Cellular localization of nerve growth factor synthesis by in situ hybridization.

Nerve growth factor (NGF) in mouse submandibular glands (SGs) is generated from a 35-kD precursor by proteolytic enzymes that have yet to be identified. Prohormone convertases (PCs) cleave the NGF precursor in vitro, and in this study we questioned whether PCs could process salivary NGF in vivo. mRNA coding for PC2 (but not PC1) was detected on Northern blots of SG mRNA and also by in situ hybridization within parasympathetic neurons of intralobular ganglia. Northern blot and in situ hybridization analyses also detect mRNA coding for furin. In SGs of male mice, furin mRNA levels are high at birth and remain high throughout development. In glands from female mice, levels decline during postnatal development and are lower in adults than in newborns. Immunocytochemistry detects furin immunoreactivity in pro-acinar and ductal cells of glands from newborn and pubescent mice. In glands of adults, furin immunoreactivity is detectable in acinar cells but highest levels are present in NGF-containing granular convoluted tubule cells. These data, taken together with those from previous studies, suggest that furin is a candidate processing enzyme for NGF in mouse submandibular glands.

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Nerve growth factor mRNA in brain: Localization by in situ hybridization

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(86)80569-1 ◽

1986 ◽

Vol 138 (2) ◽

pp. 813-818 ◽

Cited By ~ 89

Author(s):

Paul D. Rennert ◽

Gerhard Heinrich

Keyword(s):

In Situ Hybridization ◽

Nerve Growth Factor ◽

Growth Factor ◽

Nerve Growth ◽

Brain Localization

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In Situ Hybridization for Neurofilament and GAP-43 mRNA in Primary Sensory Neurons in Conjunction with Nerve Growth Factor Receptor Radioautography

Methods in Neurosciences - Gene Expression in Neural Tissues ◽

10.1016/b978-0-12-185267-2.50023-4 ◽

1992 ◽

pp. 274-287 ◽

Cited By ~ 4

Author(s):

W. Tetzlaff ◽

V.M.K. Verge ◽

M.A. Bisby ◽

P.M. Richardson

Keyword(s):

In Situ Hybridization ◽

Nerve Growth Factor ◽

Growth Factor ◽

Sensory Neurons ◽

Growth Factor Receptor ◽

Nerve Growth Factor Receptor ◽

Primary Sensory Neurons ◽

Nerve Growth

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Detection of nerve growth factor mRNA in rodent salivary glands with digoxigenin- and 33P-labeled oligonucleotides: effects of castration and sympathectomy.

Journal of Histochemistry & Cytochemistry ◽

10.1177/41.5.8468451 ◽

1993 ◽

Vol 41 (5) ◽

pp. 703-708 ◽

Cited By ~ 12

Author(s):

C Humpel ◽

E Lindqvist ◽

L Olson

Keyword(s):

In Situ Hybridization ◽

Nerve Growth Factor ◽

Growth Factor ◽

Salivary Glands ◽

Submandibular Gland ◽

Male Mouse ◽

Male Mice ◽

Nerve Growth ◽

Submandibular Glands

Nerve growth factor (NGF) is a protein highly expressed in the male mouse submandibular gland. We have applied a non-radioactive in situ hybridization method using digoxigenin-labeled NGF oligonucleotides, and have found the highest amounts of NGF mRNA in the secretory striated ducts of the male mouse submandibular gland. Scattered strongly positive cells were found in male mouse sublingual glands. Weakly labeled cells were seen in female mouse and in male rat submandibular gland striated duct cells. Using 33P as an alternative to 32P and 35S, we demonstrated a 1.3 KB NGF mRNA in salivary glands of male mice by Northern blot hybridization. Using 33P we detected NGF mRNA in male mouse submandibular glands by in situ hybridization but with a signal that, compared with the non-radioactive method, had a very low resolution. Castration of male mice almost abolished both the 1.3 KB NGF mRNA seen with Northern blots and the NGF mRNA labeling in submandibular glands 4 weeks after the operation, whereas levels were increased 6 hr and 2 days after sympathectomy. We conclude that hybridization with digoxigenin-labeled NGF oligonucleotides is a good tool to study the expression and regulation of NGF mRNA in male mouse submandibular glands.

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