TRP Channels

Chapter 12Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 16:56:48

TRPML2 and the Evolution of Mucolipins

Emma N. Flores and Jaime García-Añoveros

Abstract TRPML2, the polypeptide product of the gene Trpml2 (aka Mcoln2), is a member of the TRPML or mucolipin branch of the TRP super family of ion channels. Although no known agonists have been discovered, the wild type channel gives basal currents when heterologously expressed in Drosophila (S2) cells and is constitutively active inmammalian cells when bearing a cell degeneration-causing, proline to alanine substitution in the fifth trans-membrane domain. TRPML2 forms channels that are inwardly rectifying and permeable to Ca+2, Na+, and Fe+2. Localization studies indicate TRPML2 is present in lysosomes, late endosomes, recycling endosomes and, at a lower level, the plasma membrane. Tissue and organ distribution of TRPML2 is solely reported through RT-PCR and it is uncertain which cell types express this channel. However, various studies suggest that lymphoid cells express TRPML2. Although the function of TRPML2 is not known, distribution and channel properties suggest it could play roles in calcium release from endolysosomes, perhaps to mediate calcium-dependent events such as vesicle fusion, or to release calcium from intracellular acidic stores. However, TRPML2 may also function in the plasma membrane and its abundance in vesicles of the endocytic pathaway might occur because its presence in the cell surface is regulated by endocytosis and exocytosis. An evolutionary analysis of Trpml2 and its relatives reveals that vertebrate and invertebrate chordates have only one Trpml gene, that Trpml1 and Trpml2 are common to vertebrates, and that Trpml3 is only found in tetrapods. Ray-finned fishes contain another isoform, which we term Trpml4 or Mcoln4 (and its product TRPML4). Trpml2 is next to Trpml3 in all tetrapod genomes except that of the frog Xenopus tropicalis and of the domesticated pig, which seems to lack most of the Trpml3 gene. This close linkage across species implies that it is maintained by selective pressure and suggests that the regulation of both genes is interdependent.

Chapter 13Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 16:53:25

The TRPML3 Channel: From Gene to Function

Konrad Noben-Trauth

Abstract TRPML3 is a transient receptor potential (TRP) channel that is encoded by the mucolipin 3 gene (MCOLN3), a member of the small mucolipin gene family. Mcoln3 shows a broad expression pattern in embryonic and adult tissues that includes differentiated cells of skin and inner ear. Dominant mutant alleles of murine Mcoln3 cause embryonic lethality, pigmentation defects and deafness. The TRPML3 protein features a six-transmembrane topology and functions as a Ca2+ permeable inward rectifying cation channel that is open at sub-physiological pH and closes as the extracytosolic pH becomes more acidic. TRPML3 localizes to the plasmamembrane and to early- and late-endosomes as well as lysosomes. Recent advances suggest that TRPML3 may regulate the acidification of early endosomes, hence playing a critical role in the endocytic pathway.

Chapter 14Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 15:14:52

TRPV5 and TRPV6 in Transcellular Ca2+ Transport: Regulation, Gene Duplication, and Polymorphisms in African Populations

Ji-Bin Peng

TRPV5 and TRPV6 are unique members of the TRP super family. They are highly selective for Ca2+ ions with multiple layers of Ca2+-dependent inactivation mechanisms, expressed at the apical membrane of Ca2+ transporting epithelia, and robustly responsive to 1,25-dihydroxivitamin D3. These features are well suited for their roles as Ca2+ entry channels in the first step of transcellular Ca2+ transport pathways, which are involved in intestinal absorption, renal reabsorption of Ca2+, placental transfer of Ca2+ to fetus, and many other processes. While TRPV6 is more broadly expressed in a variety of tissues such as esophagus, stomach, small intestine, colon, kidney, placenta, pancreas, prostate, uterus, salivary gland, and sweat gland, TRPV5 expression is relatively restricted to the distal convoluted tubule and connecting tubule of the kidney. There is only one TRPV6-like gene in fish and birds in comparison to both TRPV5 and TRPV6 genes in mammals, indicating TRPV5 gene was likely generated from duplication of TRPV6 gene during the evolution of mammals to meet the needs of complex renal function. TRPV5 and TRPV6 are subjected to vigorous regulations under physiological, pathological, and therapeutic conditions. The elevated TRPV6 level in malignant tumors such as prostate and breast cancers makes it a potential therapeutic target. TRPV6, and to a lesser extent TRPV5, exhibit unusually high levels of single nucleotide polymorphisms (SNPs) in African populations as compared to other populations, indicating TRPV6 gene was under selective pressure during or after humans migrated out of Africa. The SNPs of TRPV6 and TRPV5 likely contribute to the Ca2+ conservation mechanisms in African populations.

