scholarly journals Immunocytochemical localization of cathepsin B in rat anterior pituitary endocrine cells, with special reference to its co-localization with renin and prorenin in gonadotrophs.

1991 ◽  
Vol 39 (9) ◽  
pp. 1199-1205 ◽  
Author(s):  
Y Uchiyama ◽  
M Nakajima ◽  
T Watanabe ◽  
S Waguri ◽  
N Sato ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Luteinizing Hormone ◽  
Light Microscopy ◽  
Cathepsin B ◽  
Anterior Pituitary ◽  
Endocrine Cells ◽  
Secretory Granules ◽  
Juxtaglomerular Cells ◽  
Immunocytochemical Localization ◽  
Double Immunostaining

We examined by immunocytochemistry the localization of cathepsin B in endocrine cells of rat anterior pituitary lobe, using a monospecific antibody to cathepsin B. By light microscopy, granular immunodeposits for cathepsin B were detected in most endocrine cells of anterior pituitary lobe. Cells immunoreactive for luteinizing hormone (LH) were diffusely immunostained by anti-cathepsin B. By electron microscopy, immunogold particles for cathepsin B were localized in lysosomes of thyrotrophs, somatotrophs, and mammotrophs. In mammotrophs, immunogold particles for cathepsin B were also detected in crinophagic bodies. Double immunostaining co-localized immunogold particles for LH and cathepsin B in secretory granules of gonadotrophs. Immunocytochemistry was also applied to demonstrate localization of renin and prorenin in LH-producing gonadotrophs; immunogold particles for renin were co-localized with those for LH, cathepsin B, or prorenin in their secretory granules. Immunogold particles for prorenin were also co-localized with those for LH or cathepsin B in secretory granules, but prorenin-positive granules appeared less frequently than renin-positive granules. These results suggest that cathepsin B not only plays a role in the protein degradation in lysosomes of anterior pituitary endocrine cells but also participates in the activation of renin in gonadotrophs, as has been demonstrated in secretory granules of juxtaglomerular cells.

scholarly journals Immunocytochemical localization of prorenin, renin, and cathepsins B, H, and L in juxtaglomerular cells of rat kidney.

1989 ◽  
Vol 37 (11) ◽  
pp. 1689-1697 ◽  
Author(s):  
H Matsuba ◽  
T Watanabe ◽  
M Watanabe ◽  
Y Ishii ◽  
S Waguri ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cathepsin B ◽  
Rat Kidney ◽  
Renal Tissue ◽  
Juxtaglomerular Cells ◽  
Perinuclear Region ◽  
Immunocytochemical Localization ◽  
Double Immunostaining ◽  
Thin Sections ◽  
Anti Body

To examine the correlation of localization of prorenin, renin, and cathepsins B, H, and L, immunocytochemistry was applied to rat renal tissue, using a sequence-specific anti-body (anti-prorenin) that recognizes the COOH terminus of the rat renin prosegment. In serial semi-thin sections, immunodeposits for prorenin, renin, and cathepsins B, H, and L were localized in the same juxtaglomerular (JG) cells. Immunodeposits for renin were detected throughout the cytoplasm of the cells, whereas those for prorenin were detected in the perinuclear region. Immunoreactivity for cathepsin B was stronger than that for cathepsins H and L. By electron microscopy, prorenin was localized in small (immature) granules but not in large mature granules, whereas renin was localized mainly in mature granules. In serial thin sections, prorenin, renin, and cathepsin B were colocalized in the same immature granules containing heterogeneously dense material (intermediate granules). By double immunostaining, co-localization of renin with cathepsins B, H, or L was demonstrated in mature granules. The results suggest the possibility that processing of prorenin to renin occurs in immature granules of rat JG cells, and cathepsin B detected in JG cells may be a major candidate for the maturation of renin.

