A new method for physical disector analyses of numbers and mean volumes of immunohistochemically labeled cells in paraffin sections

Hofmann, Isabel; Kemter, Elisabeth; Fiedler, Sonja; Theobalt, Natalie; Fonteyne, Lina Marie; Wolf, Eckhard; Wanke, Rüdiger; Blutke, Andreas

doi:10.1016/j.jneumeth.2021.109272

Cited by 5 publications

(5 citation statements)

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“…Pituitary glands were extracted as described previously 14 and weighed to the nearest milligram. The total volume of the PG (V (PG) ) was determined from its weight (m (PG) ) and its specific weight (γ (PG) = 0.89 g/cm 3 ) as V (PG) = m (PG) /γ (PG) 13,15 . Body weight and brain weight of the animals were also assessed in the same previous study 4 …”

Section: Methodsmentioning

confidence: 99%

“…For holistic proteome analysis, a sample of approximately 1 mm 3 was excised from the adenohypophysis of four GHR ‐KO and four control animals (2 males and 2 females each) and immediately frozen on dry ice. The remaining pituitary gland was processed as described previously 13–16 . After fixation of the tissue in neutrally buffered 4% formaldehyde solution for 24 h at room temperature, the pituitary gland was exhaustively sliced into approximately 1 mm thick, parallel slabs.…”

Section: Methodsmentioning

confidence: 99%

“…The total volume of the PG (V (PG) ) was determined from its weight (m (PG) ) and its specific weight (γ (PG) = 0.89 g/cm 3 ) as V (PG) = m (PG) /γ (PG) . 13,15 Body weight and brain weight of the animals were also assessed in the same previous study. 4 For holistic proteome analysis, a sample of approximately 1 mm 3 was excised from the adenohypophysis of four GHR-KO and four control animals (2 males and 2 females each) and immediately frozen on dry ice.…”

Section: Generation Of Pituitary Gland Tissue Samplesmentioning

confidence: 99%

“…We recently confirmed that GHR-KO pigs resemble the hallmark for GHRD, which are elevated GH and decreased IGF1 levels. 4 For an objective quantitative assessment of the effects of GHRD on relevant morphological characteristics of the porcine pituitary gland and somatotroph cells, established unbiased quantitative-stereological analyses were [13][14][15][16] applied, demonstrating that the volume proportions of the adenohypophysis in the pituitary gland, and of somatotroph cells in the adenohypophysis were not significantly altered in GHR-KO pigs.…”

Section: Ghrd Leads To Major Alterations Of the Adenohypophysis Prote...mentioning

confidence: 99%

“…The remaining pituitary gland was processed as described previously. [13][14][15][16] After fixation of the tissue in neutrally buffered 4% formaldehyde solution for 24 h at room temperature, the pituitary gland was exhaustively sliced into approximately 1 mm thick, parallel slabs.…”

Section: Generation Of Pituitary Gland Tissue Samplesmentioning

confidence: 99%

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Structural and proteomic repercussions of growth hormone receptor deficiency on the pituitary gland: Lessons from a translational pig model

Shashikadze

Franzmeier

Hofmann

et al. 2023

J Neuroendocrinology

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Growth hormone receptor deficiency (GHRD) results in low serum insulin‐like growth factor 1 (IGF1) and high, but non‐functional serum growth hormone (GH) levels in human Laron syndrome (LS) patients and animal models. This study investigated the quantitative histomorphological and molecular alterations associated with GHRD. Pituitary glands from 6 months old growth hormone receptor deficient (GHR‐KO) and control pigs were analyzed using a quantitative histomorphological approach in paraffin (9 GHR‐KO [5 males, 4 females] vs. 11 controls [5 males, 6 females]), ultrathin sections tissue sections (3 male GHR‐KO vs. 3 male controls) and label‐free proteomics (4 GHR‐KO vs. 4 control pigs [2 per sex]). GHR‐KO pigs displayed reduced body weights (60% reduction in comparison to controls; p < .0001) and decreased pituitary volumes (54% reduction in comparison to controls; p < .0001). The volume proportion of the adenohypophysis did not differ in GHR‐KO and control pituitaries (65% vs. 71%; p = .0506) and GHR‐KO adenohypophyses displayed a reduced absolute volume but an unaltered volume density of somatotrophs in comparison to controls (21% vs. 18%; p = .3164). In GHR‐KO pigs, somatotroph cells displayed a significantly reduced volume density of granules (23.5%) as compared to controls (67.7%; p < .0001). Holistic proteome analysis of adenohypophysis samples identified 4660 proteins, of which 592 were differentially abundant between the GHR‐KO and control groups. In GHR‐KO samples, the abundance of somatotropin precursor was decreased, whereas increased abundances of proteins involved in protein production, transport and endoplasmic reticulum (ER) stress were revealed. Increased protein production and secretion as well as significantly reduced proportion of GH‐storing granules in somatotroph cells of the adenohypophysis without an increase in volume density of somatotroph cells in the adenohypophysis could explain elevated serum GH levels in GHR‐KO pigs.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%