Presence of nerves and their receptors in mouse and human conjunctival goblet cells.
OBJECTIVE
To determine whether neural pathways for controlling goblet cell secretion are present in mouse and human conjunctiva.


METHODS
Mouse conjunctiva was homogenized and subjected to electrophoresis and Western blotting to detect PGP 9.5 (indicates nerves), muscarinic receptor subtypes (indicates parasympathetic pathway), and adrenergic receptors (indicates sympathetic pathway). Mouse eyes and human conjunctival tissue were analyzed by immunofluorescence microscopy. Antibodies to vasoactive intestinal peptide (VIP), tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and muscarinic and alpha(1)- and beta-adrenergic receptor subtypes were used.


RESULTS
Western blot demonstrated PGP 9.5, M(1), M(2), and M(3) muscarinic receptors and alpha(1A)-, beta(1)-, beta(2)-, and beta(3)-adrenergic receptors in mouse conjunctiva. Immunoreactivity for VIP, TH, and DBH was found adjacent to mouse and human goblet cells. M(1) and M(2) muscarinic receptors were identified throughout mouse conjunctiva, but M(3) receptor was predominantly on goblet cells. All three muscarinic receptor subtypes were detected on goblet cells in human conjunctiva. alpha(1A)-Adrenergic receptors were found on epithelial cells and on goblet cells in mouse and human conjunctiva. beta(1)- and beta(2)-Adrenergic receptors were found on both epithelial and goblet cells in mouse conjunctiva, but not on human conjunctival cells. beta(3)-Adrenergic receptors were found on both epithelial and goblet cells in human conjunctiva but not on mouse conjunctival cells.


CONCLUSIONS
The following conclusions were drawn: parasympathetic nerves and M(1), M(2), and M(3) muscarinic receptors, as well as sympathetic nerves are present on mouse and human goblet cells. The adrenergic receptors beta(1) and beta(2,) but not alpha(1A) and beta(3) are present on mouse conjunctival goblet cells, whereas alpha(1A) and beta(3,) but not beta(1) and beta(2) are present on human conjunctival goblet cells, suggesting that these nerves and receptors could activate goblet cell secretion in mouse and humans.
Diebold
Y
Y
Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Ríos
J D
JD
Hodges
R R
RR
Rawe
I
I
Dartt
D A
DA
D000368:Aged; D000818:Animals; D015153:Blotting, Western; D003228:Conjunctiva / Q000294:innervation* / Q000378:metabolism; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D020397:Goblet Cells / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010275:Parasympathetic Nervous System / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011941:Receptors, Adrenergic / Q000378:metabolism*; D011976:Receptors, Muscarinic / Q000378:metabolism*; D013564:Sympathetic Nervous System / Q000378:metabolism*; D013869:Thiolester Hydrolases / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
D011941:Receptors, Adrenergic; D011976:Receptors, Muscarinic; D014660:Vasoactive Intestinal Peptide; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase