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SnRNA-seq of murine tarsal plates

ABSTRACT: The eyelid is a complex environment containing, containing epidermal, hair follicles, and conjunctival cells closely adjacent to the Meibomian glands (MGs), making it challenging to isolate pure MG cells or to identify MG contributions in bulk RNA-seq analyses. To analyze transcription in MG subpopulations, and to overcome difficulties in isolating cytoplasmic mRNAs from lipid-rich acinar cells, we utilized snRNA-seq which allows for identification of cell sub-populations within a complex mix.

ORGANISM(S): Mus musculus

PROVIDER: GSE274498 | GEO | 2025/01/21

REPOSITORIES: GEO

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Project description:Native host plant insect resistance in the maize inbred line Mp708 was developed by traditional plant breeding. Resistant Mp708 thwarts feeding by fall armyworm (Spodoptera frugiperda [J.E. Smith]; Lepidoptera: Noctuidae), numerous other lepidopteran pests, and the coleopteran western corn rootworm. This broad resistance makes it an excellent model for studying native host plant resistance mechanisms. In response to caterpillar feeding, Mp708 rapidly mobilizes Mir1-CP, a unique cysteine protease that appears to translocate from roots to the maize midwhorl where it accumulates. This accumulation correlates with a significant reduction in caterpillar growth resulting from diminished food utilization. In addition, the peritrophic membrane (PM) that surrounds the food bolus in the mudgut (MG) is severely damaged in caterpillars fed on sweet corn callus transformed to express the gene encoding Mir1-CP or on midwhorl tissue from resistant Mp708 maize. Functions of the PM include assisting digestion and protecting the epithelium of the caterpillar MG from physical and chemical damage. Consequently, the reduced growth of caterpillars that feed on Mp708 is probably due to the action of Mir1-CP on PM physiology. In fact, previous in vitro studies indicated that Mir1-CP was capable of permeabilizing the PM. The present study used both targeted (qRT-PCR) and global (mRNA-seq) transcriptome analyses to explore the effect of eating Mir1-CP expressing Mp708 maize on abundance of transcripts in the MG of fall armyworm larvae in comparison to MGs from larvae fed on susceptible Tx601 maize that does not express Mir1-CP. Expression of genes encoding proteins involved in PM production is upregulated in MGs from fall armyworm fed on Mp708. Also, several digestive enzymes (endopeptidases, aminopeptidases, lipases, amylase) were more highly expressed in MGs from larvae fed on Mp708 than MGs from larvae fed on Tx601. Impaired growth of larvae fed on Mp708 probably results from metabolic costs associated with higher production of PM constituents and digestive enzymes in a compensatory attempt to maintain MG function.

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SnRNA-seq of murine tarsal plates

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