MOLECULAR DIFFERENCES
RESULTING FROM LARVAL
GROWTH CONDITIONS IN
AEDES AEGYPTI
DAVID PRICE
DISEASE VECTOR MOLECULAR BIOLOGY LAB
NMSU
PI 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 Meal
NBF- 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Price et al, unpublished data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Price et al, unpublished data,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?
Reproduction Transcripts
vitellogenin-A1 precursor
vitellogenin-C
vitellogenin-B
SPBM
LPBM
Vitellogenic carboxypeptidase Precursor
cathepsin b
0
4,000,000
8,000,000
Read Count
12,000,000
Immune and Most Changed Transcripts
Cecropin
Defensin
Relish
Cactus
Trypsin 3A1 Precursor
SNBF
LNBF
serine-type enodpeptidase
Niemann-Pick Type C-2, putative
trypsin
hypothetical
1
10
100
1,000
Read Count (Log10 Scale)
10,000
100,000
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