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.

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 >50% cells. All presented
photomicrographs collected at x100 magnification using Leica DM400 B, Leica
Microsystems Cambridge, UK.

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.

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.

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.

Figure 6

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

Table 1

Clinical data of cats in
the control and acromegalic groups

Table 2

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

Table 3

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

Supplementary material 1
Nucleotide sequences of primers used in multiplexes 1, 2 and 3.

Supplementary material 2
Amino acid multiple sequence alignment for human and feline SSTR2

Supplementary material 3
Amino acid multiple sequence alignment for porcine and feline growth hormone

Supplementary material 4
Amino acid multiple sequence alignment for porcine and feline prolactin