Sherry Cuthbertson
University of Tennessee Health Science Center
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Featured researches published by Sherry Cuthbertson.
Journal of Neuroscience Methods | 1999
Yun Jiao; Z. Sun; Teffy Lee; Francesca Fusco; Toya D.H. Kimble; Christopher A. Meade; Sherry Cuthbertson; Anton Reiner
The masking of antigens by aldehyde-containing fixatives or by paraffin embedding procedures is a problem for immunohistochemical studies. Enzymatic digestion, formic acid treatment, microwave heating and autoclave heating have been used to deal with this problem, with microwave heating-based antigen retrieval having become widely used as the method of choice. Microwave heating, however, has the shortcoming that it is difficult to precisely control the heating temperature and it is difficult to apply this method of heating to free-floating sections without damaging the sections. We describe here a simple, reliable and sensitive antigen retrieval method that uses water-bath heating. By this method, the temperature can be precisely controlled to yield effective antigen retrieval with minimal tissue damage in free-floating or paraffin-embedded slide-mounted sections. We found that the best results were obtained with a 30 min incubation in a 10-50 mM sodium citrate solution (pH 8.5-9.0) preheated to and maintained at 80 degrees C in a water-bath, followed by 30 min incubation in 0.3-3% nonfat dry milk to reduce nonspecfic staining. This method is highly effective for both 40 microm free floating sections, slide-mounted cryostat sections and paraffin-embedded slide-mounted sections, and it works well for tissue from diverse species (human, rat, mouse, pigeon, and zebra finch) and for diverse antigens (e.g. enkephalin, substance P, huntingtin, GluR1, GFAP, and ubiquitin). This method was also found to enhance immunolabeling in glutaraldehyde-fixed tissue that had been prepared for ultrastructural examination, without having a deleterious effect on the ultrastructure.
The Journal of Comparative Neurology | 1997
Sherry Cuthbertson; B. Jackson; C. Toledo; Malinda E.C. Fitzgerald; Yung-Feng Shih; Yuri Zagvazdin; Anton Reiner
Orbital and choroidal blood vessels in mammals are known to receive a parasympathetic innervation from the pterygopalatine ganglion, which appears to utilize vasoactive intestinal polypeptide (VIP) and nitric oxide (NO) to increase choroidal blood flow. The present studies were undertaken to elucidate the anatomical and neurotransmitter organization of the pterygopalatine ganglion input to orbital and choroidal blood vessels in pigeons. Single‐ or double‐label immunohistochemistry were employed on paraformaldehyde‐fixed cryostat sections of the pigeon eye and surrounding orbital tissue to localize 1) VIP+ neurons and fibers; 2) choline acetyltransferase (CHAT)‐containing cholinergic neurons and fibers; 3) axons containing the 3A10 neurofilament‐associated antigen; and 4) neuronal NO synthase (nNOS)‐containing neurons and fibers. NOS+ neurons and fibers were also identified by NADPH‐diaphorase histochemistry in sections and whole‐mount specimens.
Brazilian Journal of Medical and Biological Research | 2003
J.A. Cavani; A. Reiner; Sherry Cuthbertson; Jackson C. Bittencourt; C.A.B. Toledo
The Edinger-Westphal nucleus (EWN) is a central preganglionic parasympathetic cell group that gives rise to cholinergic input to the ciliary ganglion, thereby regulating several neurovegetative ocular functions. Recently, the supposed presence of the neuropeptide urocortin (UCN) has been reported in EWN neurons in rodent brain. The purpose of the present study was to examine the distribution of UCN in avian brain and to investigate by immunohistochemical analysis the possible use of this substance as an EWN marker in a non-mammalian class of vertebrates. Brain tissue of pigeons was incubated with a specific antibody against UCN and the results showed labeling of many small neurons, forming a double wing in the dorsal mesodiencephalic transition area. Their size and shape, however, differed from those of EWN neurons, and they were preferentially located rostral to the EWN. Double-label experiments employing an antibody against the enzyme choline acetyltransferase (ChAT) showed that UCN is not localized to the cholinergic cells of the EWN and confirmed the rostral distributionof UCN never overlapping the ChAT+ EWN cells. Taken together, these results suggest that, at least in pigeons, the UCN+ population does not belong to the traditionally defined EWN.
