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A preliminary analysis of the southern white rhinoceros granulosa cell transcriptome (Ceratotherium simum simum)

ABSTRACT: In vivo-collected granulosa cells (GC) from the southern white rhinoceros (SWR) provide a non-invasive assessment of the developmental status of oocytes prior to in vitro culture, which will aid in the development of assisted reproductive technologies (ART). Our study aimed to investigate gene expression in SWR granulosa cells (GC), collected in vivo and gain preliminary insight into the transcriptional activity occurring within the cells during various stages of oocyte development. It was hypothesized there would be similarities between the SWR GC transcriptome and cattle and humans, two species for which well-annotated genomes are available and ART are commonly used. GC were collected from SWR following ovum pickup (OPU) and pooled from all aspirated follicles. Total RNA was isolated, libraries prepared and sequencing performed using an Illumina NextSeq 500. Reads were aligned and annotated to CerSimCot1.0. Databases for cattle and human were acquired for comparison. This study identified 37,407 transcripts present in GC of SWR. It was determined that cattle and human transcriptomes are valuable resources with a homology of 45% with the SWR. In conclusion, these data provide preliminary, novel insights into the transcriptional activity of GC in the SWR that can be used to enhance ART in this species.

ORGANISM(S): Ceratotherium simum simum

PROVIDER: GSE216035 | GEO | 2023/05/11

REPOSITORIES: GEO

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Project description:Investigation of cellular responses to thermal heat stress can lead to identifying novel targets of intervention to tackle the increasing fertility decline due to climate change-associated seasonal heat stress. We aimed to study the response of granulosa cells to seasonal thermal stress. Dry beef cows (n=11) were kept together during the study and subjected to synchronization and stimulation for follicular dynamics. Ovum pick-up (OPU) was conducted on all animals in the winter and summer. Once OPU-fluid settled, cumulus-oocyte-complexes (COCs) and granulosa cells were isolated from the OPU fluid and stored in a -80 freezer after snap freezing. In addition, blood samples and rectal temperature data were collected on each OPU. Environmental data were collected daily for three weeks before each OPU using the Florida Automated Weather Network. Data were analyzed using SAS. Data are presented as mean ± standard error of the mean. RNA was extracted from five pools of granulosa cells followed by library preparation and sequencing (NovaSeq; Illumina). As expected, environmental conditions were contrasting: average air temperature (11.5°C vs. 27.5°C), average max air temperature (16.9°C vs. 33.7°C), relative humidity (83.5% vs. 82.3%), and temperature-humidity index (53.39 vs. 79.16) for winter and summer, respectively. The average rectal temperature was higher (P = 0.03) in summer (39.2 ± 0.2°C) than in winter (38.8 ± 0.2°C). Of the total differentially expressed genes, granulosa cells during summer showed up and downregulation of 1083 and 1126 transcripts, respectively compared to the winter season (Fold Change ≤ -2 or ≥ 2 and FDR p-value ≤ 0.05). Many of the downregulated transcripts were found to be involved in the inhibition of immunological pathways such as activation of lymphatic system cells and activation of lymphocytes. In conclusion, exposure of cows to thermal stress can induce significant alteration in the transcriptome of granulosa cells which may have a negative impact on follicular physiology.

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A preliminary analysis of the southern white rhinoceros granulosa cell transcriptome (Ceratotherium simum simum)

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