Figures for Pituitary pathology and gene expression in acromegalic cats

Figure 1

Representative
photomicrographs of growth hormone (A - C) and prolactin (D – F)
immunostaining. A and D are x40 photomicrographs demonstrating specific
immunostaining for somatrophs and lactotrophs, respectively. B and E are
photomicrographs from a control cat C and F are from an acromegalic cat. <br>

Figure 2

Representative images of
SSTR2 immunoreactivity using feline pituitary tissue. A - D represent pituitary
tissue exhibiting SSTR2 immunohistochemistry scores 0, 1, 2 and 3, respectively
using the following criteria:.0 = absent; 1 = cytoplasmic staining; 2 =
membranous staining in less than 50% cells or incomplete membranous staining;
and 3 = circumferential membranous staining in &gt;50% cells. All presented
photomicrographs collected at x100 magnification using Leica DM400 B, Leica
Microsystems Cambridge, UK.<br>

Figure 3

An electropherogram results from
PCR products using multiplex 1 primer sets The blue peaks represent PCR
products from gene specific primers and red peaks represent product size
standards.<br>

Figure 4

All images stained using Silver
stain for reticulin fibres and counter stained using Nuclear Fast Red solution.
A and C; reconstructed stitched pituitary x100 magnification photomicrographs
from two control pituitaries. B and D; x400 magnification photomicrographs from
A and C, respectively. The acinar pattern of reticulin staining is identified
in B and D. This pattern of reticulin staining was demonstrated in all
reticulin staining control pituitaries.<br>

Figure 5

All images stained using Silver
stain for reticulin fibres and counter stained using Nuclear Fast Red solution.
A – D; selected images taken from reconstructed stitched pituitary x100
magnification photomicrographs from four HST pituitaries. A; disrupted
reticulin staining and loss of acinar structure. B; areas of enlarged acini
(blue stars) and areas of loss of acinar structure (blue cross). C; enlarged
acini (blue stars) adjacent to normal sized and small acini. D; loss of acinar
structure in the bottom right of the image (blue stars), and adenomatous tissue
has compressed the normal pituitary tissue resulting in compression of the
acini and a ring of cords of acini giving the impression of a pseudocapsule.<br>

Figure 6

A: Bar charts comparing the
relative gene expression of <i>SSTR1</i>, <i>SSTR2 and</i> <i>SSTR5</i> in pituitary tissue from control (CTRL) and acromegalic
(Acro) cats determine using GeXP multiplex technique. <i>RPL18</i> is the reference gene. Bar height represents mean and error
bars are 95% confidence intervals ** represents <i>P</i> &lt; 0.01 and *** represents <i>P</i>
&lt; 0.001. Dot plot of the individual somatostatin profiles from each of the
19 acromegalic cats.

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Table 1

Clinical data of cats in
the control and acromegalic groups<br>

Table 2

Gene expression data and GH, PRL
and SSTR2 immunohistochemistry scoring of cats in the control and acromegalic
groups.<br>

Table 3

Summary of Spearman rank correlation
gene expression data in the control group and acromegalic groups<br>

Supplementary material 1Nucleotide sequences of primers used in multiplexes 1, 2 and 3.<br>Supplementary material 2Amino acid multiple sequence alignment for human and feline SSTR2<br>Supplementary material 3Amino acid multiple sequence alignment for porcine and feline growth hormone <br>Supplementary material 4Amino acid multiple sequence alignment for porcine and feline prolactin <br><br>