Dopamine enters lactotrophs and reaches their secretory granules

1985 ◽  
Vol 104 (1) ◽  
pp. 23-NP ◽  
Author(s):  
M. G. P. Gallardo ◽  
M. Bilinski ◽  
S. R. Chiocchio ◽  
J. H. Tramezzani
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Anterior Pituitary ◽  
Secretory Granules ◽  
Pituitary Cells ◽  
Histochemical Reaction ◽  
Anterior Pituitary Cells ◽  
Biochemical Assays ◽  
Pituitary Lactotroph

ABSTRACT The presence of dopamine in the lactotroph cell, as well as in isolated prolactin secretory granules, was demonstrated by means of an histochemical reaction for electron microscopy. Biochemical assays further confirmed the presence of dopamine in the secretory granules. Autoradiographic preparations examined by light microscopy showed dopamine internalization in dispersed anterior pituitary cells. Isolated anterior pituitary lactotroph cells incorporated more [3H]dopamine than a fraction containing other anterior pituitary cells. J. Endocr. (1985) 104, 23–28

scholarly journals ISOLATION AND BIOLOGICAL PROPERTIES OF SECRETORY GRANULES FROM RAT ANTERIOR PITUITARY GLANDS

1969 ◽  
Vol 43 (3) ◽  
pp. 564-574 ◽  
Author(s):  
Allen Costoff ◽  
W. H. McShan
Keyword(s):  
Electron Microscopy ◽  
Anterior Pituitary ◽  
Close Agreement ◽  
Secretory Granules ◽  
Cell Types ◽  
Pituitary Hormones ◽  
Biological Properties ◽  
Single Fraction ◽  
Rat Pituitary ◽  
Pituitary Glands

A method is described for the isolation of secretory granules from rat anterior pituitary glands. The method consists of differential and isopycnic gradient centrifugations, followed by filtration of the zones containing granules on Nuclepore filters to remove mitochondria. Highly purified granules were obtained as indicated by electron microscopy. Major parts of the thyrotropin (TSH) and adrenocorticotropin (ACTH) were recovered in a single fraction of granules as were follicle-stimulating (FSH) and luteinizing (LH) hormones. The somatotropin (STH) and prolactin (LTH) were recovered in separate granule fractions. The major parts of the six different hormones were associated with their respective granule fractions as shown by bioassays specific for each of the hormones. The diameters of granules in sections of intact rat pituitary glands and in isolated pellets were measured, and the means and ranges were in close agreement. These results contribute to the identification of the cell types which produce the different pituitary hormones.

scholarly journals Immunocytochemical localization of atrial natriuretic factor in the kidney, adrenal medulla, pituitary, and atrium of rat.

1985 ◽  
Vol 33 (8) ◽  
pp. 828-832 ◽  
Author(s):  
J C McKenzie ◽  
I Tanaka ◽  
K S Misono ◽  
T Inagami
Keyword(s):  
Adrenal Medulla ◽  
Anterior Pituitary ◽  
Secretory Granules ◽  
Immunocytochemical Localization ◽  
Atrial Myocytes ◽  
Collecting Tubules ◽  
Sodium Regulation ◽  
Sites Of Action ◽  
A Site ◽  
Atrial Natriuretic

Mammalian atria have previously been shown to produce a variety of peptides with natriuretic and vasorelaxant activities. Certain of these atrial natriuretic factors (ANF) have been localized immunocytochemically in secretory granules of atrial myocytes. However, the precise sites of action and extra-atrial synthesis or accumulation of ANF have not been identified immunocytochemically. In the present study, immunoreactive ANF was detected in rat atrial myocytes, intercalated cells of the renal collecting ducts, adrenal medullary chromaffin cells, and gonadotrophs of the anterior pituitary using an antibody against synthetic rat ANF-IV (H2N-Arg-Ser-Ser-Cys-Phe-Gly- Gly-Arg-Ile-Asp-Arg-Ile-Gly-Ala-Gln-Ser-Gly-Leu-Gly-Cys-Asn-Ser-Phe- Arg-Try-COOH). The localization of ANF in specialized cells of the renal collecting tubules and ducts supports suggestions that these structures may be a site of natriuretic action of ANF. In addition, immunocytochemical localization of ANF in the rat adrenal medulla and anterior pituitary suggests the existence of alternate sites of action and/or synthesis. We believe these findings are important for a more complete understanding of the role of ANF in fluid and sodium regulation and of the participation of ANF in the development of sodium-dependent hypertension.

scholarly journals Immunocytochemical localization of cathepsins B, H, and their endogenous inhibitor, cystatin beta, in islet endocrine cells of rat pancreas.

