title: TRPV4 associates environmental temperature and sex determination in the American alligator.
abstract: Temperature-dependent sex determination (TSD), commonly found among reptiles, is a sex determination mode in which the incubation temperature during a critical temperature sensitive period (TSP) determines sexual fate of the individual rather than the individual's genotypic background. In the American alligator (Alligator mississippiensis), eggs incubated during the TSP at 33 °C (male producing temperature: MPT) yields male offspring, whereas incubation temperatures below 30 °C (female producing temperature: FPT) lead to female offspring. However, many of the details of the underlying molecular mechanism remains elusive, and the molecular link between environmental temperature and sex determination pathway is yet to be elucidated. Here we show the alligator TRPV4 ortholog (AmTRPV4) to be activated at temperatures proximate to the TSD-related temperature in alligators, and using pharmacological exposure, we show that AmTRPV4 channel activity affects gene expression patterns associated with male differentiation. This is the first experimental demonstration of a link between a well-described thermo-sensory mechanism, TRPV4 channel, and its potential role in regulation of TSD in vertebrates, shedding unique new light on the elusive TSD molecular mechanism.
authors.affiliation: Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.|Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yatsu
authors.affiliation: Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.|Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Miyagawa
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of South Carolina, and Marine Biomedicine and Environmental Science Center, Hollings Marine Laboratory, Charleston SC 29412 USA.
authors.forename: Satomi
authors.initials: S
authors.lastname: Kohno
authors.affiliation: Okazaki Institute for Integrative Bioscience, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.|Department of Physiological Sciences, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Saito
authors.affiliation: Innovative Health Applications, Kennedy Space Center, Merritt Island FL 32899 USA.
authors.forename: Russell H
authors.initials: RH
authors.lastname: Lowers
authors.affiliation: Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.|Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Ogino
authors.affiliation: Okazaki Institute for Integrative Bioscience, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Fukuta
authors.affiliation: Graduate School of Life Science and Department of Biological Sciences, Hokkaido University, Sapporo Hokkaido 062-8520 Japan.
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Katsu
authors.affiliation: Department of Veterinary Medicine, Faculty of Agriculture, Tottori University, Koyama Tottori 680-8553 Japan.
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Ohta
authors.affiliation: Okazaki Institute for Integrative Bioscience, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.|Department of Physiological Sciences, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Tominaga
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of South Carolina, and Marine Biomedicine and Environmental Science Center, Hollings Marine Laboratory, Charleston SC 29412 USA.
authors.forename: Louis J
authors.initials: LJ
authors.lastname: Guillette
authors.affiliation: Department of Basic Biology, SOKENDAI (The Graduate University for Advanced Studies), Okazaki Aichi 444-8787 Japan.|Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki Aichi 444-8787 Japan.
authors.forename: Taisen
authors.initials: T
authors.lastname: Iguchi
mesh_terms: D000489:Alligators and Crocodiles / Q000254:growth & development / Q000378:metabolism*; D000818:Animals; D001141:Aromatase / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D009095:Mullerian Ducts / Q000187:drug effects / Q000473:pathology; D009865:Oocytes / Q000378:metabolism; D018000:Receptors, Peptide / Q000235:genetics / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D019849:Sex Determination Processes / Q000187:drug effects / Q000502:physiology*; D013449:Sulfonamides / Q000494:pharmacology; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism*; D013696:Temperature; D014481:United States; D014981:Xenopus / Q000254:growth & development / Q000378:metabolism
chemical_list: C530602:N-(1-((4-(2-(((2,4-dichlorophenyl)sulfonyl)amino)-3-hydroxypropanoyl)-1-piperazinyl)carbonyl)-3-methylbutyl)-1-benzothiophene-2-carboxamide; C544743:RN 1734; D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; D055758:SOX9 Transcription Factor; D013449:Sulfonamides; D050916:TRPV Cation Channels; C084621:anti-Mullerian hormone receptor; D001141:Aromatase; D007930:Leucine; D002118:Calcium