MOLECULAR DIFFERENCES RESULTING FROM LARVAL GROWTH CONDITIONS IN AEDES AEGYPTI

DAVID PRICEDISEASE VECTOR MOLECULAR BIOLOGY LABNMSUPI IMMO HANSEN

Background

Aedes aegypti is an important disease vector

Relatively safe model for malaria Plasmodium gallinaceum

Existing Reference Genome

Aedes/Dengue Situation

http://www.healthmap.org/dengue/index.php

Small vs Large Mosquitoes

Larval conditions affect adult insect Crowding, food availability ->Changes in adult size, time to reach

adulthood, immunity, reproductive success, energy stores, life span, fecundity on first blood meal, vectorial capacity

Fat Body

Bloodfeeding

Purdue Medical Entomology

PBM- Post Blood MealNBF- Not Blood Fed

Hypothesis

Significant metabolomic, transcriptional and metabolic differences exist in the fat body between small and large mosquitoes, before and after blood meal.

Methods

Metabolic CO2 production

Metabolomic GC/MS, LC/MS, LC/MS/MS analysis

Transcriptomic RNA-Seq approach using Illumina sequencing

Metabolomics

Questions: Map metabolic pathway up/down regulation

after blood meal

Price et al, unpublished data

Metabolic

Questions: Do small and large mosquitoes burn energy

at the same rate? Are the substrates being used for energy

the same?

In conjunction with other work: Is there a difference in how far energy stores

go?

Metabolic

Price et al, unpublished data, Sable Systems

Transcriptomic

Questions: Determine changes in gene expression

between mosquitoes raised under different nutrient conditions

What patterns of expression are present?

cathepsin b

Vitellogenic carboxypeptidase Precursor

vitellogenin-B

vitellogenin-C

vitellogenin-A1 precursor

0 5,000,000 10,000,000 15,000,000

Reproduction Transcripts

SPBMLPBM

Read Count

hypothetical

trypsin

Niemann-Pick Type C-2, putative

serine-type enodpeptidase

Trypsin 3A1 Precursor

Cactus

Relish

Defensin

Cecropin

1 10 100 1,000 10,000 100,000

Immune and Most Changed Transcripts

SNBFLNBF

Read Count (Log10 Scale)

Conclusions

Used metabolomic, metabolic, and transcriptional descriptive work to identify differences between small and large mosquitoes; pre and post bloodmeal.

Immune regulation is curious Results of small vs large vectorial capacity

experiments have been conflicting Signs of autophagy No apparent metabolic difference

Hansen Lab/Collaborators

Alexander Ulanov – University of Chicago, Urbana

Wayne Van Voorhies - NMSU molecular Biology

NIAID/NIH