1988 ◽  
Vol 36 (7) ◽  
pp. 783-791 ◽  
Author(s):  
M Watanabe ◽  
T Watanabe ◽  
Y Ishii ◽  
H Matsuba ◽  
S Kimura ◽  
...  
Keyword(s):  
Endocrine Cells ◽  
Islet Cells ◽  
Secretory Granules ◽  
Double Immunostaining ◽  
Thin Sections ◽  
Cathepsin H ◽  
Immunocytochemical Techniques ◽  
The Difference ◽  
Immunocytochemical Reaction ◽  
Strong Immunoreactivity

To determine the characteristics of lysosomes in rat islet endocrine cells, we examined the precise localization of cathepsins B, H, and L and their specific inhibitors, cystatins alpha and beta, using immunocytochemical techniques. By use of serial semi-thin sections, we detected immunoreactivity for cathepsin B in insulin-, glucagon-, somatostatin-, and pancreatic polypeptide-positive (PP) cells. Strong immunoreactivity for cathepsin H was seen in A-cells and weak immunoreactivity in PP cells, but none in others. Immunodeposits for cystatin beta were demonstrated in B-cells. Brief dipping of thin sections in 1% sodium methoxide before the following immunocytochemical reaction enhanced specific deposits of immunogold particles on the target organelles. Use of a double-immunostaining technique showed co-localization of insulin with cystatin beta in many secretory granules. This suggests that cystatin beta may regulate converting enzymes participating in the maturation process of insulin. By use of an immunogold technique, heterogeneous localization of cathepsins B and H in lysosomes was also found among islet cells at the light microscopic level. This may be due to the difference in peptides degraded in lysosomes among the cells.

scholarly journals The major tyrosine-sulfated protein of the bovine anterior pituitary is a secretory protein present in gonadotrophs, thyrotrophs, mammotrophs, and corticotrophs.

1985 ◽  
Vol 100 (3) ◽  
pp. 928-937 ◽  
Author(s):  
P Rosa ◽  
G Fumagalli ◽  
A Zanini ◽  
W B Huttner
Keyword(s):  
Anterior Pituitary ◽  
Endocrine Cells ◽  
Secretory Granules ◽  
Secretory Protein ◽  
Tyrosine Sulfation ◽  
Biochemical Studies ◽  
Immunofluorescence Studies ◽  
The Matrix ◽  
Corticotrophic Cells ◽  
Secretory Tissue

The anterior pituitary is a complex secretory tissue known to contain several sulfated macromolecules. In the present study, we identified the major tyrosine-sulfated protein of the bovine anterior pituitary and investigated its cellular and subcellular localization. This protein consisted of two tyrosine-sulfated polypeptides of molecular weight 86,000 and 84,000 that were highly homologous to each other. In agreement with previous biochemical studies, the tyrosine-sulfated protein of Mr 86,000/84,000 was found to be secretory, as it was observed in the matrix of secretory granules by immunoelectron microscopy. Immunofluorescence studies indicated that the tyrosine-sulfated, secretory protein of Mr 86,000/84,000, referred to as TSP 86/84, was present in all endocrine cells except for some somatotrophic cells. Higher levels of immunoreactivity for TSP 86/84 were observed in gonadotrophic and thyrotrophic than in mammotrophic and corticotrophic cells. This appeared to result from the occurrence of TSP 86/84 in all secretory granules of the former cells and in only some secretory granules of the latter cells. We discuss the possibility that TSP 86/84 may have a role in the packaging of several distinct peptides hormones into secretory granules. One, though not the only, possible function of tyrosine sulfation may concern the sorting of this protein in the Golgi complex.

Immunocytochemical localization of lactase in the brush border, Golgi complex and endocytotic granules of pig jejunal enterocytes

1994 ◽  
Vol 52 ◽  
pp. 292-293
Author(s):  
Julian P. Heath ◽  
Rhonda G. Harrell ◽  
László G. Kömüves
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Golgi Complex ◽  
Brush Border ◽  
Immunogold Electron Microscopy ◽  
Immunocytochemical Localization ◽  
Developmentally Regulated ◽  
Neonatal Piglets ◽  
Brush Border Enzyme ◽  
Crypt Cells

