[
International C. elegans Meeting,
2001]
A wide range of cellular processes such as muscle contraction, neurotransmitter release, secretion of hormones and cell cycling are regulated by the second messenger calcium. Intracellular calcium stores hold a key position in calcium homeostasis. These stores accumulate and release calcium in order to control cytosolic calcium levels and thus allow rapid establishment of local gradients of calcium. In addition, calcium in the lumen of the stores affects synthesis, folding, proteolytic cleavage and sorting of proteins in the endoplasmic reticulum. Removal of cytosolic calcium and filling of the stores is regulated by the activity of the Sarcoplasmic/Endoplasmic Reticulum Calcium transport ATPase (SERCA). SERCA genes are implied in a number of diseases such as Darier-White disease, Brody myopathy, cardiac hypertrophy, heart failure and type II diabetes. In higher vertebrates, three different SERCA genes exist. Protein diversity is increased by alternative splicing. The C. elegans genome contains a single SERCA gene whose transcript undergoes alternative splicing in a manner remeniscent of vertebrate SERCA2. The C. elegans and mammalian proteins show 70% identity and 80% similarity. To characterize the C. elegans SERCA gene we combined approaches in the fields of molecular biology, genetics, pharmacology and biochemistry. C. elegans SERCA cDNAs were expressed in COS cells and microsomes representing fragmented endoplasmic reticulum membranes were prepared. Phosphorylation studied showed a calcium-dependent phosphorylated intermediate, an important step in the catalytic cycle of the enzyme. Moreover, Ca 2+ -uptake experiments showed that the two isoforms have a 2-fold different affinity for Ca 2+ . Inactivation of SERCA by RNAi or gene ablation leads to embryonic or early larval lethality respectively, indicating that embryogenesis requires maternally contributed SERCA. Arrested young larvae lacking SERCA show defects in movement, pharyngeal pumping and defecation. Rescue experiments indicate an additional role for SERCA in gonadal sheath contractility. Similar defects as observed in animals lacking SERCA could be induced pharmacologically using the SERCA-specific inhibitor thapsigargin, indicating conservation of the thapsigargin-binding site. Together, these results show that SERCA is required for the function of various contractile tissues in C. elegans . This notion is supported by strong SERCA expression in all muscle types, the intestine and the gonadal myoepithelium as shown by GFP fusion constructs of both isoforms. Thus, the data presented here open a path to study SERCA function and regulation in C. elegans .
[
1987]
The concept of developmentally programmed senescence has been outlined by Leonard Hayflick (this volume), and examples from development have been used as exemplars of "developmentally programmed senescence" (Richard Russell, this volume). Unlike development, senescence has probably evolved in the absence of direct selection for increased longevity, perhaps as a direct result of the absence of such selection. (For an excellent review see Charlesworth.) A popular evolutionary model that has received experimental support suggests that senescence may result from pleiotropic effects of selection for adaptive life history characteristics. In the literature on aging, less rigorous arguements have been used to suggest that in human evolution, a delay in the age of senescence has been indirectly selected for by means of so-called longevity assurance or longevity-determinant genes. However, all explanations for the evolution of senescence are theoretical, and, with few exceptions, remain largely untested. Like Dr. Hayflick and Russell, I will assume that by developmental programming we mean genetic specification. I will use a general definition so as not to preclude examples that fail to meet one or more of the rigid criteria defined by Russell (this volume). This less rigid definition of programmed aging is necessary, because unlike development, where genetics has been successfully applied for 50 years, examples of genetic specification of senescent processes are quite few. In the literature on aging, it is still not widely accepted that mutants can alter fundamental patterns of senescent events in well-defined ways. One purpose of this presentation is to outline a few examples. In senescence, large batteries of new genes are not differentially regulated; this is quite unlike development, where many genes are differentially regulated. The molecular etiology of senescence is unknown in almost every instance and, as such, makes the study of aging a fascinating area for inquiry. If senescence is unlike development in lacking differential gene regulation, what are the approaches that are likely to yield useful results in the analysis of senescence and the aging process? The developmental genetic paradigm is a useful, indeed essential, theoretical construct for approaching the aging process in an experimental context. The lack of a suitable model organism in which classical and molecular genetics can be productively combined with other experimental techniques has impeded our understanding of senescence. Despite a general lack of evidence for genetic specification, there are instances where genetic specification is clearly evident; the analysis of mutational events that alter normal senescence in well-defined ways demonstrates this point. These instances also provide experimental models for dissecting the aging