organisms has been the key to recent novel insights into structure and function, particularly in the area of respiratory physiology and function of insects. X-ray imaging has been effectively used to examine the morphology of tracheal systems, the mechanisms of tracheal and air sac compression in insects, and the function of both chewing and sucking mouthparts in insects. Synchrotron X-ray imaging provides an exciting new window into the internal workings of small animals, with future promise to contribute to a range of physiological and biomechanical questions in comparative biology.

Since the time of Muybridge and Marey in the last half of the nineteenth century, studies of animal movement have relied on some form of high-speed or stop-action imaging to permit analysis of appendage and body motion. In the past ten years, the advent of megapixel-resolution high-speed digital imaging with maximal framing rates of 250 to 100,000 images per second has allowed new views of musculoskeletal function in comparative physiology that now extend to imaging flow around moving animals and the calculation of fluid forces produced by animals moving in fluids. In particular, the technique of digital particle image velocimetry (DPIV) has revolutionized our ability to understand how moving animals generate fluid forces and propel themselves through air and water. DPIV algorithms generate a matrix of velocity vectors through the use of image cross-correlation, which can then be used to calculate the force exerted on the fluid as well as locomotor work and power. DPIV algorithms can also be applied to images of moving animals to calculate the velocity of different regions of the moving animal, providing a much more detailed picture of animal motion than can traditional digitizing methods. Although three-dimensional measurement of animal motion is now routine, in the near future model-based kinematic reconstructions and volumetric analyses of animal-generated fluid flow patterns will provide the next step in imaging animal biomechanics and physiology.

Steroids play an important role in the regulation of normal physiology and the treatment of disease. Steroid receptors have classically been described as ligand-activated transcription factors mediating long-term genomic effects in hormonally regulated tissues. It is now clear that steroids also mediate rapid signaling events traditionally associated with growth factor receptors and G protein–coupled receptors. Although evidence suggests that the classical steroid receptors are capable of mediating many of these events, more recent discoveries reveal the existence of transmembrane receptors capable of responding to steroids with cellular activation. One such receptor, GPR30, is a member of the G protein–coupled receptor superfamily and mediates estrogen-dependent kinase activation as well as transcriptional responses. In this review, we provide an overview of the evidence for the cellular and physiological actions of GPR30 in estrogen-dependent processes and discuss the relationship of GPR30 with classical estrogen receptors.

Insulin-like signaling is critical for nutrient homeostasis, growth and survival. However, work with lower metazoans—Caenorhabditis elegans and Drosophila—shows that reduced insulin-like signaling extends life span. In addition, reduced insulin signaling in higher animals—rodents and humans—causes glucose intolerance and hyperinsulinemia that progresses to diabetes and shortens the life span of affected individuals. Hyperinsulinemia usually develops to maintain glucose homeostasis and prevent the progression toward life-threatening type 2 diabetes; however, increased circulating insulin may have negative effects on the brain that promote age-related disease. We discuss the possibility that the brain is the site where reduced insulin-like signaling can consistently extend mammalian life span—just as reduced insulin-like signaling extends the life span of lower metazoans.

Neurons that produce gonadotropin-releasing hormone (GnRH) reside in the basal forebrain and drive reproductive function in mammals. Understanding the circuitry that regulates GnRH neurons is fundamental to comprehending the neuroendocrine control of puberty and reproduction in the adult. This review focuses on a family of neuropeptides encoded by the Kiss1 gene, the kisspeptins, and their cognate receptor, GPR54, which have been implicated in the regulation of GnRH secretion. Kisspeptins are potent secretagogues for GnRH, and the Kiss1 gene is a target for regulation by gonadal steroids (e.g., estradiol and testosterone), metabolic factors (e.g., leptin), photoperiod, and season. Kiss1 neurons in the arcuate nucleus may regulate the negative feedback effect of gonadal steroids on GnRH and gonadotropin secretion in both sexes. The expression of Kiss1 in the anteroventral periventricular nucleus (AVPV) is sexually dimorphic, and Kiss1 neurons in the AVPV may participate in the generation of the preovulatory GnRH/luteinizing hormone (LH) surge in the female rodent. Kiss1 neurons have emerged as primary transducers of internal and environmental cues to regulate the neuroendocrine reproductive axis.

