THE EFFECT OF FETAL AND

EARLY POSTNATAL IRON

DEFICIENCY ON IRON
METABOLISM IN ADULT
RATS

Christina Sanabria

Jinyan Fu,
Anqiang Yang,
Yajing Ma, Min
Lui, Liciui
Zhang, Yong
Wang, Liegang
Liu

INTRODUCTION
Pregnant rats were administered low iron diet (LD)
Study explored iron metabolism in the of fspring of pregnant
Windsor rats during adolescence and adulthood.

Investigation of early fetal and infant iron deficiency on iron

metabolism later in adulthood.

KEY TERMS
LD: low iron diet during pregnancy (1 st stage)
CD: control-iron diet during pregnancy(1 st stage)
TIBC- Total iron binding capacity
TF: transferrin
TfR- transferrin receptor

DMT1- Divalent metal transporter 1

EPO- erythropoietin

BACKGROUND

BACKGROUND

STAGE 1
Three stages represented in the study.
Stage 1 focused on 30 young adult females aged 6 -7weeks
Fed standard chow containing 250mg/ iron/kg for one week
before being divided into 2 groups after mating
LD group was fed a diet containing 7.4mg/iron/kg

CD group was fed a diet containing 274mg/iron/kg

STAGE 2
Control diet initiated to both LD, and CD pups for a total of 10
weeks. (3-13weeks)
Ensured both groups attained the same level of iron nutrition
at an early stage of life.
Iron balancing period.
8 of the 15 rats in each group (LD -LD & LD-CD) were
sacrificed at the end of stage 2 by decapitation.

STAGE 3
13-29 weeks of a LD diet
At the end of stage 3, all groups were deprived of food for a
total of 12 hours before the remaining rats were sacrificed

Venous blood samples, encephalon and livers collected

METHODS
Binding Assay
ELISA Assay
Radioimmunoassay
Real time PCr
Immunohistochemical (DMT-1) Analysis
Immunofluorescence

BINDING ASSAY
Serum iron and TIBC analyzed
Detection method, usually radioactive
Radioactive iron is filtered, while bound iron is trapped and
analyzed
*Fe (ligand)+Transferrin (Free)Fe-Transferrin(Bound)

Measures how strong binding is: Bound/Free Fe

ELISA ASSAY

Enzyme linked immunosorbent assay

Measured Hb concentrations

Uses enzyme as a second antibody that can be stained

Results in an antibody bound to the enzyme

more color = more protein (TF)

RADIOIMMUNOASSAY
Labeled radioactive ferritin antigen (tracer) is used
and held constant
Unlabeled (nonradioactive) Ferritin added, competing
with Ferritin* for receptor binding sites
Increased amount of unlabeled Ferritin displaces
Ferritin*, resulting in free Ferritin*
Determines how much ferritin is actually present

IMMUNOFLUORESCENT
Kidney tissues
incubated
A fluorescent tag is
used in place of
radioactive tag
TF binding to TfR in
order to enter tissues
See (Fig. 13.5)
Found primarily in
mesangial tissues of LD
rats
Mesangial area of kidneys

REAL PCR

Analyzed the mRNA of kidney EPO

EPO- induces red blood cell synthesis in cells

IMMUNOHISTOCHEMICAL

Tissues obtained and incubated with primary antibody

against DMT1 or ferroportin
See Fig 13.3

Purpose: look for an increased presence of transporters in

cells

RESULTS

Ef fects of Iron Deficiency During Pregnancy on Birthrate

Live births of LD dams 84.7%
Live births of CD dams 95.4%

Ef fects of Iron Deficiency during pregnancy on Infant growth

Stage 1: LD offspring had significantly lower Hb levels in the liver

and serum and higher RET when compared to CD offspring

Characteristic of iron deficiency anemia

RESULTS
Influence of Iron Balancing on Growth and Iron Metabolism

Iron balancing period

Rapid weight gain in both CD and LD groups
Both male and female LD rats exceeded mean weight gain when
compared to CD

Ef fect of Low -Iron Treatment in Adulthood on Hematological

Parameters and on Expression of Duodenal DMT -1 , Renal
TfR, and EPO
Significant increased TIBC, TF, serum EPO,
Expressed increased DMT-1 and renal TfR, as well as higher ferritin
levels

DISCUSSION

1 st stage iron deficiency significantly influenced

growth rate of newborn rats
survival rates of LD and CD offspring
Inadequate iron storage

Long term effects on offspring (Stage 2 & 3)

Suggest maternal iron deficiency may have a positive effect on iron
metabolism in offsprings adolescence and adulthood .

CONCLUSION

Further studies need to be conducted, due to unexpected

results of LD rats increased iron absorption capabilities as
adults even after iron deficiency in their early life.