The Measurement of Desoxyribonucleic Acid in Individual Thyroid Cells Prebicusly

Subjected to +°4I Radiation

As this project has progressed, emphasis has shifted
purely physiologic toward the morphologic changes caused by

hat more
From the
I radiatio he Zhe

large bizarre nuclear forms originally found and described at the beg

ine c=

Since ordinary stainizg ne=nods

Suggest the presence of excessive amounts of chromatin material in the Sifarre

te (DT

this project have received increasing attention.

forms that were produced by small doses of 431] plus thiouracil stimlati¢My 2

nmcczar

seemed appropriate to try to measure the amount of DNA actually containedjin such
muclei. Therefore, a histochemical method for measuring desoxyribonucleig acid in
these large nuclei of irradiated thyroids was adapted to this objective.

The method of Pollister for the quantitative microspectr sphotometrig method
of measurement of Feulgen stain within a nucleus was set up in our laborajory with
the suggestions and advice of Dr. Pollister of Columbia University.

Si

the

Feulgen stain combines with DNA in a quantitative fashion, the amount of jtain

contained within a nucleus could be measured by absorbance of transmitted} orzochronm

light having a specific wave length at which there was maximal absorpticn[far this
stain. Rat thyroids were used in these studies. The staining procedure yas carried

out under highly controlled conditions. The irradiated and nonirradiated [tissues
which were to be compared were placed side by side on the same microscopid slide so
that both tissues were exposed to exactly the same steps of preparation afd staining
circumstances. Measurements were made under oil emersion magnification.
With the

use of a calibrated diaphram it was possible to measure the transmission qf l=ght
through a piug of nucleus which was slightly less than the miclear diametqr. The

_

value for absorbance in the plug was then converted to the sphere, which mepresentec
the volume of the micleus.

The mean diameter of each nucleus was determirpd DSy

direct micrometer measurements. DNA was recorded in arbitrary units.
Tha vate for
a normal resting micleus was determined and standardized against rat li erp ceiis. A
value approximately equal to the diploid value in the rat cell was obtaing#i /&=<m
measurement on the liver.

When thiouracil which is a powerful stimulus to hyperplasia of the thyrcid ws

given to an animal, the diameters of the nuclei of normal thyroid cells :npre=sed.

The DNA also increased toward values which represented two times the dipidid w=lue, 2t
which point the cell normally is expected to divide.

Rats were given 10, 30, or 100 pic of radioiodine after a temporary ppriod cf codideficiency to inhance the uptake of the 131], Thereafter a diet containing a moral
amount of iodine was resumed. One half of each group of animals was then Bivem thicrec
in their drinking water. Same of each group of animals were sacrificed atj16 weexs,
the remaining at 26 weeks.
None of the doses of 1311 used in this series of experiments was sufficr=ntly
great to cause architectural distortion of the thyroid. However, the largRst Sioses

were sufficient to interfere with the expected increase in thyroid weight
usually produced in normal thyroids by thiouracil.

The lesser doses of

fhat is

ragiat=on

Select target paragraph3