Molecular Medical Genetics 16-13

M13 pUC18
A sequencing ladder

CAG Repeats: 16/16 24/41 20/42 15/18 16/19 I'¢.: ~~.,

'176' "'7

B CAG Repeats:

~=Il- .._' '"--' ., . '":-,... , 16/19

'~~~.
.... --------_r- ,00
16/19

21/39
o "
.... ft
o_·,_IV~~
.~o ~
.....v,...>r-,---- ----z·oV- .~-
16/28

~~,T--------~~Q;~----__zl,~ 21/39
,.., ......

o~g~a----~"-":Ao-~--- -z,~ , ...., 26/40

"'"
o- - 1tIr--- -JJ.Ir,----~~~---__z,Gtr_, ..... 26/40
"'" Size
o---4,b--- - - - - - - - -- - - - - - -i" L "'" markets

Fig. 5. Molecular genetic testing of HD. (A) An autoradiograph of radiolabeled PCR products that encompass the HD CAG repeat
following acrylamide gel electrophoresis. The number of individual CAG repeats is calculated by comparison to an M13
sequencing ladder and are noted below the gel. (B) Acrylamide electrophoresis of fluorescently labeled PCR products that
encompass the HD CAG repeat detected by an automated sequencer. The number of individual CAG repeats is calculated by
compari son with standard size markers and are noted to the right of the graphical output. (Images courtesy of M Galvez, P Scott,
and D Rosenblatt, McGill University Health Centre, Division of Medical Genetics, Montreal , Canada .)

• Variable findings include pyramidal and extrapyramidal ATXN3

signs, nystagmus, amyotrophy fasciculations, and
sensory loss
Prevalence
• Accounts for approximately 20% of all SCA
Diagnosis
• Very difficult to distinguish from other hereditary ataxias ;
requires molecular genetic testing to detect an abnormal
CAG trinucleotide repeat expansion in the coding region
of the ataxin 3 (ATXN3) gene (Figure 4 and Table 4)
• Normal alleles have up to 47 repeats
• Unlike SCA2 , phenotypically normal individuals can
have intermediate allele s that contain 48-51 repeats;
can result in pathologic expansion in subsequent
generations
• Affected individuals have at least one allele with 53-86
CAG repeats
• Increase in disease severity has been observed in
individuals homozygous for expanded ATXN3

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