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. 2002 Jul;201(1):41-52.
doi: 10.1046/j.1469-7580.2002.00070.x.

Parathyroid hormone-related protein (PTHrP) production sites in elasmobranchs

M K Trivett  1 , T I Walker, D L Macmillan, J G Clement, T J Martin, J A Danks
Affiliations

Parathyroid hormone-related protein (PTHrP) production sites in elasmobranchs

M K Trivett et al. J Anat. 2002 Jul.

Abstract

This study describes the distribution of parathyroid hormone-related protein (PTHrP) antigen and its mRNA in seven species of cartilaginous fish from six elasmobranch families. Antigen was detected using antibodies to synthetic human PTHrP and the mRNA with a riboprobe to human PTHrP gene sequence. The distribution pattern of PTHrP in the cartilaginous fish studied, reflected that observed in mammals but PTHrP further occurs in some sites unique to cartilaginous fish. Of particular note was the demonstration of PTHrP in the shark skeleton, which although considered not to contain bone, may form by a process similar to that forming the early stages of mammalian endochondral bone. The distribution of PTHrP in the elasmobranch skeleton resembled the distribution of PTHrP in the developing mammalian skeleton. Differences in the staining pattern between antisera to N-terminal PTHrP and mid-molecule PTHrP in the brain and pituitary suggested that the PTHrP molecule might be post-translationally processed in these tissues. The successful use of antibodies and a probe to human PTHrP in tissues from the early vertebrates examined in this study suggests that the PTHrP molecule is conserved from elasmobranchs to humans.

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Figures

Fig. 1
Fig. 1
(a) Gummy shark skin immunostained with PTHrP(1–14) antiserum. PTHrP antigen is seen in the epidermis (E), immediately adjacent to the spine (S) of the mature dermal denticle, with weaker staining throughout the remainder of the epidermis, but not in the dermis (D). Pigment (black) is visible. (×110). (b) In situ hybridization of gummy shark skin showing PTHrP mRNA in the epithelium (E) and pulp cavity (P) of developing denticles. Weak signal for the PTHrP mRNA occurs throughout the remainder of the epidermis but not in the dermis (D) (×110). (c) Section of shark gill immunostained with N-terminal PTHrP antiserum showing PTHrP antigen in the crypt or ‘chloride’ cells (Cr), epithelium of the interlamellar (I) and secondary lamellar (S) epithelium. The PTHrP antigen is not seen in the gill ray (R). (×110). (d) Tubules (T) in angel shark rectal gland are tightly packed and interspersed with connective tissue (CT). PTHrP immunoreactivity is seen in tubular epithelial cells and in scattered blood cells (B) but not in the connective tissue. (×110). (e) PTHrP immunohistochemistry showing PTHrP antigen in proximal (P) and distal (D) tubules of the kidney. Antigen is absent from the neck segments (N) and glomeruli (G). (×55). (f) Non-immune control of 1e, showing absence of PTHrP staining (lettering as in 1e, ×55).
Fig. 2
Fig. 2
(a) Von Kossa staining shows the sites of calcification (brown) in the gummy shark vertebrae. The outer notochordal sheath (O) is calcified but the inner sheath (I) is not. Vaculoated notochordal cells are not visible. Calcified cartilage (CC) occurs at the periphery of the vertebra and in association with the outer notochordal sheath. Uncalcified cartilage appears orange to red. Associated muscle (M) is visible. (×14). (b) PTHrP immunoreactivity is seen in the cytoplasm of chondrocytes (arrow) at the edge of the calcification front that lies between the calcified cartilage (CC) and perichondrium (P). Chondrocytes within the uncalcified cartilage (C) stain for PTHrP antigen, as does skeletal muscle (M). (×137.5). (c) Staining for the PTHrP antigen in the developing tooth is seen in the odontoblasts (O), and the epithelial cells (E) overlying and adjacent to the developing tooth. Some scattered cells in the pulp cavity (P) stain, but the dentine (D) does not. (×110). (d) An adjacent section stained with antiserum to PTH (1–34) shows no immunoreactivity (lettering as in 2c, ×110).
Fig. 3
Fig. 3
(a) Immunohistochemistry shows staining in neurones (N) in the tectum that border the ventricle (V). The ventricle is lined by a layer of ependymal cells (Ep). The surrounding tissue is negative. (×110). (b) No staining of the neurones, ependymal cells lining the ventricle is seen with the PTH antiserum (lettering as in 3a, ×110). (c) Immunohistochemistry shows PTHrP antigen in epithelial cells (E) of the choroid plexus and in ependymal cells (Ep). Only scattered cells in the capillaries (C) stain for the PTHrP antigen. (×55). (d) In the spinal cord, hybridization signal for the PTHrP mRNA is seen in the grey matter (G) and, to a lesser extent, in ependymal cells lining the central canal (C). No hybridization is seen in white matter (W). (×55). (e) Angel shark pituitary incubated with mid-molecule PTHrP antiserum shows staining at the edge of the neurointermediate lobe (NL) and the stalk (S) connecting the pituitary to the diencephalon (D). Part of the saccus vasculosus (SV) is visible. (×55). (f) An adjacent section incubated with N-terminal PTHrP antiserum shows staining in the endocrine cells of the neurointermediate lobe but not in the stalk or diencephalon. Part of the saccus vasculosus is visible (lettering as in 3e, ×55
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