Limits...
Effect of E-cadherin expression on hormone production in rat anterior pituitary lactotrophs in vitro.

Kusumoto K, Kikuchi M, Fujiwara K, Horiguchi K, Kouki T, Kawanishi K, Yashiro T - Acta Histochem Cytochem (2010)

Bottom Line: Recent studies of the developing rat adenohypophysis found that primordial cells co-expressed E- and N-cadherins, but that hormone-producing cells lost E-cadherin and ultimately possessed only N-cadherin.The mean signal intensity of prolactin protein in rE-cad-IZ-transfected cells was approximately one fourth that in intact cells and in -IZ-transfected cells (P<0.01).These results suggest that the expression of E-cadherin does not affect prolactin mRNA transcription; rather, it reduces prolactin protein content, presumably by affecting trafficking of secretory granules.

View Article: PubMed Central - PubMed

Affiliation: Division of Histology and Cell Biology, Department of Anatomy, Jichi Medical University School of Medicine, Yakushiji 3311-1, Shimotsuke-shi, Tochigi 329-0498.

ABSTRACT
Cadherins are a family of transmembrane glycoproteins that mediate cell-to-cell adhesion. A change in cadherin type in cells, i.e., cadherin switching, induces changes in the character of the cell. Recent studies of the developing rat adenohypophysis found that primordial cells co-expressed E- and N-cadherins, but that hormone-producing cells lost E-cadherin and ultimately possessed only N-cadherin. In the present study, we examined the roles of cadherin switching in cytogenesis of anterior pituitary cells by observing prolactin mRNA and protein expression in lactotrophs that were transformed with an E-cadherin expression vector. In hormone-producing cells that were transfected with a pIRES2-ZsGreen1 plasmid with a full-length E-cadherin cDNA (rE-cad-IZ) insert in primary culture, we detected E- and N-cadherins on plasma membrane and E-cadherin in cytoplasm. In these rE-cad-IZ-transfected cells, in situ hybridization revealed prolactin mRNA signals that were at a level identical to that in control cells, while prolactin protein was barely detectable using immunocytochemistry. The mean signal intensity of prolactin protein in rE-cad-IZ-transfected cells was approximately one fourth that in intact cells and in -IZ-transfected cells (P<0.01). These results suggest that the expression of E-cadherin does not affect prolactin mRNA transcription; rather, it reduces prolactin protein content, presumably by affecting trafficking of secretory granules.

No MeSH data available.


Related in: MedlinePlus

In situ hybridization and immunocytochemistry of transfected anterior pituitary cells. Anterior pituitary cells in primary culture were transfected with -IZ (A, C, E and G, in the identical field) and with rE-cad-IZ (B, D, F and H, in the identical field). A and B: Differential interference contrast images of transformed cells. C and D: Merged images of immunoreaction of prolactin protein (red), in situ hybridization signal for prolactin mRNA (light blue), and fluorescence from ZsGreen1 (C and D). E and F: Immunoreaction of prolactin. G and H: In situ hybridization signals for prolactin mRNA. Arrows and arrowheads show cells transfected with rE-cad-IZ and -IZ, respectively. Bar=5 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2875860&req=5

Figure 2: In situ hybridization and immunocytochemistry of transfected anterior pituitary cells. Anterior pituitary cells in primary culture were transfected with -IZ (A, C, E and G, in the identical field) and with rE-cad-IZ (B, D, F and H, in the identical field). A and B: Differential interference contrast images of transformed cells. C and D: Merged images of immunoreaction of prolactin protein (red), in situ hybridization signal for prolactin mRNA (light blue), and fluorescence from ZsGreen1 (C and D). E and F: Immunoreaction of prolactin. G and H: In situ hybridization signals for prolactin mRNA. Arrows and arrowheads show cells transfected with rE-cad-IZ and -IZ, respectively. Bar=5 µm.

Mentions: We detected prolactin immunoreactivity and mRNA signals in primary culture by means of immunocytochemical and in situ hybridization techniques. In intact cells, obvious prolactin immunoreactivity (Fig. 2C, 2D, 2E, and 2F) and mRNA signals (Fig. 2C, 2D, 2G and 2H) were obvious. In -IZ-transfected cells (indicated by the arrowheads in Fig. 2), prolactin immunoreactivity and mRNA signals (Fig. 2C, 2E and 2G) were identical to those detected in intact cells. In contrast, in rE-cad-IZ-transfected cells (indicated by the arrows in Fig. 2), prolactin mRNA signals (Fig. 2D and 2H) were detected, but prolactin immunoreactivity (Fig. 2C and 2F) was barely detectable.


