Immunohistochemical procedures for the demonstration of peptide- and tyrosine hydroxylase-containing nerve fibers in cryostat sections of unfixed rapidly frozen tissue stored for long periods of time

Forsgren, Sture; Söderberg, Lena

doi:10.1007/bf00492471

Cited by 54 publications

(12 citation statements)

References 20 publications

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“…However, Chen et al (1998) considered that the sympathetic neurotransmission is the principal source of inhibitory neural input to the gallbladder smooth muscles, causing decreased gallbladder tone, and may contribute to the filling of the organ. In the current study TH positive nerve fibres were observed as single fibres, nerve fibres forming perivascular plexuses and nerve fibres grouped into nerve fascicles, which correlates with the results of Forsgren & Söderberg (1987) regarding the distribution of the mentioned nerves in the guinea pig and bovine heart. Our findings indicate that the nerve supply of gallbladder in swine was similar to those of guinea pig gallbladder, described by Cai & Gabella (1983).…”

Section: Discussionsupporting

confidence: 90%

Tyrosine hydroxylase positive nerves and mast cells in the porcine gallbladder

Stefanov¹,

Vodenicharov²,

Tsandev³

et al. 2017

BJVM

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The aim of this study was to detect the localisation of tyrosine hydroxylase (TH) positive nerve fibres (THN) and distribution of tyrosine hydroxylase positive mast cells (THMC) in the wall of porcine gallbladder. THN were observed as single fibres, nerve fibres forming perivascular plexuses and nerve fibres grouped within the nerve fascicles. In the gallbladder`s fundus, body and neck, the TH + fibres formed mucosal, muscular and serosal nonganglionated nerve plexuses. Toluidine blue (TB) staining was used to confirm that the TH positive cells were mast cells. The number of THMC in the propria of gallbladder`s fundus, body and neck was significantly higher than in the muscular and serosal layers in both genders. The number of mast cells in the musculature was higher than in the serosa. The density and location of the THMC were similar to the TB positive (with gamma metachromasia) mast cells in both males and females, and statistically significant difference was not established. In conclusion, original data concerning the existence and localisation of catecholaminergic nerves and THMC distribution in the porcine gallbladder's wall are presented. The results could contribute to the body of knowledge of functional communication between autonomic nerves and mast cells in the gallbladder.

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Section: Discussionsupporting

confidence: 90%

Tyrosine hydroxylase positive nerves and mast cells in the porcine gallbladder

Stefanov¹,

Vodenicharov²,

Tsandev³

et al. 2017

BJVM

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“…The slides were then rinsed with Tris-HCl-buffered saline (TBS, 0.05 M , pH 7.6) and fixed in acetone at –20°C for 20 min. Double staining for surface antigen CD3 and cytoplasmic antigen SP of IEL and LPL was performed by the immunofluorescence technique [38]. In brief, the bound cells were incubated with 0.5% bovine serum albumin (BSA) in TBS for 30 min to block the nonspecific staining, and then treated with rat anti-mouse CD3 mAb (50 µg/ml, KT3, MCA 500G, Serotec) in a moist chamber at 37°C for 1 h. After washing, the slides were incubated successively with 0.5% BSA for 15 min and fluorescein isothiocyanate (FITC)-labeled goat anti-rat (F(ab′) 2 fragment (30 µg/ml, Jackson ImmunoResearch Laboratories, West Grove, Pa., USA) at 37°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Both Substance P and Its Receptor Are Expressed in Mouse Intestinal T Lymphocytes

et al. 2001

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Substance P (SP), one of the most prevalent neuropeptides in gut, has been reported to have potent immune modulatory effects as a proinflammatory agent. The synthesis of SP and SP receptor expression in intraepithelial and lamina propria T lymphocytes of mouse intestine was investigated. Using RT-PCR analysis, it was demonstrated that SP receptor mRNA was exclusively expressed in intraepithelial and lamina propria T lymphocytes as well as their purified CD4+, CD8+ and CD4–CD8–CD3+ subsets. Messenger RNAs (mRNAs) for the two precursors of SP, β and γ-preprotachykinin-A, were also detected. These results were consistent in lymphocytes from both epithelium and lamina propria of small and large intestines, although the frequencies and/or intensities of mRNA expression varied. However, none of the findings could be repeated in splenic T lymphocytes. Activation of splenocytes with anti-CD3Ε-chain mAb and PMA did not induce expression of SP or its receptor mRNAs. Furthermore, both cytoplasmic and surface-bound SP was demonstrated in intestinal T lymphocytes using dual color immunocytochemistry and immunoflow cytometry. In vitro treatment with SP did not significantly change the size of the SP-immunoreactive T cell population, indicating the presence of SP receptor on intestinal T lymphocytes as well as in vivo binding of endogenously released SP. Our data suggest that SP production and SP receptor expression are distinctive for mouse intestinal mucosal immunity and that SP may act as a modulator of an ongoing controlled inflammation in normal gut, by acting through its specific receptor on T lymphocytes in an autocrine and/or paracrine pattern.

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“…The sections were then washed in three changes of PBS, mounted in glycerol:PBS (1:1) and examined under a Leitz Orthoplane photomicroscope equipped with epifluorescence optics. For details of the immunohistochemical procedures, see also Forsgren and Söderberg [1987]. In order to enhance specific immunofluorescence reactions, treatment with acid potassium permanganate was performed for 2 min before incubation in Triton X-100 [Hansson and Forsgren, 1995].…”

Section: Immunohistochemical Proceduresmentioning

confidence: 99%

Sympathetic and Sensory Innervations Are Heterogeneously Distributed in Relation to the Blood Vessels at the Extensor carpi radialis brevis Muscle Origin of Man

Ljung

Forsgren

Fridén³

1999

Cells Tissues Organs

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By using immunohistochemistry and antibodies to a general nerve marker, protein gene product (PGP) 9.5, the overall innervation at the extensor carpi radialis brevis (ECRB) muscle origin was investigated in patients with tennis elbow and in healthy controls. The autonomic innervation was studied by using antibodies to neuropeptide Y (NPY), tyrosine hydroxylase (TH), and vasoactive intestinal peptide (VIP). The sensory innervation was visualized by using antibodies to substance P and calcitonin gene-related peptide. PGP 9.5 immunoreactions were detected in association with small blood vessels and arteries and within nerve bundles. There was, however, heterogeneity in the perivascular nerve fiber distribution since some blood vessels exhibited a high degree of PGP 9.5 innervation and some negligible or no such innervation at all. There was marked TH/NPY innervation in the walls of a subpopulation of the arteries, basically no VIP-containing nerves, and sensory innervation restricted to the small blood vessels. These observations show that the ECRB muscle origin is supplied with heterogeneously distributed sympathetic and sensory innervations and, furthermore, that there appears to be an imbalance between the vasoconstrictor and vasodilator innervations along the vascular tree in this region.

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Immunohistochemical procedures for the demonstration of peptide- and tyrosine hydroxylase-containing nerve fibers in cryostat sections of unfixed rapidly frozen tissue stored for long periods of time

Cited by 54 publications

References 20 publications

Tyrosine hydroxylase positive nerves and mast cells in the porcine gallbladder

Tyrosine hydroxylase positive nerves and mast cells in the porcine gallbladder

Both Substance P and Its Receptor Are Expressed in Mouse Intestinal T Lymphocytes

Sympathetic and Sensory Innervations Are Heterogeneously Distributed in Relation to the Blood Vessels at the Extensor carpi radialis brevis Muscle Origin of Man

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