Fig 1.
Mental gland of K. koreana (A, B) and D. ocoee (C). The mental gland is an aggregate of simple exocrine glands. Secretory cells line the periphery of the gland and contain a granular product. The cytosol of K. koreana secretory cells are positive for SPF immunoreactivity, indicated by a dark red colored product (A). The antibodies were made against SPF proteins from D.ocoee mental glands. Desmognathus ocoee mental gland tissue had SPF immunoreactivity as expected (C). The mental gland is PAS positive as well, indicated by the magenta reation product in the cytosol of the secretory cells and for secretory products in the gland lumen (B). Results were consistent between inidividuals (n = 3 for each species). Scale bars are 100 μm. N = nucleus; C = cytosol; Lu = lumen; Sd = secretory duct; Ep = epidermis.
Fig 2.
Negative controls for K. koreana caudal courtship glands (A), and mental gland (B), without SPF primary antibody. Negative controls, treatments without the primary antibody, were used to assess background and non-specific staining. No dark red colored product is visible in these controls indicating the absence of non-specific staining from the secondary antibody. Methyl green was used as a nuclear stain to identify cells. This negative control treatment was used to assess background and non-specific staining. The arrows point to the secretory cells within the caudal courtship (A) and within an individual gland of the mental gland. Scale bars are 100 μm, n = 3. Ccg = caudal courtship gland, Dgg = dorsal granular gland, Mg = one individual gland within the mental gland.
Fig 3.
Caudal courtship and dorsal granular glands in K. koreana tail base tissue.
Secretory products in dorsal granular glands are positive for SPF immunoreactivity, indicated by a dark red colored product (A). In contrast, the secretory cells of caudal courtship glands are negative for SPF immunoractivity, when tested with antibodies made against SPF proteins from the D.ocoee mental gland (A). Interestingly, the caudal courtship glands are positive for PAS (magenta in color (B)), while the dorsal granular glands are negative (B). Caudal courtship glands are similar in color to mental gland treated with PAS (Fig 1B). Results were consistent among individuals (n = 3). Scale bars are 100 μm. Ccg = caudal courtship gland, Dgg = dorsal granular gland.