The increasing prevalence of obesity worldwide has imparted renewed impetus to the study of the mechanisms of appetite regulation. Digestion and nutrient absorption take place in the gastrointestinal (GI) tract, whereas food intake is controlled by neuronal circuits in the central nervous system. The need for gut-brain cross talk is therefore clear. It is now recognized that hormones released into the circulation from the GI tract in response to nutritional stimuli form a key component of this gut-brain axis. Peptides such as glucagon-like peptide-1, oxyntomodulin, pancreatic polypeptide, and peptide YY3–36 reduce food intake in both animal models and in humans. Physiologically, such peptides are thought to act as satiety signals and meal terminators. Here, we review the current state of the field of the effects of gut hormone action on appetite control.

Ca²⁺ is essential for numerous physiological functions in our bodies. Therefore, its homeostasis is finely maintained through the coordination of intestinal absorption, renal reabsorption, and bone resorption. The Ca²⁺-selective epithelial channels TRPV5 and TRPV6 have been identified, and their physiological roles have been revealed: TRPV5 is important in final renal Ca²⁺ reabsorption, and TRPV6 has a key role in intestinal Ca²⁺ absorption. The TRPV5 knockout mice exhibit renal leak hypercalciuria and accordingly upregulate their intestinal TRPV6 expression to compensate for their negative Ca²⁺ balance. In contrast, despite their severe negative Ca²⁺ balance, TRPV6-null mice do not display any compensatory mechanism, thus resulting in secondary hyperparathyroidism. These results indicate that the genes for TRPV5 and TRPV6 are differentially regulated in human diseases associated with disturbed Ca²⁺ balance such as hypercalciuria, osteoporosis, and vitamin D–resistant rickets.

Cytosolic Ca²⁺ signals are crucial for the control of fluid and enzyme secretion from exocrine glands. The highly polarized exocrine acinar cells have evolved sophisticated and complex Ca²⁺ signaling mechanisms that exercise precise control of the secretory events occurring across the apical plasma membrane bordering the gland lumen. Ca²⁺ stores in the endoplasmic reticulum, the secretory granules, the lysosomes, and the endosomes all play important roles in the generation of the local apical Ca²⁺ spikes that switch on Cl⁻ channels in the apical plasma membrane as well as exocytotic export of enzymes. The mitochondria are crucial not only for ATP generation but also for the physiologically important subcellular compartmentalization of the cytosolic Ca²⁺ signals.

The discovery of aquaporin water channels by Agre and coworkers answered a long-standing biophysical question of how the majority of water crosses biological membranes. The identification and study of aquaporins have provided insight, at the molecular level, into the fundamental physiology of water balance regulation and the pathophysiology of water balance disorders. In addition to the originally identified classical aquaporins, a second class of aquaporins has been identified. Aquaporins in this latter class, the so-called aquaglyceroporins, transport small uncharged molecules such as glycerol and urea as well as water. Aquaglyceroporins have a wide tissue distribution, and emerging data suggest that several of them may play previously unappreciated physiological or pathophysiological roles. Analyses of transgenic mice have revealed potential roles of aquaglyceroporins in skin elasticity, gastrointestinal function and metabolism, and metabolic diseases such as diabetes mellitus. This review comprehensively discusses the recent discoveries in the field of aquaglyceroporins, alongside a brief overview of the so-called unorthodox aquaporins.

Mutations in the serine-threonine kinases WNK1 and WNK4 cause a Mendelian disease featuring hypertension and hyperkalemia. In vitro and in vivo studies have revealed that these proteins are molecular switches that have discrete functional states that impart different effects on downstream ion channels, transporters, and the paracellular pathway. These effects enable the distal nephron to allow either maximal NaCl reabsorption or maximal K⁺ secretion in response to hypovolemia or hyperkalemia, respectively. The related kinase WNK3 has reciprocal actions on the primary mediators of cellular Cl⁻ influx and efflux, effects that can serve to regulate cell volume during growth and in response to osmotic stress as well as to modulate neuronal responses to GABA. These findings define a versatile new family of kinases that coordinate the activities of diverse ion transport pathways to achieve and maintain fluid and electrolyte homeostasis.