Effect of E-cadherin expression on hormone production in rat anterior pituitary lactotrophs in vitro.

Kusumoto K, Kikuchi M, Fujiwara K, Horiguchi K, Kouki T, Kawanishi K, Yashiro T - Acta Histochem Cytochem (2010)

In situ hybridization and immunocytochemistry of transfected anterior pituitary cells. Anterior pituitary cells in primary culture were transfected with -IZ (A, C, E and G, in the identical field) and with rE-cad-IZ (B, D, F and H, in the identical field). A and B: Differential interference contrast images of transformed cells. C and D: Merged images of immunoreaction of prolactin protein (red), in situ hybridization signal for prolactin mRNA (light blue), and fluorescence from ZsGreen1 (C and D). E and F: Immunoreaction of prolactin. G and H: In situ hybridization signals for prolactin mRNA. Arrows and arrowheads show cells transfected with rE-cad-IZ and -IZ, respectively. Bar=5 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2875860&req=5

Figure 2: In situ hybridization and immunocytochemistry of transfected anterior pituitary cells. Anterior pituitary cells in primary culture were transfected with -IZ (A, C, E and G, in the identical field) and with rE-cad-IZ (B, D, F and H, in the identical field). A and B: Differential interference contrast images of transformed cells. C and D: Merged images of immunoreaction of prolactin protein (red), in situ hybridization signal for prolactin mRNA (light blue), and fluorescence from ZsGreen1 (C and D). E and F: Immunoreaction of prolactin. G and H: In situ hybridization signals for prolactin mRNA. Arrows and arrowheads show cells transfected with rE-cad-IZ and -IZ, respectively. Bar=5 µm.
Mentions: We detected prolactin immunoreactivity and mRNA signals in primary culture by means of immunocytochemical and in situ hybridization techniques. In intact cells, obvious prolactin immunoreactivity (Fig. 2C, 2D, 2E, and 2F) and mRNA signals (Fig. 2C, 2D, 2G and 2H) were obvious. In -IZ-transfected cells (indicated by the arrowheads in Fig. 2), prolactin immunoreactivity and mRNA signals (Fig. 2C, 2E and 2G) were identical to those detected in intact cells. In contrast, in rE-cad-IZ-transfected cells (indicated by the arrows in Fig. 2), prolactin mRNA signals (Fig. 2D and 2H) were detected, but prolactin immunoreactivity (Fig. 2C and 2F) was barely detectable.

Bottom Line: Recent studies of the developing rat adenohypophysis found that primordial cells co-expressed E- and N-cadherins, but that hormone-producing cells lost E-cadherin and ultimately possessed only N-cadherin.The mean signal intensity of prolactin protein in rE-cad-IZ-transfected cells was approximately one fourth that in intact cells and in -IZ-transfected cells (P<0.01).These results suggest that the expression of E-cadherin does not affect prolactin mRNA transcription; rather, it reduces prolactin protein content, presumably by affecting trafficking of secretory granules.

View Article: PubMed Central - PubMed

Affiliation: Division of Histology and Cell Biology, Department of Anatomy, Jichi Medical University School of Medicine, Yakushiji 3311-1, Shimotsuke-shi, Tochigi 329-0498.

ABSTRACT
Cadherins are a family of transmembrane glycoproteins that mediate cell-to-cell adhesion. A change in cadherin type in cells, i.e., cadherin switching, induces changes in the character of the cell. Recent studies of the developing rat adenohypophysis found that primordial cells co-expressed E- and N-cadherins, but that hormone-producing cells lost E-cadherin and ultimately possessed only N-cadherin. In the present study, we examined the roles of cadherin switching in cytogenesis of anterior pituitary cells by observing prolactin mRNA and protein expression in lactotrophs that were transformed with an E-cadherin expression vector. In hormone-producing cells that were transfected with a pIRES2-ZsGreen1 plasmid with a full-length E-cadherin cDNA (rE-cad-IZ) insert in primary culture, we detected E- and N-cadherins on plasma membrane and E-cadherin in cytoplasm. In these rE-cad-IZ-transfected cells, in situ hybridization revealed prolactin mRNA signals that were at a level identical to that in control cells, while prolactin protein was barely detectable using immunocytochemistry. The mean signal intensity of prolactin protein in rE-cad-IZ-transfected cells was approximately one fourth that in intact cells and in -IZ-transfected cells (P<0.01). These results suggest that the expression of E-cadherin does not affect prolactin mRNA transcription; rather, it reduces prolactin protein content, presumably by affecting trafficking of secretory granules.

No MeSH data available.


Related in: MedlinePlus