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Single Cell Gene Expression of Abnormal and Normal Sperm in the Domestic Cat

Graduate #12
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Audra Huffmeyer - University of California, Los Angeles
Co-Author(s): Robert K. Wayne, UCLA Budhan Pukazenthi and Klaus-Peter Koepfli, Smithsonian Conservation Biology- Center for Species Survival



Carnivore species are at high risk for extinction. This is the result of human expansion, reduction of suitable habitat and dwindling population sizes. As population sizes decrease, species are at risk for increased levels of inbreeding. Increased homozygosity places carnivores at further risk for accumulation of deleterious alleles that are associated reduced fertility. Currently, goals for managing carnivores focus on maintaining suitable habitat and large population sizes. Going forward, maintaining large populations sizes may not be possible. Rather understanding the molecular mechanisms that contribute to abnormal sperm production in carnivores, may contribute to mitigating or reversing abnormal sperm production. Using the domestic cat, as a model, this study identified genes associated with abnormal sperm production as a result of low heterozygosity. We separated abnormal and normal sperm cells from 3 inbred cats and 3 outbred cats. We created two cDNA libraries, one comprised of abnormal sperm cells and the other normal cells, per cat, with the QIAseq FX Single Cell RNA kit. The libraries were then sequenced on an Illumina HiSeq 2500. The sequences were analyzed using RNA-seq, we found genes were differentially expressed between abnormal and normal sperm cells. But, more genes were differentially expressed between abnormal and normal sperm cells in outbred cats. Thus we conclude there is evidence the molecular mechanisms underlying abnormal sperm production in inbred, low heterozygous cats may differ from abnormal sperm production in outbred, heterozygous cats.

Funder Acknowledgement(s): AGEP; NSF GRIP; NSF GRFP

Faculty Advisor: Robert K. Wayne, rwayne@ucla.edu

Role: I separated the cells, completed the library preps, and once the libraries were sequenced I completed the RNA-seq analyses.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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