Carbohydrate metabolism
of wool follicles
"Much of the
research into nutrition and wool growth has centred on the supply
and utilisation of amino acids by the follicle, but very little
research has examined the means by which follicles obtain and use
energy substrates. My work is the first to look at energy metabolism
in detail in wool follicles.
The first experiment I
conducted was aimed at determining whether or not the glycogen
levels in wool follicles are altered by nutrition, and whether
glycogen content is related to wool growth rate. To examine this, I
placed groups of sheep on restricted plans of nutrition and then
allowed them to rapidly gain weight. The restricted nutrition groups
lost approximately 20% of their original liveweight over the 12
weeks of declining nutrition and rapidly regained weight during the
six-week recovery period. Wool production, fibre diameter, staple
length growth rate, muscle glycogen concentration and follicle bulb
division rate all reflected the pattern of liveweight loss. Skin
glycogen concentration was responsive to nutrition and there was a
strong correlation between skin glycogen concentration and wool
production.
In this experiment I
experienced problems in measuring glycogen levels in biopsied skin
tissue. The variation in glycogen concentration between sample times
exhibited a regular pattern, most likely an artifact of sampling
rather than a biological phenomenon. There are two factors that may
have contributed to the weekly variation in glycogen concentration
evident in Experiment 1:
-
inconsistent
extraction of glycogen from the skin tissue; and
-
variation in
glycogen concentration at different sampling sites.
After a good deal of
experimentation, I tracked the problem down to poor repeatability of
extraction of the glycogen from the biopsy tissue, and found that a
potassium hydroxide extraction method worked well.
It was established during
Experiment 1 that the skin glycogen concentration responds to
nutritional manipulation. The objectives of the next experiment were
to examine the dynamics of skin glycogen in genetically high and low
staple strength animals and to demonstrate that glucose transporter
type 1 (GLUT1) expression changes in parallel to glycogen dynamics
in ovine skin. The nutritional simulation of feed availability at
the break of season was repeated in this experiment. Liveweight,
wool production, fibre diameter, staple length growth rate and
staple strength demonstrated similar trends to Experiment 1. These
parameters were significantly affected by nutrition, but not by
genetic staple strength line.
Another experiment was then
conducted to localise GLUT1 protein distribution and to examine
expression of GLUT1 in ovine skin in response to a short-term oral
glucose load. The hypothesis was that an oral glucose load will
up-regulate GLUT1 expression in the wool follicle. The experiment
was conducted for five days. Four treatments were used to test the
hypothesis:
-
control (water)
-
drench at 8-hour
intervals
-
glucose drench
at 8-hour intervals, glucose drench at 12-hour intervals
-
lupin fed at
8-hour intervals.
Glycogen content of the skin
did not differ significantly between treatments. I have had no
success in detecting GLUT1 protein in sheep skin using an
immunological approach, due to problems with the antibody. We have
decided to switch to a molecular approach to detect Glut-1 mRNA."
The final set of experiments
investigated the medium and short-term manipulation of glycogen in
the skin. Glucose was infused intravenously in an attempt to
increase the amount of glycogen stored in the skin. These
experiments will be finalised in the near future.
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