Publication: Melatonin-like immunoreactivity in the pineal
gland of the cow: an immunohistochemical study
Authors
Carvajal, J.C. ; Gómez Esteban, M.B. ; Carbajo, S. ; Muñoz Barragan, L.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
With a view to checking the presence of
melatonin in the pineal gland of the cow, in the present
work we used six adult animals, ranging in age from one
to six years, which were sacrificed at dawn. Sections of
6 µm thickness of Bouin-fixed and paraffin-embedded
pineal glands were incubated in an anti-melatonin serum,
which was provided by the Institute for Molecular and
Cellular Recognition, Gunma University, Maebshi,
Japan. After incubation and successive washings in PBS,
some of the sections were treated with the avidin-biotinperoxidase
complex (ABC) technique using antisera
from Sigma, and developed with the method of Graham
and Karnovsky (which employs 3,3’-diaminobenzidine
and H2O2 as developer). Other sections were incubated
in a goat-anti-rabbit IgG (H+L) bound to fluorochrome
Cy5 for immunofluorescence studies. An intense
reaction for melatonin was observed in the cytoplasm
but not in the nucleus of melatonin secreting
pinealocytes located in peripheral and intermediate
zones of the pineal gland. Immunoabsorption of the
antimelatonin primary antibody with melatonin at a
dilution of 10 mM per 0.1 ml of serum prevented the
reaction, as happened when any of the antisera used in
the procedure were used. Immunoabsorption of antimelatonin
serum with different amounts of bovine
albumin (ranging between 1/5 to 1/50) failed to inhibit
the immunoreactivity. When a bovine anti-albumin
antibody was employed, working with the above
methods, no immunoreaction was detected. Our data
suggest that the pinealocytes of cows sacrificed at dawn
contain immunoreactive melatonin.
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