We have used immunofluorescence light microscopy (IFLM) and immunogold electron microscopy (IEM) to study the distribution of lactase phlorizin hydrolase, a developmentally regulated brush border enzyme, in the enterocytes of suckling neonatal piglets. Tissues were collected from the jejunum and fixed in 4% paraformaldehyde. Cryosections were incubated in mouse monoclonal anti-pig lactase antibody (PBB 3/7/3/2, from Dr. A. Quaroni, Cornell Univ.) followed by either FITC- or 10 nm gold-conjugated anti-mouse IgG. IFLM and IEM images were digitally acquired from video and processed.Jejunal villus enterocytes of non-suckled piglets possess an apical endocytic complex and a sub-nuclear Golgi complex (Fig. 1). In the jejunal crypts, differentiating enterocytes possess an immature brush border and a supra-nuclear Golgi complex (Fig. 2). In suckled animals, absorption of colostral proteins by villus enterocytes results in the formation of dense apical and basal granules (Fig. 3); crypt cells do not accumulate granules. Many of the granules are juxtaposed to the Golgi complex (Fig. 4).

scholarly journals Coexistence of renin and cathepsin B in secretory granules of granular duct cells in male mouse submandibular gland.

1993 ◽  
Vol 41 (3) ◽  
pp. 433-438 ◽  
Author(s):  
K Sano ◽  
S Waguri ◽  
N Sato ◽  
E Kominami ◽  
Y Uchiyama
Keyword(s):  
Submandibular Gland ◽  
Golgi Complex ◽  
Cathepsin B ◽  
Cysteine Proteinase ◽  
Secretory Granules ◽  
Interstitial Cells ◽  
Double Immunostaining ◽  
Duct Cells ◽  
Cathepsin H ◽  
Rat Pituitary

Cathepsin B, a representative lysosomal cysteine proteinase, has been demonstrated to coexist with renin in secretary granules of rat pituitary LH/FSH cells and renal juxtaglomerular cells. We investigated immunocytochemically the localization of cathepsins B, H, and L in the submandibular gland of male mice, in which active renin is also produced. By light microscopy, granular immunodeposits for cathepsin B were detected in epithelial cells of the gland, particularly in granular duct cells and interstitial cells. Immunoreactivity for cathepsins H and L was mainly found in interstitial cells, although that for cathepsin H was weakly seen in acinar cells. By electron microscopy, immunogold particles indicating cathepsin B intensely labeled small granules near the Golgi complex of granular duct cells and weakly labeled large secretory granules, whereas those showing renin labeled both granules. Double immunostaining co-localized immunogold particles showing renin and cathepsin B in small perinuclear granules near the Golgi complex. Some immunopositive granules seemed to be closely associated with the Golgi elements. These results indicate that the co-localization of renin and cathepsin B is also seen in secretory granules of granular duct cells in the mouse submandibular gland, as seen in rat juxtaglomerular and LH/FSH cells. This suggests that cathepsin B is one of the possible candidates for the renin-processing enzyme.

scholarly journals Cysteine proteinases in rat parathyroid cells with special reference to their correlation with parathyroid hormone (PTH) in storage granules.

1993 ◽  
Vol 41 (2) ◽  
pp. 273-282 ◽  
Author(s):  
Y Hashizume ◽  
S Waguri ◽  
T Watanabe ◽  
E Kominami ◽  
Y Uchiyama
Keyword(s):  
Electron Microscopy ◽  
Parathyroid Hormone ◽  
Special Reference ◽  
Secretory Granules ◽  
Cysteine Proteinases ◽  
Double Immunostaining ◽  
Thin Sections ◽  
Storage Granules ◽  
Golgi Network ◽  
And Storage

To further understand the roles of storage granules in parathyroid cells, we examined by immunocytochemistry the localization of cathepsins B and H and of PTH in rat parathyroid gland. In semi-thin sections, small and large granular immunodeposits for cathepsins B and H appeared in the cells, whereas those for PTH were detected throughout the cells, especially in perinuclear regions. By electron microscopy, immunogold particles indicating cathepsins B and H labeled lysosomes and storage granules, whereas those showing PTH were localized in storage granules, small secretory granules, and the trans-Golgi network. Small vesicles labeled by immunogold particles showing these proteinases often appeared close to the storage granules. By double immunostaining, immunogold particles indicating these proteinases were co-localized with those for PTH in storage granules. By EDTA treatment, immunoreactivity for cathepsins B and H and for PTH was notably reduced in the cells, but immunoreactivity for the proteinases was still seen in lysosomes. These results suggest that storage granules in the rat parathyroid cells fuse with small vesicles containing cathepsins B and H, which may participate in regulating the intracellular PTH levels by degrading PTH in the granules.

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