Cyclooxygenase-derived prostanoids exert complex and diverse functions within the kidney. The biological effect of each prostanoid is controlled at multiple levels, including (a) enzymatic reactions catalyzed sequentially by cyclooxygenase and prostanoid synthase for the synthesis of bioactive prostanoid and (b) the interaction with its receptors that mediate its functions. Cyclooxygenase-derived prostanoids act in an autocrine or a paracrine fashion and can serve as physiological buffers, protecting the kidney from excessive functional changes during physiological stress. Through these actions, prostanoids play important roles in maintaining renal function, body fluid homeostasis, and blood pressure. Renal cortical COX2-derived prostanoids, particularly PGI2 and PGE2, play critical roles in maintaining blood pressure and renal function in volume-contracted states. Renal medullary COX2-derived prostanoids appear to have an antihypertensive effect in individuals challenged with a high-salt diet. Loss of EP2 or IP receptor is associated with salt-sensitive hypertension. COX2 also plays a role in maintaining renal medullary interstitial cell viability in the hypertonic environment of the medulla. Cyclooxygenase-derived prostanoids also are involved in certain pathological processes. The cortical COX2-derived PGI2 participates in the pathogenesis of renal vascular hypertension through stimulating renal renin synthesis and release. COX-derived prostanoids also appear to be involved in the pathogenesis of diabetic nephropathy. COXs, prostanoid synthases, and prostanoid receptors should provide fruitful targets for intervention in the pharmacological treatment of renal disease.

The concept of a regulatory link between the gastrointestinal tract and kidneys is not new. The idea that dietary intake and composition can affect renal function is perhaps self-evident, but defining this relationship, especially in terms of sensors and effectors, is proving more difficult. That the gastrointestinal tract can exert some control over renal function was strengthened by the early observation that oral ingestion of a sodium chloride load has a greater natriuretic effect than when the same amount is given intravenously. This effect was subsequently shown to be independent of changes in aldosterone and atrial natriuretic peptide, although not necessarily angiotensin-II. However, the nature of any intestinal natriuretic peptide remains uncertain, despite suggestions that various gut-derived hormones, including guanylin and uroguanylin, may be involved. There is also an emerging concept of gastrointestinal taste and taste-like receptor mechanisms present throughout the gastrointestinal tract, which may govern the excretion of other key electrolytes, including potassium and phosphate. The evidence for gut sensors of nutrients such as proteins, amino acids, glucose, and acid is now becoming more established. Thus, we can anticipate the existence and eventual characterization of several gut ion sensors.

Mucins are important components that exert a variety of functions in cell-cell interaction, epidermal growth factor receptor signaling, and airways protection. In the conducting airways of the lungs, mucins are the major contributor to the viscoelastic property of mucous secretion, which is the major barrier to trapping inhaled microbial organism, particulates, and oxidative pollutants. The homeostasis of mucin production is an important feature in conducting airways for the maintenance of mucociliary function. Aberrant mucin secretion and accumulation in airway lumen are clinical hallmarks associated with various lung diseases, such as asthma, chronic obstructive pulmonary disease, cystic fibrosis, emphysema, and lung cancer. Among 20 known mucin genes identified, 11 of them have been verified at either the mRNA and/or protein level in airways. The regulation of mucin genes is complicated, as are the mediators and signaling pathways. This review summarizes the current view on the mediators, the signaling pathways, and the transcriptional units that are involved in the regulation of airway mucin gene expression. In addition, we also point out essential features of epigenetic mechanisms for the regulation of these genes.

Cell surface mucins are large transmembrane glycoproteins involved in diverse functions ranging from shielding the airway epithelium against pathogenic infection to regulating cellular signaling and transcription. Although hampered by the relatively recent characterization of cell surface mucins and the difficulties inherent in working with molecules of their size, numerous studies have placed the tethered mucins in the thick of normal and diseased lung physiology. This review focuses on the three best-characterized cell surface mucins expressed in the respiratory tract: MUC1, MUC4, and MUC16.