Chapter 15Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 12:36:40

The TRPV5 Promoter as a Tool for Generation of Transgenic Mouse Models

Marlene Vind Hofmeister, Ernst-Martin Füchtbauer, Robert Andrew Fenton, and Jeppe Praetorius

The transient receptor potential vanilloid 5 (TRPV5) is a Ca2+ channel, which is expressed in renal late distal convoluted tubules (DCT2s) and connecting tubules (CNTs). These tubules play a major role in hormone controlled renal Ca2+ reabsorption, and thereby in body Ca2+ homeostasis, as well as urinary excretion of other electrolytes, including Na+ and K+. DCT2 and CNT are difficult to distinguish from the surrounding structures and thereby to study by direct functional methods. We developed a transgenic mouse model expressing enhanced green fluorescent protein (EGFP) driven by the TRPV5 promoter to identify these specific tubules. Expression of EGFP in the DCT2 and CNT allows the isolation of pure DCT2 and CNT populations for proteomic and physiological analyses. The TRPV5 promoter is also useful for generating conditional knockout mouse models in a cellspecific manner. TRPV5 promoter driven Cre recombinase expression will be useful for inducing DCT2 and CNT specific gene silencing of various channels, pumps, carriers, and receptors. In this chapter, we describe the strategy for developing transgenic mouse lines involving the TRPV5 promoter, provide a description of extensive validation of these mouse lines, and discuss possible uses and limitations.

Chapter 16Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 12:33:16

TRPP Channels and Polycystins

Alexis Hofherr and Michael Köttgen

The founding member of the TRPP family, TRPP2, was identified as one of the disease genes causing autosomal dominant polycystic kidney disease (ADPKD). ADPKD is the most prevalent, potentially lethal, monogenic disorder in humans, with an average incidence of one in 400 to one in 1,000 individuals worldwide. Here we give an overview of TRPP ion channels and Polycystin-1 receptor proteins focusing on more recent studies. We include the Polycystin-1 family since these proteins are functionally linked to TRPP channels

Chapter 17Abstracts

Posted by Md. Shahidul Islam Wed, February 16, 2011 12:25:40

TRP Channels in Yeast

Marta Kaleta and Christopher Palmer

Microbes have made numerous contributions to the study of biology and medicine. Those contributions also include many original discovery’s in the study of ion channels often thought as the province of neuroscientists or cardiophysiologists. Yeast have long been used as a model organism and TRP channel genes and their transmembrane products touted as the "vanguards of the sensory system" can be identified in the genomes of many yeasts. This article aims to review the study of these TRP channels in yeast their discovery, electrophysiological properties and physiological function.

Chapter 18Abstracts

Posted by Md. Shahidul Islam Mon, February 07, 2011 19:45:23

C. elegans TRP Channels

Rui Xiao and X.Z. Shawn Xu

Transient receptor potential (TRP) channels represent a superfamily of cation channels found in all eukaryotes. The C. elegans genome encodes seventeen TRP channels covering all of the seven TRP subfamilies. Genetic analyses in C. elegans have implicated TRP channels in a wide spectrum of behavioral and physiological processes, ranging from sensory transduction (e.g. chemosensation, touch sensation, proprioception and osmosensation) to fertilization, drug dependence, organelle biogenesis, apoptosis, gene expression, and neurotransmitter/hormone release. Many C. elegans TRP channels share similar activation and regulatory mechanisms with their vertebrate counterparts. Studies in C. elegans have also revealed some previously unrecognized functions and regulatory mechanisms of TRP channels. C. elegans represents an excellent genetic model organism for the study of function and regulation of TRP channels in vivo.

Chapter 19Abstracts

Posted by Md. Shahidul Islam Mon, February 07, 2011 19:31:14

Investigations of the In Vivo Requirements of Transient Receptor Potential Ion Channels Using Frog and Zebrafish Model Systems

Robert A. Cornell

Transient Receptor Potential (TRP) channels are cation channels that serve as cellular sensors on the plasma membrane, and have other less-well defined roles in intracellular compartments. The first TRP channel was identified upon molecular characterization of a fly mutant with abnormal photoreceptor function. More than 20 TRP channels have since been identified in vertebrates and invertebrate model systems, and these are divided into subfamilies based on structural similarities. The biophysical properties of TRP channels have primarily been explored in tissue culture models. The in vivo requirements for TRPs have been studied in invertebrate models like worm and flies, and also in vertebrate models, primarily mice and rats. Frog and zebrafish model systems offer certain experimental advantages relative to mammalian systems, and here a selection of papers which capitalize on these advantages to explore vertebrate TRP channel biology are reviewed. For instance, frog oocytes are useful for biochemistry and for electrophysiology, and these features were exploited in the identifcation TRPC1 as a candidate vertebrate mechanoreceptor. Also, the spinal neurons from frog embryos can be readily grown in culture. This feature was used to establish a role for TRPC1 in axon pathfinding in these neurons, and to explore how TRPC1 activity is regulated in this context. Zebrafish embryos are transparent making them well suited for in vivo imaging studies. This quality was exploited in a study in which the trpc2 gene promoter was used to label and trace the axon pathway of a subset of olfactory sensory neurons. Another experimental advantage of zebrafish is the speed and low cost of manipulating gene expression in embryos. Using these methods, it has been shown that TRPN1 is necessary for mechanosensation in zebrafish hair cells. Frogs and fish genomes have been mined to make inferences regarding evolutionary diversification of the thermosensitive TRP channels. Finally, TRPM7 is required for early morphogenesis in mice but not in fish; the reason for this difference is unclear, but it has caused zebrafish to be favored for exploration of TRPM7’s role in later events in embryogenesis. The special experimental attributes of frogs and zebrafish suggest that these animals will continue to play an important role as models in future explorations of TRP channel biology.