Vision Research | 1996
Sherry Cuthbertson; Jennifer White; Malinda E.C. Fitzgerald; Yung-Feng Shih; Anton Reiner
The distribution of the ciliary ganglion (CG) innervation to the pigeon choroid was determined immunohistochemically, using antisera against choline acetyltransferase (CHAT) and a neurofilament-related protein (the 3A10 antigen). Single-labeling revealed that the nerve fibers containing these two antigens were similarly distributed in the pigeon choroid, with the superior and temporal quadrants of the eye containing the most fibers. Both types of fibers surrounded and ramified on choroidal blood vessels. Additionally, CHAT+ varicosities were evident among vessels in the choroid and choriocapillaris. Double-label immunofluorescence revealed that CHAT and the 3A10 antigen were almost completely colocalized in choroidal nerve fibers, but absent from CHAT+ varicosities. Substance P-containing and calcitonin gene-related peptide-containing choroidal nerve fibers were poor in 3A10+ labeling. Transection of the postganglionic fibers of the CG reduced CHAT+ and 3A10+ nerve fibers in the choroid to 3-5% of normal abundance, with most of the residual fibers being located in the nasal and inferior quadrants. The present results suggest that the CG in pigeon preferentially influences choroidal blood flow in the superior and temporal parts of the eye, which are involved in high acuity and binocular vision.
Visual Neuroscience | 1999
Sherry Cuthbertson; Yuri Zagvazdin; Toya D.H. Kimble; William J. Lamoreaux; Bryan S. Jackson; Malinda E.C. Fitzgerald; Anton Reiner
The avian ciliary ganglion (CG) controls choroidal blood flow by its choroidal neurons, and pupil constriction and accommodation by its ciliary neurons. It was previously reported that both choroidal and ciliary neurons label positively for NADPH diaphorase (NADPHd), a marker for nitric oxide synthase (NOS). To assess if this labeling is preganglionic or postganglionic and to determine if it is attributable to neuronal NOS (nNOS), we studied pigeon CG using NADPHd histochemistry and nNOS immunohistochemistry (IHC). Short-duration staining times by NADPHd histochemistry yielded intense labeling of structures that appeared to be the cap-like endings on ciliary neurons and the boutonal endings on choroidal neurons that arise from the nucleus of Edinger-Westphal (EW), and light or no postganglionic perikaryal staining. The light postganglionic staining that was observed tended to be localized to ciliary neurons. Consistent with this, NADPHd+ nerve fibers were observed in the postganglionic ciliary nerves but rarely in the postganglionic choroidal nerves. These same staining times yielded robust staining of neurons in the orbital pterygopalatine microganglia network, which are known to be nNOS+. Diffuse staining of CG perikarya was observed with longer staining durations, and this staining tended to mask the preganglionic labeling. Preganglionic NADPHd+ staining in CG with short staining times was blocked by the NOS inhibitors iodonium diphenyl (IDP) and dichlorophenol-indophenol (DPIP), but the diffuse postganglionic staining observed with the longer staining times was not completely blocked. Labeling of CG sections for substance P (SP) by IHC (which labels EW-originating preganglionic endings in CG) and subsequently for NADPHd confirmed that NADPHd was localized to preganglionic endings on CG neurons. Immunohistochemical double labeling for nNOS and SP or enkephalin further confirmed that nNOS is found in boutonal and cap-like endings in the CG. Two studies were then carried out to demonstrate that the nNOS+ preganglionic endings in CG arise from EW. First, NADPHd+ and nNOS+ neurons were observed in EW in pigeons treated with colchicine to enhance perikaryal labeling. Second, NADPHd+ and nNOS+ preganglionic endings were eliminated from CG ipsilateral to an EW lesion. These various results indicate that NOS is present in EW-arising preganglionic endings on choroidal and ciliary neurons in avian CG. NOS also appears to be found in some ciliary neurons, but its presence in choroidal neurons is currently uncertain.
Investigative Ophthalmology & Visual Science | 2003
Sherry Cuthbertson; Mark S. LeDoux; Seth Jones; Julia Jones; Qihong Zhou; Suzhen Gong; Patrick Ryan; Anton Reiner
Experimental Eye Research | 1999
Yung-Feng Shih; Malinda E.C. Fitzgerald; Sherry Cuthbertson; Anton Reiner
Investigative Ophthalmology & Visual Science | 2007
Chunyan Li; Malinda E.C. Fitzgerald; Sherry Cuthbertson; Mark S. LeDoux; Anton Reiner
Investigative Ophthalmology & Visual Science | 2003
Malinda E.C. Fitzgerald; E. Tolley; B. Jackson; Yuri Zagvazdin; Sherry Cuthbertson; William Hodos; Anton Reiner
Investigative Ophthalmology & Visual Science | 2002
Malinda E.C. Fitzgerald; Sv Jones; Sherry Cuthbertson; Anton Reiner