Project Grant 2232891

ANT LIA: THE ROLE OF SEX DETERMINATION IN THE RADIATION OF ANTARCTIC NOTOTHENIOID FISH -ANTARCTIC ANIMALS FACE TREMENDOUS THREATS AS ANTARCTIC ICE SHEETS MELT AND TEMPERATURES RISE. ABOUT 34 MILLION YEARS AGO, WHEN ANTARCTICA BEGAN TO COOL, MOST SPECIES OF FISH BECAME LOCALLY EXTINCT. A GROUP CALLED THE NOTOTHENIOIDS, HOWEVER, SURVIVED DUE TO THE EVOLUTION OF ANTIFREEZE. THE GROUP EVENTUALLY SPLIT INTO OVER 120 SPECIES. WHY DID THIS GROUP OF ANTARCTIC FISHES EVOLVE INTO SO MANY SPECIES? ONE POSSIBLE REASON WHY A SINGLE POPULATION SPLITS INTO TWO SPECIES RELATES TO SEX GENES AND SEX CHROMOSOMES. DIVERGING SPECIES OFTEN HAVE EITHER DIFFERENT SEX DETERMINING GENES (GENES THAT SPECIFY WHETHER AN INDIVIDUAL?S GONADS BECOME OVARIES OR TESTES) OR HAVE DIFFERENT SEX CHROMOSOMES (CHROMOSOMES THAT DIFFER BETWEEN MALES AND FEMALES WITHIN A SPECIES, LIKE THE HUMAN X AND Y CHROMOSOMES). WE KNOW THE SEX CHROMOSOMES OF ONLY A FEW NOTOTHENIOID SPECIES AND KNOW THE GENETIC BASIS FOR SEX DETERMINATION IN NONE OF THEM. THE AIMS OF THIS RESEARCH ARE TO: 1) IDENTIFY SEX CHROMOSOMES IN SPECIES REPRESENTING EVERY MAJOR GROUP OF ANTARCTIC NOTOTHENIOID FISH; 2) DISCOVER POSSIBLE SEX DETERMINING GENES IN EVERY MAJOR GROUP OF ANTARCTIC NOTOTHENIOID FISH; AND 3) FIND SEX CHROMOSOMES AND POSSIBLE SEX DETERMINING GENES IN TWO GROUPS OF TEMPERATE, WARMER WATER, NOTOTHENIOID FISH. THESE WARMER WATER FISH INCLUDE GROUPS THAT NEVER EXPERIENCED THE FRIGID SOUTHERN OCEAN AND GROUPS THAT HAD ANCESTORS INHABITING ANTARCTIC OCEANS THAT LATER ADJUSTED TO WARMER WATERS. THIS PROJECT WILL HELP EXPLAIN THE MECHANISMS THAT LED TO THE DIVISION OF A GROUP OF SPECIES THREATENED BY CLIMATE CHANGE. THIS INFORMATION IS CRITICAL TO CONSERVE DECLINING POPULATIONS OF ANTARCTIC NOTOTHENIOIDS, WHICH ARE MAJOR FOOD SOURCES FOR OTHER ANTARCTIC SPECIES SUCH AS BIRD AND SEALS. THE PROJECT WILL OFFER A DIVERSE GROUP OF UNDERGRADUATES THE OPPORTUNITY TO DEVELOP A PERMANENT EXHIBIT AT THE EUGENE SCIENCE CENTER MUSEUM. THE EXHIBIT WILL DESCRIBE THE ANTARCTIC ENVIRONMENT AND EXPLAIN ITS RAPID CLIMATE CHANGE. IT WILL ALSO INTRODUCE THE CONTINENT?S BIZARRE FISHES THAT LIVE BELOW THE FREEZING POINT OF WATER. THE PROJECT WILL COLLABORATE WITH THE UNIVERSITY?S SCIENCE AND COMICS INITIATIVE AND STUDENTS IN THE ENGLISH DEPARTMENT?S COMICS STUDIES MINOR TO PREPARE SHORT GRAPHIC NOVELS EXPLAINING ANTARCTIC BIOGEOGRAPHY, ICEFISH SPECIALTIES, AND THE SCIENCE OF THIS PROJECT AS IT DEVELOPS. AS ANTARCTICA COOLED, MOST SPECIES DISAPPEARED FROM THE CONTINENT?S WATERS, BUT CRYONOTOTHENIOID FISH RADIATED INTO A SPECIES FLOCK. WHAT FACILITATED THIS RADIATION? COYNE?S ?TWO RULES OF SPECIATION? OFFER EXPLANATIONS FOR WHY SPECIES DIVERGE: 1) THE DYSGENIC SEX IN AN INTERSPECIES HYBRID IS THE ONE WITH TWO DIFFERENT SEX CHROMOSOMES (I.E., IN HUMANS, IT WOULD BE XY MALES AND NOT XX FEMALES); AND 2) ?SEX CHROMOSOMES PLAY AN OUTSIZED ROLE IN SPECIATION?. THESE IDEAS PROPEL THE PROJECT?S MAIN HYPOTHESIS: NEW SEX CHROMOSOMES AND NEW SEX DETERMINATION GENES ASSOCIATE WITH CRYONOTOTHENIOID SPECIATION EVENTS. THE MAIN OBJECTIVE OF THE RESEARCH IS TO IDENTIFY NOTOTHENIOID SEX CHROMOSOMES AND CANDIDATE SEX-DETERMINATION GENES IN MANY NOTOTHENIOID SPECIES. THE PROJECT?S FIRST AIM IS TO IDENTIFY ANTARCTIC FISH SEX CHROMOSOMES, ASKING THE QUESTION: DID NEW SEX CHROMOSOMES ACCOMPANY SPECIATION EVENTS? KNOWLEDGE GAPS INCLUDE: WHICH SPECIES HAVE CRYPTIC SEX CHROMOSOMES; WHICH HAVE NEWLY EVOLVED SEX CHROMOSOMES; AND WHICH ARE CHROMOSOMALLY XX/XY OR ZZ/ZW. METHODS INVOLVE POPULATION GENOMICS (RAD-SEQ AND POOL-SEQ) FOR MORE THAN 20 ANTARCTIC CRYONOTOTHENIOIDS. THE PREDICTION IS FREQUENT TURNOVER OF SEX CHROMOSOMES. THE PROJECT?S SECOND AIM IS TO IDENTIFY CANDIDATE ANTARCTIC CRYONOTOTHENIOID SEX-DETERMINATION GENES, ASKING THE QUESTION: DID NEW SEX-DETERMINATION GENES ACCOMPANY ANTARCTIC CRYONOTOTHENIOID SPECIATION EVENTS? A KNOWLEDGE GAP IS THE IDENTITY OF SEX DETERMINATION GENES IN ANY NOTOTHENIOID. PRELIMINARY DATA SHOW THAT THREE SEX-LINKED LOCI ARE IN OR ADJACENT TO THREE DIFFERENT CANDIDATE SEX DETERMINATION GENES: 1) A DUPLICATE OF BMPR1BA IN BLACKFIN ICEFISH; 2) A TANDEM DUPLICATE OF GSDF IN SOUTH GEORGIA ICEFISH; AND 3) A TRANSPOSED DUPLICATE OF GSDF IN STRIPED NOTOTHEN. METHODS INVOLVE ANNOTATING THE GENOMIC NEIGHBORHOODS OF CRYONOTOTHENIOID SEX LINKED LOCI FOR ANOMALIES IN CANDIDATE SEX GENES, SEQUENCING SEX CHROMOSOMES, AND TESTING SEX GENE VARIANTS BY CRISPR MUTAGENESIS IN ZEBRAFISH. THE PREDICTION IS FREQUENT TURNOVER OF SEX DETERMINATION GENES. THE PROJECT?S THIRD AIM IS TO IDENTIFY SEX CHROMOSOMES AND SEX-DETERMINATION GENES IN TEMPERATE NOTOTHENIOIDS. BASALLY DIVERGING TEMPERATE NOTOTHENIOIDS (?BASALS?) LACK IDENTIFIABLE SEX CHROMOSOMES, CONSISTENT WITH TEMPERATURE-CUED SEX DETERMINATION, AND ONE ?BASAL? SPECIES IS A HERMAPHRODITE. THE CONSTANTLY COLD SOUTHERN OCEAN RULES OUT TEMPERATURE, A COMMON SEX DETERMINATION CUE IN MANY TEMPERATE FISH, FAVORING GENETIC SEX DETERMINATION. SOME CRYONOTOTHENIOIDS RE-INVADED TEMPERATE WATERS (?RETURNEES?). KNOWLEDGE GAPS INCLUDE WHETHER BASALS AND RETURNEES HAVE STRONG SEX DETERMINATION GENES. METHODS EMPLOY POOL-SEQ. THE PREDICTION IS THAT GENETIC SEX DETERMINATION IS WEAK IN BASALS AND THAT RETURNEES HAVE THE SAME, BUT WEAKER, SEX-LINKED LOCI AS THEIR ANTARCTIC SISTER CLADE. A PERMANENT EXHIBIT WILL BE ESTABLISHED AT THE EUGENE SCIENCE CENTER MUSEUM TENTATIVELY ENTITLED: THE ANTARCTIC: ITS FISHES AND CLIMATE CHANGE. THOUSANDS OF VISITORS, ESPECIALLY SCHOOL CHILDREN WILL BE EXPOSED, TO THE SCIENCE OF ANTARCTIC ECOSYSTEMS AND THE IMPACTS OF CLIMATE CHANGE. THE RESEARCH TEAM WILL COLLABORATE WITH THE UNIVERSITY?S SCIENCE AND COMICS INITIATIVE TO PRODUCE SHORT GRAPHIC NOVELS EXPLAINING ANTARCTIC BIOGEOGRAPHY, ICEFISH SPECIALTIES, AND THIS PROJECT. THIS AWARD REFLECTS NSF'S STATUTORY MISSION AND HAS BEEN DEEMED WORTHY OF SUPPORT THROUGH EVALUATION USING THE FOUNDATION'S INTELLECTUAL MERIT AND BROADER IMPACTS REVIEW CRITERIA.